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Acacia modesta (AM) and Opuntia monocantha (OM) are distributed in Pakistan, Afghanistan and India. Both of these plants have different pharmacological properties. This study was designed to evaluate anticancer potential of Acacia modesta (AM) and Opuntia monocantha (OM). Liver cancer cell line HepG2 was used for assessment of anticancer activity. For the evaluation of anti-proliferative effects, cell viability and cell death in all groups of cells were evaluated via MTT, crystal violet and trypan blue assays. For the evaluation of apoptosis ELISA of p53 performed. Furthermore, LDH assay to find out the ability of malignant cells to metabolize pyruvate to lactate and antioxidant enzymes activity (GSH, CAT and SOD) at the end HPLC was performed to find active compound of AM and OM. Cytotoxicity (MTT), Viability assays (trypan blue, crystal viability, MUSE analysis) showed more dead, less live cells in plant treated groups with increase of concentration. Scratch assay for the anti-migratory effect of these plants showed treated groups have not ability to heal scratch/wound. ELISA of p53 for cellular apoptosis showed more release of p53 in treated groups. Antioxidant assay via glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) showed less anti-oxidative potential in treated cancer groups. LDH assay showed more lactate dehydrogenase release in treated groups compared with untreated. HPLC analysis showed the presence of phytochemicals such as steroids, alkaloids, phenols, flavonoids, saponins, tannins, anthraquinone and amino acids in AM and OM plant extracts. Based on all these findings, it can be concluded that ethanolic extracts of Acacia modesta and Opuntia monocantha have promising anti-cancer potential. Resumo Acacia modesta (AM) e Opuntia monocantha (OM) estão distribuídas no Paquistão, Afeganistão e Índia. Ambas as plantas têm propriedades farmacológicas diferentes. Este estudo foi desenhado para avaliar o potencial anticancerígeno de Acacia modesta (AM) e Opuntia monocantha (OM). A linha celular de câncer de fígado HepG2 foi usada para avaliação da atividade anticâncer. Para a avaliação dos efeitos antiproliferativos, a viabilidade celular e a morte celular em todos os grupos de células foram avaliadas através dos ensaios de MTT, cristal violeta e azul de tripano. Para a avaliação da apoptose foi realizado ELISA de p53. Além disso, o ensaio de LDH para descobrir a capacidade das células malignas de metabolizar o piruvato em lactato e a atividade das enzimas antioxidantes (GSH, CAT e SOD) no final da HPLC foi realizado para encontrar o composto ativo de AM e OM. Citotoxicidade (MTT), ensaios de viabilidade (azul de tripano, viabilidade de cristal, análise MUSE) mostraram mais células mortas e menos células vivas nos grupos tratados com plantas com aumento da concentração. O ensaio de arranhão para o efeito anti-migratório dessas plantas mostrou que os grupos tratados não têm capacidade de cicatrizar arranhão/ferida. ELISA de p53 para apoptose celular mostrou maior liberação de p53 nos grupos tratados. Ensaio antioxidante via glutationa (GSH), superóxido dismutase (SOD), catalase (CAT) mostrou menor potencial antioxidante nos grupos de câncer tratados. O ensaio de LDH mostrou mais liberação de lactato desidrogenase nos grupos tratados em comparação com os não tratados. A análise de HPLC mostrou a presença de fitoquímicos como esteróides, alcalóides, fenóis, flavonóides, saponinas, taninos, antraquinona e aminoácidos nos extratos vegetais AM e OM. Com base em todos esses achados, pode-se concluir que os extratos etanólicos de Acacia modesta e Opuntia monocantha apresentam potencial anticancerígeno promissor.

Quantum dots (QDs) have different properties: high electron density, magnetic moment, phosphorescence, photoluminescence (fluorescence), and strong optical absorption. The layer or ligands on the QDs surface has a vital role because they allow the stabilization and practical uses on different matrixes. Ligand exchange is a commonly carried out methodology to incorporate functional groups that alter the solubility, introduce electron transfer partners, integrate biological receptors, or improve the properties of the QDs surface. CdSSe QDs were synthesized using a microwave system using thioglycolic acid (TGA) as a sulfur source and cover agent. The TGA ligand was interchanged with cysteine (Cys), glutamic acid (GA), glutathione (GTO), glutaraldehyde (GLT), and lysine (Lys). The viability and response of the CYP1A1, CYP1A2, and CYP3A4 isoenzymes were directly measured in HEP-G2 cells after exposure to CdSSe-TGA, CdSSe-Cys, CdSSe-GA, CdSSe-GTO, CdSSe-GLT, and CdSSe-Lys. CdSSe and CdSSe-GTO (10 mg/L) decrease viability by around 65%. The response of the cytochrome isoenzymes is based on the organic ligand on the surface of the CdSSe QDs. Changes in CYP 1A1 could be related to carcinogenic xenobiotics. Fluorescence microscopy shows CdSSe QDs on and inside HEPG2 cells. The results confirm that apoptosis and necrosis are the principal mechanisms of decreased viability.

Drug induced liver injury (DILI) is one of the major reasons of drug withdrawal during the different phases of drug development. The later in the drug development a drug is discovered to be toxic, the higher the economical as well as the ethical impact will be. In vitro models for early detection of drug liver toxicity are under constant development, however to date a superior model of the liver is still lacking. Ideally, a highly reliable model should be established to maintain the different hepatic cell functionalities to the greatest extent possible, during a period of time long enough to allow for tracking of the toxicity of compounds. In the case of DILI, toxicity can appear even after months of exposure. To reach this goal, an in vitro model should be developed that mimics the in vivo liver environment, function and response to external stimuli. The different approaches for the development of liver models currently used in the field of tissue engineering will be described in this review. Combining different technologies, leading to optimal materials, cells and 3D-constructs will ultimately lead to an ideal superior model that fully recapitulates the liver.

The alcohol extracts of Chinese bayberry (Myrica rubra) branches (MRBE) are rich in flavonoids which have a variety of medicinal benefits, but their effects on human HepG2 were unknown. In this study, the effects of MRBE on HepG2 cell growth and its potential for inhibiting cancer were explored. The results displayed that MRBE inhibited HepG2 proliferation both by arresting cells in S phase and promoting apoptosis. Quantitative reverse-transcription PCR (qRT-PCR), western blotting, and immunofluorescence showed that MRBE induced S-phase arrest by upregulating p21, which in turn downregulated cyclin and cyclin-dependent kinase messenger RNA (mRNA) and protein. Apoptosis was induced by blocking the expression of BCL-2 and suppression of the Raf/ERK1 signaling pathways. These results indicated that MRBE may have the potential for treatment of human liver cancer, highlighting novel approaches for developing new pharmacological tools for the treatment of this deadly type cancer. Meanwhile, it provides a new direction for the medicinal added values of Chinese bayberry, which helped to broaden the diversified development of its industry chain.

Long non-coding RNAs (lncRNAs) are regulated in cancer cells, including lncRNA MEG3, which is downregulated in Hepatocellular Carcinoma (HCC). In addition, hepatitis C virus (HCV) core proteins are known to dysregulate important cellular pathways that are linked to HCC development. In this study, we were interested in evaluating the overexpression of lncRNA MEG3, either alone or in combination with two forms of HCV core protein (C173 and C191) in HepG2 cells. Cell viability was assessed by MTT assay. Transcripts’ levels of key genes known to be regulated in HCC, such as p53, DNMT1, miRNA152, TGF-b, and BCL-2, were measured by qRT-PCR. Protein expression levels of caspase-3 and MKI67 were determined by immunocytochemistry and apoptosis assays. The co-expression of lncRNA MEG3 and C191 resulted in a marked increase and accumulation of dead cells and a reduction in cell viability. In addition, a marked increase in the expression of tumor suppressor genes (p53 and miRNA152), as well as a marked decrease in the expression of oncogenes (DNMT1, BCL2, and TGF-b), were detected. Moreover, apoptosis assay results revealed a significant increase in total apoptosis (early and late). Finally, immunocytochemistry results detected a significant increase in apoptotic marker caspase-3 and a decrease in tumor marker MKI67. In this study, transgene expression of C191 and lncRNA MEG3 showed induction in apoptosis in HepG2 cells greater than the expression of each one alone. These results suggest potential anticancer characteristics.

Liver cancer is a prevalent form of cancer worldwide. While research has shown that increasing sphingomyelin (SM) hydrolysis by activating the cell surface membrane-associated neutral sphingomyelinase 2 (nSMase2) can control cell proliferation and apoptosis, the role of total glutathione depletion in inducing tumor cell apoptosis via nSMase2 activation is still under investigation. Conversely, glutathione-mediated inhibition of reactive oxygen species (ROS) accumulation is necessary for the enzymatic activity of nSMase1 and nSMase3, increased ceramide levels, and cell apoptosis. This study evaluated the effects of depleting total glutathione in HepG2 cells using buthionine sulfoximine (BSO). The study assessed nSMases RNA levels and activities, intracellular ceramide levels, and cell proliferation using RT-qPCR, Amplex red neutral sphingomyelinase fluorescence assay, and colorimetric assays, respectively. The results indicated a lack of nSMase2 mRNA expression in treated and untreated HepG2 cells. Depletion of total glutathione resulted in a significant increase in mRNA levels but a dramatic reduction in the enzymatic activity of nSMase1 and nSMase3, a rise in ROS levels, a decrease in intracellular levels of ceramide, and an increase in cell proliferation. These findings suggest that total glutathione depletion may exacerbate liver cancer (HCC) and not support using total glutathione-depleting agents in HCC management. It is important to note that these results are limited to HepG2 cells, and further studies are necessary to determine if these effects will also occur in other cell lines. Additional research is necessary to explore the role of total glutathione depletion in inducing tumor cell apoptosis.

In this study, isomerization conditions, cytotoxic activity, and stabilization of amygdalin from peach kernels were analyzed. Temperatures greater than 40 °C and pHs above 9.0 resulted in a quickly increasing isomer ratio (L-amygdalin/D-amygdalin). At acidic pHs, isomerization was significantly inhibited, even at high temperature. Ethanol inhibited isomerization; the isomer rate decreased with the ethanol concentration increasing. The growth-inhibitory effect on HepG2 cells of D-amygdalin was diminished as the isomer ratio increased, indicating that isomerization reduces the pharmacological activity of D-amygdalin. Extracting amygdalin from peach kernels by ultrasonic power at 432 W and 40 °C in 80% ethanol resulted in a 1.76% yield of amygdalin with a 0.04 isomer ratio. Hydrogel beads prepared by 2% sodium alginate successfully encapsulated the amygdalin, and its encapsulation efficiency and drug loading rate reached 85.93% and 19.21%, respectively. The thermal stability of amygdalin encapsulated in hydrogel beads was significantly improved and reached a slow-release effect in in vitro digestion. This study provides guidance for the processing and storage of amygdalin.

Concern about the side effects of liver cancer treatment has driven studies on anticancer to find compounds from plants that can act as chemotherapy. The anticancer activity ofChromolaena odorataagainst colorectal cancer, lung cancer, leukemia, cervical cancer, breast cancer, and liver cancer has been proven. However, this plant’s mechanism that can inhibit liver cancer cell growth is still undetermined. This study aims to investigate the anticancer activity ofC. odorataagainst HepG2 cells. Extraction ofC. odorataleaves was done by maceration method using 80% ethanol and further fractionated. Total flavonoid and major compound of the crude extract were determined by aluminum chloride colorimetric assay and Liquid Chromatography-Mass Spectrometry method. The IC50and proliferation analysis was performed by MTT assay. Cell cycle was analyzed by using flowcytometry. Total flavonoid of 1.95% and compounds such as 5,7,8,3ʹ,4ʹ-Pentamethoxyflavonone, 1-Carboethoxy-β-carboline, 3-Methylcanthin-2, 6- dion, Canthin-6-one were found inC. odorata. The proliferation of HepG2 was significantly lower after 72 hours of incubation with ½ IC50ofC. odoratafractions. HepG2 cells treated withC. odorataextract and fractions were accumulated in the G0-G1 phase. These results indicated thatC. odorataleaves could inhibit the proliferation of HepG2 cells and induce cell cycle arrest.

Background: Rhizoma Paridis (RP) is a traditional Chinese herb used for the treatment of tumors, detoxification and hemostasia. Studies show the main components of RP are Polyphyllin I (PPI), polyphyllin VI (PPVI), and polyphyllin VII (PPVII). However, the pharmaco-mechanisms of these compounds are not clear. Objective: By used 1H nuclear magnetic resonance (1H-NMR) based metabolomics approach to identify the Anticancer effects of PPI, PPVI and PPVII in HepG2 cells. Methods: 1H nuclear magnetic resonance (1H-NMR) based metabolomics approach was applied to investigate the toxicological effect of PPI, PPVI, PPVII on HepG2 cells. Multivariate statistical analysis was employed to examine the metabolic changes and abnormal metabolic pathways, including Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA), and orthogonal PLS-DA (OPLS-DA). Results: The results showed that the effects of metabolic phenotypes were affected separately by PPI, PPVI, and PPVII. The metabolic phenotypes were also changed over time. The characteristic metabolites were varied by affecting different polyphylins, which were identified by the reconstructed OPLSDA loading plots. According to the characteristic metabolites, the mainly disturbed metabolic pathways were found, such as alanine, aspartate and glutamate metabolism, pyruvate metabolism, glycine, serine, and threonine metabolism. Conclusion: The current work could allow us to understand the therapeutic effect of RP in metabolism. It also indicated that RP would be a promising candidate for liver cancer treatment.

Drug-drug cocrystals, which can regulate physicochemical properties of individual drugs and might produce synergistic therapeutic effects, have drawn growing interest in the pharmaceutical industry. In this study, a novel drug-drug (1:1) cocrystal hydrate of slightly water-soluble dihydromyricetin (DMY) and highly water-soluble pentoxifylline (PTX), DMY-PTX•H2O (1), was prepared by a slurry method. The single-crystal X-ray diffraction results reveal that the cocrystal is formed through hydrogen-bonding interactions between hydroxyl groups of DMY and four acceptors of PTX. The dynamic vapour sorption results indicate that the cocrystal displays reduced hydrophilicity compared with DMY. It is found that cocrystal formation narrows the solubility difference between two parent drugs. The equilibrium solubility of PTX decreases greatly, while that of DMY increases slightly. As a result, DMY and PTX are synchronously and sustainedly released from the cocrystal. Further, a synergistic anti-cancer effect of the cocrystal DMY-PTX•H2O (1) on HepG2 cells in vitro at a drug concentration of 100 μM was discovered. This study brings evidence of cocrystallization as a successful approach for synchronous sustained-release of two drugs with substantially different aqueous solubility.

Here in present work we studied the antibacterial and anticancer activity of gold nanoparticles (AuNPs), which are prepared through eco-friendly method using chloroauric acid and Pergulariadaemia leaves extract. The formation of the AuNPs and its structural features were fully examined by UV-Vis, SEM, HR-TEM and XRD analysis. The prepared AuNPs exhibited very high intense surface plasmon resonance (SPR) band in between 520 and 551 nm. SEM images showed AuNPs as spherical like morphology and HR-TEM images reveals the well-distributed AuNPs with average size of 30 nm. Further it was subjected to study antibacterial activity with various bacterial strains. The AuNPs coated cotton fabric has shown significant improvement in antibacterial activity against these strains. AuNPs coated cotton fabric display a superior UV-protection efficiency and it also exhibited finite anticancer response against HepG2 cell line.

Instead of Western blot being considered as a gold standard for intracellular protein expression assays, we developed a novel multiplexed high throughput (180 tests/day) in situ manual protein expression method directly in 96-well plates using 25,000–100,000 cells/well after formaldehyde fixation and Triton X 100 permeabilization. HepG2 cells were treated with ochratoxin A (OTA) and staurosporine (STP) to induce apoptosis. Antioxidant and apoptotic cell signaling protein expression were studied by various rabbit primary antibodies and HRP labeled secondary antibodies. The HRP labeled immune complexes were developed by H2O2/Ampliflu Red fluorogenic reagent and measured in a plate reader. Our assay can simultaneously quantify 22 protein antigens in one plate with 4 technical replicates with an interassay imprecision of

Metabolic disorders characterized by elevated blood glucose levels are a recognized risk factor for hepatocellular carcinoma (HCC). Lipid dysregulation is critically involved in the HCC progression, regulating energy storage, metabolism, and cell signaling. There is a clear link between de novo lipogenesis in the liver and activation of the NF-κB pathway, which is involved in cancer metastasis via regulation of metalloproteinases MMP-2/9. As conventional therapies for HCC reach their limits, new effective and safe drugs need to be found for the prevention and/or adjuvant therapy of HCC. The marine plant Posidonia oceanica (L.) Delile is endemic to the Mediterranean and has traditionally been used to treat diabetes and other health disorders. The phenol-rich leaf extract of Posidonia oceanica (POE) is known to have cell-safe bioactivities. Here, high glucose (HG) conditions were used to study lipid accumulation and fatty acid synthase (FASN) expression in human HepG2 hepatoma cells using Oil Red O and Western blot assays. Under HG conditions, the activation status of MAPKs/NF-κB axis and MMP-2/9 activity were determined by Western blot and gelatin zymography assays. The potential ameliorative role of POE against HG-related stress in HepG2 cells was then investigated. POE reduced lipid accumulation and FASN expression with an impact on de novo lipogenesis. Moreover, POE inhibited the MAPKs/NF-κB axis and, consequently, MMP-2/9 activity. Overall, these results suggest that P. oceanica may be a potential weapon in the HCC additional treatment.

The prevalence of nonalcoholic fatty liver disease (NAFLD) is estimated to be approximately about 25.24% of the population worldwide. NAFLD is a complex syndrome and is characterized by a simple benign hepatocyte steatosis to more severe steatohepatitis in the liver pathology. Phellinus linteus (PL) is traditionally used as a hepatoprotective supplement. Styrylpyrone-enriched extract (SPEE) obtained from the PL mycelia has been shown to have potential inhibition effects on high-fat- and high-fructose-diet-induced NAFLD. In the continuous study, we aimed to explore the inhibitory effects of SPEE on free fatty acid mixture O/P [oleic acid (OA): palmitic acid (PA); 2:1, molar ratio]-induced lipid accumulation in HepG2 cells. Results showed that SPEE presented the highest free radical scavenging ability on DPPH and ABTS, and reducing power on ferric ions, better than that of partitions obtained from n-hexane, n-butanol and distilled water. In free-fatty-acid-induced lipid accumulation in HepG2 cells, SPEE showed an inhibition effect on O/P-induced lipid accumulation of 27% at a dosage of 500 μg/mL. As compared to the O/P induction group, the antioxidant activities of superoxide dismutase, glutathione peroxidase and catalase were enhanced by 73%, 67% and 35%, respectively, in the SPEE group. In addition, the inflammatory factors (TNF-α, IL-6 and IL-1β) were significantly down-regulated by the SPEE treatment. The expressions of anti-adipogenic genes involved in hepatic lipid metabolism of 5’ adenosine monophosphate (AMP)-activated protein kinase (AMPK), sirtuin 1 (SIRT1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) were enhanced in the SPEE supplemented HepG2 cells. In the protein expression study, p-AMPK, SIRT1 and PGC1-α were significantly increased to 121, 72 and 62%, respectively, after the treatment of SPEE. Conclusively, the styrylpyrone-enriched extract SPEE can ameliorate lipid accumulation and decrease inflammation and oxidative stress through the activation of SIRT1/AMPK/PGC1-α pathways.

Dyslipidemia is a lipid metabolism disorder associated with the loss of the physiological homeostasis that ensures safe levels of lipids in the organism. This metabolic disorder can trigger pathological conditions such as atherosclerosis and cardiovascular diseases. In this regard, statins currently represent the main pharmacological therapy, but their contraindications and side effects limit their use. This is stimulating the search for new therapeutic strategies. In this work, we investigated in HepG2 cells the hypolipidemic potential of a picrocrocin-enriched fraction, analyzed by high-resolution 1H NMR and obtained from a saffron extract, the stigmas of Crocus sativus L., a precious spice that has already displayed interesting biological properties. Spectrophotometric assays, as well as expression level of the main enzymes involved in lipid metabolism, have highlighted the interesting hypolipidemic effects of this natural compound; they seem to be exerted through a non-statin-like mechanism. Overall, this work provides new insights into the metabolic effects of picrocrocin, thus confirming the biological potential of saffron and paving the way for in vivo studies that could validate this spice or its phytocomplexes as useful adjuvants in balancing blood lipid homeostasis.

The polysaccharides isolated and purified from different parts of the medicinal fungus Cordyceps cicadae were identified, and three extracts displaying significant biological activities were selected for further study. The bacterium substance polysaccharides (BSP), spore powder polysaccharides (SPP), and pure powder polysaccharides (PPP) were separated, purified, and collected from the sclerotia, spores, and fruiting bodies of Cordyceps cicadae, respectively. The structures of Cordyceps cicadae polysaccharides were analyzed using gas chromatography, Fourier-transform infrared spectroscopy, methylation analysis, and one-dimensional (1H and 13C) nuclear magnetic resonance spectroscopy. Moreover, the hypoglycemic effect of Cordyceps cicadae polysaccharides was examined in both in vitro and in vivo models. BSP, SPP, and PPP significantly increased glucose absorption in HepG2 cells, and alleviated insulin resistance (IR) in the in vitro model. SPP was the most effective, and was therefore selected for further study of its hypoglycemic effect in vivo. SPP effectively improved body weight and glucose and lipid metabolism in type 2 diabetes model mice, in addition to exerting a protective effect on liver injury. SPP regulated the mRNA expression of key PI3K/Akt genes involved in the insulin signaling pathway. The hypoglycemic mechanism of SPP may reduce hepatic insulin resistance by activating the PI3K/Akt signaling pathway. Spore powder polysaccharides (SPP) extracted from Cordyceps cicadae effectively improved body weight and glucose and lipid metabolism in type 2 diabetes model mice, in addition to exerting a protective effect on liver injury. The mechanism underlying the hypoglycemic effect of SPP regulates the mRNA expression of key PI3K/Akt genes involved in the insulin signaling pathway to alleviate insulin resistance. Our results provide a theoretical basis for research into the hypoglycemic effect of Cordyceps cicadae, and lay the foundation for the development of functional products.

Der Lebensstil der mit unserer schnelllebigen Welt einhergeht beeinflusst maßgebend die Gesundheit der Menschen. Nicht nur die Adipositas-Epidemie, sondern auch die Verbreitung der nicht-alkoholischen Fettleber-Erkrankung, NAFLD, stellen weltweite Herausforderungen für die moderne Medizin dar. Um ein besseres Verständnis über NAFLD zu bekommen werden die Effekte von NAFLD auf den Lipidstoffwechsel, in dem Apolipoproteine eine wesentliche physiologische Bedeutung einnehmen, untersucht. Ein kürzlich entdecktes Apolipoprotein, Apolipoprotein O, scheint im Lipidstoffwechsel eine Rolle zu spielen, und dadurch möglicherweise in der Pathobiochemie von NAFLD. Im Rahmen dieser Arbeit wurde der Effekt von Überernährung in Leberzellen simuliert. Dafür wurden HepG2 Zellen, eine humane Leberkrebszelllinie, herangezogen und mit Ölsäure inkubiert. Es erfolgte eine Lipidanreicherung, ähnlich wie bei NAFLD. Darüber hinaus wurde eine progressivere und fortgeschrittenere Form von NAFLD untersucht. Für dieses Stadium sind Entzündungen charakteristisch, die hier durch die Zugabe von Tumornekrosefaktor α und Menadion ausgelöst wurden. Schließlich wurde innerhalb dieser Bedingungen die Produktion von Apolipoprotein O untersucht. Eine Inkubation der Zellen mit 0.5 mM und 1 mM Ölsäure für 24 h Stunden führte zu einem etwa dreifach höheren Triglycerid-Spiegel verglichen mit der Kontrollprobe. Allerdings wurden deutlich erhöhte cytotoxische Effekte beobachtet, wenn die Zellen mit Ölsäure Konzentrationen über 0.5 mM versetzt wurden. Außerdem wurde festgestellt, dass ein Serumentzug von 24 h vor der Inkubation mit der Ölsäure die Triglycerid-Anreicherung möglicherweise fördert. Allgemein konnten nur geringe Effekte auf die Apolipoprotein O-Produktion gemessen werden. Trotzdem scheint eine Triglycerid-Anreicherung, wie sie nach 24 h Inkubation mit Ölsäure erreicht wurde, die Proteinproduktion zu fördern. Der Einsatz der zwei Stressfaktoren, Tumornekrosefaktor α und Menadion, beeinflusste den Triglycerid-Gehalt nicht, lieferte jedoch widersprüchliche Ergebnisse in Bezug auf die Apolipoprotein O-Synthese. Die ermittelten Daten deuten auf einen verringerten Gehalt an intrazellulären Apolipoprotein O durch die Anwesenheit der Stressfaktoren hin. Des Weiteren wurde die Aktivierung des nuklearfaktor kappa B Signalwegs durch Tumornekrosefaktor α und, zu einem kleineren Ausmaß, durch Menadion nachgewiesen. Schließlich wurden die Zellen mit einer Kombination aus Ölsäure und Stressfaktoren inkubiert, wodurch vermutlich eine geringe Inhibierung der Apolipoprotein O-Produktion stattfand. Darüber hinaus wurden mögliche protektive Effekte der Ölsäure beobachtet, die bei einer Simulation von Entzündungsreaktionen zu hinterfragen sind. Insgesamt bleibt es weiterhin schwierig eindeutige Aussagen über dem Zusammenhang von Überernährung und oxidativen Stress auf die Apolipoprotein O-Produktion zu treffen. In Zukunft gilt es das Gleichgewicht zwischen Lipidanreicherung und Lipidtoxizität zu finden, sowie geeignete Bedingungen zu definieren, die möglichst genau die zellulären Prozesse der Leber wiedergeben., Today’s fast-paced lifestyle has a major impact on human health and is the source of several disorders. Together with the obesity epidemic, the prevalence of non-alcoholic fatty liver disease, NAFLD, has become a worldwide challenge for modern medicine. Thus, it is crucial to gain a better understanding of NAFLD and its effect on lipid metabolism, in which apolipoproteins are of considerable physiological importance. A recently discovered apolipoprotein, apolipoprotein O, appears to be involved in lipid metabolism and consequently in the pathobiochemistry of NAFLD. In this thesis, the effect of overnutrition on liver cells was simulated by treating HepG2 cells, a human hepatoma cancer cell line, with oleic acid. Hence, lipid accumulation – as it occurs in NAFLD – is mediated. Furthermore, a more progressive form of NAFLD, which is characterized by inflammation, was mimicked by the addition of tumor necrosis factor α and menadione. Within this setting, the protein production of apolipoprotein O was studied. After exposing the cells for 24 h to 0.5 mM and 1 mM oleic acid, an estimated 3-fold increase of intracellular triglyceride content was observed. However, the cytotoxic impact on the cells drastically increased by the incubation with concentrations higher than 0.5 mM oleic acid. Furthermore, it was observed that serum starvation for 24 h before the treatment might support triglyceride accumulation. Even though only small effects of oleic acid on apolipoprotein O protein production were measured, it seems to be promoted by triglyceride deposition after 24 h of incubation with oleic acid. While tumor necrosis factor α and menadione did not affect the intracellular triglyceride level, contradictory outcomes were recorded regarding the effect on the protein content of apolipoprotein O. Overall, the data suggest a small decrease of apolipoprotein O protein production after the addition of the two stress factors. Moreover, the activation of the nuclear factor-kappa B pathway, monitored by two transcription factors, was achieved by tumor necrosis factor α and to a smaller extent also by menadione. When oleic acid and stress factors were simultaneously added to the cells, a slightly inhibited apolipoprotein O protein production might have been examined. However, the results do not reflect a uniform phenomenon. Nevertheless, possible protective properties of oleic acid in the inflammatory process have to be considered. Overall, assessing the impact of overnutrition and oxidative stress on the production of apolipoprotein O remains difficult. Further research on finding the balance between lipid accumulation and lipotoxicity as well as the reflection of the biochemical processes inside liver cells is required.

Background/aim: Ankaferd blood stopper (R) (ABS) is an herbal extract consisting of mixtures of Alpinia officinarum, Gycyrrhiza glabra, Vitis vinifera, Thymus vulgaris, and Urtica dioica plants and has been used in recent years in Turkish medicine as a hemostatic agent. Despite its extensive usage, there is no information available about the drug interaction in HepG2 cells. The current work evaluated the effect of ABS on the expression of CYP1A1-1A2, CYP2E1, and CYP3A4 isozymes that are primarily involved in drug and carcinogen metabolism. Materials and methods: We selected HepG2 cells as in vitro cellular models of the human liver. The cells were treated with different concentrations of ABS [0.25%-40% (v/v)]. A crystal violet staining assay was used to determine the cytotoxicity of ABS. We examined drug-metabolizing enzymes, including 7-ethoxyresorufin O-deethylase (CYP1A1), 7-methoxyresorufin O-demethylase (CYP1A2), aniline 4-hydroxylase (CYP2E1), and erythromycin N-demethylase (CYP3A4), in vitro in HepG2 cells. The expression (mRNA, protein) levels of drug-metabolizing enzymes were analyzed by qPCR and Western blotting, respectively. Results: The EC05 and EC10 values for ABS were 0.37% and 0.52% (v/v), respectively. Therefore, 0.37% and 0.52% (v/v) doses were used for the remaining portion of this study. Investigation of the expression and activity levels revealed that CYP1A1-1A2, CYP2E1, and CYP3A4 activities were not affected by ABS significantly, with qPCR and Western blot results corroborating this result. Conclusion: Our study found that the activity, mRNA, and protein expression levels of CYP isozymes did not change with the application of ABS, suggesting that when humans are exposed to ABS, there may not be any risk associated with clinical drug toxicity, cancer formation, and drug metabolism disorders in humans.

Hepcidin and ferritin are key proteins of iron homeostasis in mammals. In this study, we characterize a chimera by fusing camel hepcidin to a human ferritin H-chain to verify if it retained the properties of the two proteins. The construct (HepcH) is expressed in E. coli in an insoluble and iron-containing form. To characterize it, the product was incubated with ascorbic acid and TCEP to reduce and solubilize the iron, which was quantified with ferrozine. HepcH bound approximately five times more iron than the wild type human ferritin, due to the presence of the hepcidin moiety. To obtain a soluble and stable product, the chimera was denatured and renatured together with different amounts of L-ferritin of the H-chain in order to produce 24-shell heteropolymers with different subunit proportions. They were analyzed by denaturing and non-denaturing PAGE and by mass spectroscopy. At the 1:5 ratio of HepcH to H- or L-ferritin, a stable and soluble molecule was obtained. Its biological activity was verified by its ability to both bind specifically cell lines that express ferroportin and to promote ferroportin degradation. This chimeric molecule showed the ability to bind both mouse J774 macrophage cells, as well as human HepG2 cells, via the hepcidin–ferroportin axis. We conclude that the chimera retains the properties of both hepcidin and ferritin and might be exploited for drug delivery.

We determined the phenolic content and anti‐oxidation properties of ginger at different harvesting time and tested its effects on lipid droplet formation and glucose uptake in HepG2 cells. Ginger samples at different stages of maturity were harvested every two weeks starting from mid‐October for 16 weeks. Our data indicate that ginger has the highest phenolic contents and superior anti‐oxidation activity when harvested early (immature baby ginger); however, the concentration of phenolic contents and its anti‐oxidation activity were progressively reduced up to 50% as ginger matures. Furthermore, the data indicate that baby ginger extract inhibits lipid accumulation and triglyceride content in oleic acid‐induced HepG2 cells up to 20% in a dose‐dependent manner. Baby ginger exhibited significant inhibition of α‐amylase enzyme activity by 29.5% and ameliorated glucose uptake in HepG2 cell at similar level. Our results suggest that harvesting ginger at an appropriate (early) time may be beneficial for optimizing its biological active contents and qualitative properties. The data also suggest that a regular use of ginger can potentially lower incidences of obesity and diabetes., 6‐gingerol is the primary component in ginger; it is about 15‐fold and 53‐fold higher than 6‐paradol and 6‐shogaol, respectively. Consistent with TPC and antioxidant activity, the highest amount of these major phytochemicals was obtained when harvested at the first week; however, these phenolic compounds were gradually reduced to 50% of originals as ginger matured.

Aplydactylonins A-C (1–3), three new sesquiterpenes, were isolated from the Vietnamese sea hare Aplysia dactylomela. Their structures and absolute configurations were elucidated based on spectroscopic analysis, X-ray crystallography, and density functional theory (DFT) calculations of NMR and ECD data. Compound 2 exhibited cytotoxicity against HepG2, DU145 and A549 cells with respective IC50 values of 4.08 ± 0.63, 38.64 ± 1.04 and 12.33 ± 0.95 µM. In addition, HepG2 cells treated with 5 μM compound 2 for 48 h showed a significant increase in early apoptotic cells (P

Type 2 diabetes mellitus (T2DM), a chronic low-grade inflammatory disease with high morbidity and mortality, is a serious threat to public health. Previously we demonstrated that a traditional Chinese medicine formulation, Jiedu Tongluo Tiaogan Formula (JDTL), exerted a favorable hypoglycemic effect due to unknown molecular mechanisms involving interactions among JDTL compounds and various cellular components. This study aimed to explore JDTL mechanisms for alleviating hyperglycemia using an integrated strategy incorporating system pharmacology, bioinformatics analysis, and experimental verification. This strategy entailed initial elucidation of JDTL chemical composition using fingerprint analysis via high performance liquid chromatography (HPLC). Next, functions of putative shared target genes and associated pathways were deduced using GO and KEGG pathway enrichment and molecular docking analyses. Ultimately, targets associated with JTDL anti-T2DM effects were found to be functionally associated with biological functions related to lipopolysaccharide and cytokine receptor binding. These results implicated PI3K-Akt signaling pathway involvement in JDTL anti-T2DM effects, as this pathway had been previously shown to play significant roles in glucose and lipid metabolism-related diseases. Furthermore, addition of JDTL to INS-1 and HepG2 cell cultures stimulated cellular mRNA-level and protein-level expression leading to enhanced production of IRS1, Akt, and PI3K. In summary, here JDTL bioactive ingredients, potential targets, and molecular mechanisms underlying JDTL anti-T2DM effects were identified using a multi-component, multi-target, and multi-channel analytical approach, thus providing an important scientific foundation to facilitate development of new drugs mechanistic strategies for preventing and treating T2DM.

Spiroethers comprise a sub-class of spirocycles and exert various biological properties such as antibacterial and anti-tumor activities. The present study investigated the total spiroether isomers present in methanolic extract of root and aerial parts of 40 Arnebia species collected from 20 different geographical zones in Iran. Also, the cytotoxic effects of the methanolic extract of the samples and the retrieved isomers were assessed against eight cancerous and one non-cancerous cell-lines. In all Arnebia samples tested, two spiroether isomers namely, cis-(E)- and trans-(Z)-spiroethers were found and it was observed that root and aerial parts of Arnebia euchroma had significantly high levels of the two isomers as determined by High Performance Liquid Chromatography coupled with Photodiode Array Detection-Mass Spectrometry (HPLC-PAD-MS). Both isolated compounds, particularly cis-(E)-spiroether, could inhibit cancer cells proliferation while had little destructive effects in normal cells. Cis-(E)- and trans-(Z)-spiroethers inhibited HepG2 cell line proliferation with IC50 values of 3.23 ± 0.24 and 4.80 ± 0.15 µg/mL, respectively. Among the samples extracts, aerial parts and root extracts of A. euchroma had the highest anti-cancer activity against HepG2 with IC50 values of 6.32 ± 0.23 and 7.50 ± 0.11 µg/mL, respectively. The present study showed variations in cytotoxic activity of cis-(E)- and trans-(Z)-spiroethers among Arnebia species.

Introduction and Aim: Plants have become the current focus of research in treating the various diseases and ailments. Flacourtia jangomas (Lour.) Raeusch belongs to the familySalicaceae. Itis a small deciduous fruit tree having immense nutritional and medicinal significance. Different parts of the plant are pharmaceutically used forcuring various ailments. In this study, we investigated the hepatoprotective activity of Flacourtia jangomas (Lour.) Raeusch leaves and fruit methanolic extract on Paracetamol induced HepG2 cell line. Methods: The cytotoxic and hepatoprotective properties were evaluated by measuring cell viability; activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH); lipid peroxidation (malondialdehyde (MDA) levels). Results:The increased cell viability of 140.43± 4.07% and 133.93±3.20%was observed in HepG2 cells treated with methanolic extract of F. jangomas leaf and fruit extract respectively at 10µg/ml concentration and then decreased along with the rise of F. jangomas leaf and fruit extract concentrations. The level of LDH, ALT, AST and MDA decreased after F. jangomas leaf and fruit treatment compared to negative control. Conclusion: This study suggests that the methanolic Extract of F. jangomas (Lour.) Raeusch leaves(FJL)and fruit (FJF) shows hepatoprotective activity in Paracetamol induced HepG2 cell line by the decrease in AST and ALT activities and LDH and MDA level. Hence, it could be considered as a therapeutic agent in curing liver-related diseases.

The present study objective was to design and develop novel health-supplement formula from plant extracts and was to evaluate the formula for high episodic alcohol toxicities, and associated disorders against alcohol intoxicated and oxidative damaged Human Hepatoma HepG2 cell line.

This study was designed to evaluate the hepatoprotective effect of alcoholic and n-hexane extracts of crayfish Procambarus clarkii against CCl4 (4 mM) induced toxicity in the human hepatoma cell li...

To investigate the effect and mechanism of pinolenic acid (PNA) on H2O2‐induced oxidative stress injury in HepG2 cells. Methods: PNA was used to regulate oxidative stress injury of HepG2 cells induced by H2O2. Quantification of cell survival rate, accumulation of intracellular reactive oxygen species (ROS), and expression levels of anti‐oxidation‐related genes were determined using MTT, fluorescent probe technology (DCFH‐DA), and real‐time quantitative reverse transcription polymerase chain technology (qRT‐PCR) method, respectively. Meanwhile, the activity of intracellular antioxidant enzymes was determined by biochemical methods. The results showed that PNA improved the survival rate of HepG2 cells induced by H2O2 (29.59%, high‐dose group), reduced the accumulation of intracellular ROS (65.52%, high‐dose group), and reduced the level of intracellular malondialdehyde (MDA; 65.52%, high‐dose group). All these results were dose‐dependent, which indicated that PNA can improve oxidative stress damage of cells. Furthermore, the mechanism of PNA regulating oxidative stress was investigated from the gene level. Results showed that under supplementation of PNA, the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH‐Px) had been improved (39.74%, 17.58%, and 23.83%, high‐dose group). Further studies on gene expression which controls the activity of antioxidant enzymes showed that under the regulation of PNA, the expression level of Keap1 gene was decreased, while Nrf2 gene was increased. The expression levels of HO‐1 and NQO1 in the downstream of Nrf2 were increased. Results indicated that under the regulation of PNA, Nrf2 was separated from Keap1, entered the nucleus, bound to ARE, and up‐regulated the expression levels of HO‐1 and NQO1 genes. Conclusion: PNA has a conspicuous improvement effect on oxidative stress damage induced by H2O2 in HepG2 cells. We also found the antioxidant mechanisms of PNA where it protected cells from oxidative stress damage by causing nuclear translocation of Nrf2 gene and up‐regulated the expression levels of antioxidant enzymes in the downstream. This shows that PNA prevented oxidative stress by mediating the Keap1/Nrf2 transcriptional pathway and down‐regulating enzyme activities.

Xiaoyu Wang,1 Guangbing Li,1,2 Changfa Guo,3 Jiayao Zhang,1 Junjie Kong,1,2 Jingyi He,1,2 Feiyu Li,1 Yong Liu,1 Yang Yang,1 Ziwen Lu,1 Jun Liu1,2 1Department of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, People’s Republic of China; 2Department of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People’s Republic of China; 3Department of Neurosurgery, Shandong Provincial Hospital Affiliated to Shandong University, Cheeloo College of Medicine, Jinan, Shandong, People’s Republic of ChinaCorrespondence: Jun LiuDepartment of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, People’s Republic of ChinaEmail dr_liujun1967@126.comBackground: The AMP-activated protein kinase alpha (AMPKα) pathway has widely been considered a key factor in energy metabolism. Ethyl 2-[2,3,4-trimethoxy-6-(1-octanoyl)phenyl] acetate (TMPA) is a novel AMPK agonist, which influences the stability of Nuclear Receptor Subfamily 4, Group A, Member 1 (Nur77)-serine-threonine kinase 11 (LKB1) in the nucleus. A recent study has determined that TMPA can ameliorate the reduction of insulin resistance in type II db/db mice. However, the role of TMPA in hepatocyte lipid metabolism has not been elucidated.Objective: To investigate whether TMPA could ameliorate liver lipid accumulation under the stimulation of free fatty acids (FFAs) in vitro.Methods: We evaluated differences of Nur77 and AMPK pathway in mice fed a high-fat diet and those fed a normal diet. In vitro, TMPA was added to HepG2 cells and primary hepatocytes before FFAs stimulation. Oil red O staining, Nile red staining were used to evaluate lipid deposition. Western blot and immunofluorescence were used to quantify related proteins.Results: Nur77, AMPKα, LKB1, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), acetyl-CoA carboxylase phosphorylation (p-ACC), and carnitine palmitoyltransferase 1 (CPT1A) showed significant differences in vivo. Under the intervention of TMPA, HepG2 cells and primary hepatocytes showed considerable amelioration of lipid deposition and improved the expression of phosphorylated (p)-AMPKα (p-AMPKα), p-LKB1, p-ACC, and CPT1A. Furthermore, Western blotting and immunofluorescence studies indicated that LKB1 dramatically increased expression in the cytoplasm but decreased in the nucleus. Further, AMPKα phosphorylation (p-AMPKα) also showed a higher expression in cytoplasm instead of the nucleus.Conclusion: TMPA ameliorated lipid accumulation by influencing the stability of Nur77-LKB1 in vitro.Keywords: ethyl 2-[2,3,4-trimethoxy-6-(1-octanoyl)phenyl] acetate, Nur77, AMPK pathway, HepG2 cells, primary hepatocytes

Xiaoyu Wang,1 Guangbing Li,1,2 Changfa Guo,3 Jiayao Zhang,1 Junjie Kong,1,2 Jingyi He,1,2 Feiyu Li,1 Yong Liu,1 Yang Yang,1 Ziwen Lu,1 Jun Liu1,2 1Department of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, People’s Republic of China; 2Department of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People’s Republic of China; 3Department of Neurosurgery, Shandong Provincial Hospital Affiliated to Shandong University, Cheeloo College of Medicine, Jinan, Shandong, People’s Republic of ChinaCorrespondence: Jun LiuDepartment of Hepatobiliary Surgery and Center of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, People’s Republic of ChinaEmail dr_liujun1967@126.comBackground: The AMP-activated protein kinase alpha (AMPKα) pathway has widely been considered a key factor in energy metabolism. Ethyl 2-[2,3,4-trimethoxy-6-(1-octanoyl)phenyl] acetate (TMPA) is a novel AMPK agonist, which influences the stability of Nuclear Receptor Subfamily 4, Group A, Member 1 (Nur77)-serine-threonine kinase 11 (LKB1) in the nucleus. A recent study has determined that TMPA can ameliorate the reduction of insulin resistance in type II db/db mice. However, the role of TMPA in hepatocyte lipid metabolism has not been elucidated.Objective: To investigate whether TMPA could ameliorate liver lipid accumulation under the stimulation of free fatty acids (FFAs) in vitro.Methods: We evaluated differences of Nur77 and AMPK pathway in mice fed a high-fat diet and those fed a normal diet. In vitro, TMPA was added to HepG2 cells and primary hepatocytes before FFAs stimulation. Oil red O staining, Nile red staining were used to evaluate lipid deposition. Western blot and immunofluorescence were used to quantify related proteins.Results: Nur77, AMPKα, LKB1, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), acetyl-CoA carboxylase phosphorylation (p-ACC), and carnitine palmitoyltransferase 1 (CPT1A) showed significant differences in vivo. Under the intervention of TMPA, HepG2 cells and primary hepatocytes showed considerable amelioration of lipid deposition and improved the expression of phosphorylated (p)-AMPKα (p-AMPKα), p-LKB1, p-ACC, and CPT1A. Furthermore, Western blotting and immunofluorescence studies indicated that LKB1 dramatically increased expression in the cytoplasm but decreased in the nucleus. Further, AMPKα phosphorylation (p-AMPKα) also showed a higher expression in cytoplasm instead of the nucleus.Conclusion: TMPA ameliorated lipid accumulation by influencing the stability of Nur77-LKB1 in vitro.Keywords: ethyl 2-[2,3,4-trimethoxy-6-(1-octanoyl)phenyl] acetate, Nur77, AMPK pathway, HepG2 cells, primary hepatocytes

Epidemiological studies have postulated an inverse correlation between developing cancer and neurodegeneration. It is known that the secretome plays a vital role in cell–cell communication in health and disease; the microglia is the resident macrophage of the central nervous system which maintains neuronal integrity by adapting as the microenvironment changes. The present study aimed to identify, in a cell model, biomarkers that link neurodegenerative diseases to cancer or vice versa. Real-time PCR and western blot analysis were used to characterize the effects on gene and protein expression of human hepatoblastoma (HepG2) and human microglia (HMC-III) cells after exchanging part of their conditioned medium. Biomarkers of the endoplasmic reticulum, and mitophagy and inflammatory processes were evaluated. In both cell types, we observed the activation of cytoprotective mechanisms against any potential pro-oxidant or pro-inflammatory signals present in secretomes. In contrast, HepG2 but not HMC-III cells seem to trigger autophagic processes following treatment with conditioned medium of microglia, thus suggesting a cell-specific adaptive response.

Combinations of anti-cancer drugs can overcome resistance to therapy and provide new more effective treatments. In this work we have analyzed the effect of the polyphenol quercetin and the anti-cancer sphingosine analog fingolimod on the sphingolipid metabolism in HepG2 cells, since sphingolipids are recognized as mediators of cell proliferation and apoptosis in cancer cells. Treatment of hepatocellular carcinoma HepG2 cells with quercetin and fingolimod, alone or in combination, induced different degrees of sphingomyelin (SM) reduction and a corresponding activation of neutral sphingomyelinase (nSMase). Western blot analysis showed that only treatments containing quercetin induced up-regulation of nSMase expression. The same treatment caused elevation of ceramide (CER) levels, whereas the observed alterations in sphingosine (SPH) content were not statistically significant. The two tested drugs induced a reduction of the pro-proliferative sphingolipid, sphingosine 1 phosphate (S1P), in the following order: quercetin, fingolimod, quercetin + fingolimod. The activity of the enzyme responsible for CER hydrolysis, alkaline ceramidase (ALCER) was down-regulated only in the incubations involving quercetin and fingolimod did not affect this activity. The enzyme, maintaining the balance between apoptosis and proliferation, sphingosine kinase 1 (SK1), was down-regulated by incubations in the following order: quercetin, fingolimod, quercetin + fingolimod. Western blot analysis showed down-regulation in SK1 expression upon quercetin but not upon fingolimod treatment. Studies on the effect of quercetin and fingolimod on the two proteins associated with apoptotic events, AKT and Bcl-2, showed that only quercetin, alone or in combination, down-regulated the activity of the two proteins. The reported observations provide information which can be useful in the search of novel anti-tumor approaches, aiming at optimization of the therapeutic effect and maximal preservation of healthy tissues.

Diabetes mellitus leads to cellular damage and causes apoptosis by oxidative stress. Heartwood extract of Pterocarpus marsupium has been used in Ayurveda to treat various diseases such as leprosy, diabetes, asthma, and bronchitis. In this study, we worked out the mechanism of the antidiabetic potential of methanolic heartwood extract of Pterocarpus marsupium (MPME). First, metabolic profiling of MPME was done using gas chromatography-mass spectrometry (GCMS), ultra-performance liquid chromatography-mass spectroscopy (UPLC-MS), and high-performance thin-layer chromatography (HPTLC) to identify phenols, flavonoids, and terpenoids in MPME. Biological studies were carried out in vitro using the HepG2 cell line. Many antidiabetic compounds were identified including Quercetin. Methanolic extract of MPME (23.43 µg/mL–93.75 µg/mL) was found to be safe and effective in reducing oxyradicals in HepG2 cells. A concentration of 93.75 µg/mL improved glucose uptake efficiently. A significant decrease in oxidative stress, cell damage, and apoptosis was found in MPME-treated HepG2 cells. The study suggests that the heartwood of Pterocarpus marsupium offers good defense in HepG2 cells against oxidative stress and improves glucose uptake. The results show the significant antidiabetic potential of MPME using a HepG2 cell model. The effect seems to occur by reducing oxidative stress and sensitizing the cells towards glucose uptake, hence lowering systemic glucose levels, as well as rescuing ROS generation.

Autophagy plays a critical role in many vitally important physiological and pathological processes, such as the removal of damaged and aged organelles and redundant proteins. Although autophagy is mainly a protective process for cells, it can also cause cell death. In this study, we employed in situ and ex situ surface-enhanced Raman scattering (SERS) spectroscopies to obtain chemical information of lysosomes of HepG2 cells. Results reveal that the SERS profiles of the isolated lysosomes are different from the in situ spectra, indicating that lysosomes lie in different microenvironments in these two cases. We further investigated the molecular changes of isolated lysosomes according to the autophagy induced by starvation viaex situ SERS. During autophagy, the conformation of proteins and the structures of lipids have been affected, and autophagy-related molecular evidence is given for the first time in the living lysosomes. We expect that this study will provide a reference for understanding the cell autophagy mechanism.

A new fluorescent probe (PAC) based on phenothiazine was designed and synthesized for the detection of hydrazine (N2H4), which uses methyl cyanoacetate as the recognition group and phenothiazine as the fluorescent chromophore. The Stoke shift of probe PAC at the presence of hydrazine reached to 120 nm and the detection limit of probe PAC performed as 6.7 ppb. PAC had AIE properties and provided with the functions of naked-eye recognition. The DFT and TD-DFT calculations exhibited that the probes of PAC performed good responses to hydrazine. In addition, cell imaging showed that the probe PAC effectively recognized the hydrazine in HepG2 cells and stained the cells green.

Chemical investigation of the whole plant of Sauropus hirsutus Beille led to the isolation of eight quinolines and two known flavonoids. Furthermore, five quinolines were new, two were reported in plant for the first time and one was known. Cytotoxicity evaluation against cholangiocarcinoma, KKU-M156, showed that the most active compound was 4-hydroxy-6-methoxy-7,8-methylenedioxyquinaldine (IC50 20.54 ± 6.82 µM) which was a little more active than the cisplatin standard (IC50 24.39 ± 1.14 µM).

Glucose-regulated protein 78 (GRP78) is overexpressed in a wide variety of solid tumors, serving as a well-characterized target for tumor imaging or therapy. In this work, we developed a GRP78-responsive radiotracer (DOTA-68Ga)-Gly-Gly-Gly-Ser-Phe-Ser-Ile-Ile-His-Thr-Pro-Ile-Leu-Pro-Leu-Gly-Gly-Cys ([68Ga]SP94) for hepatocellular carcinoma (HCC) micro-PET imaging. DOTA-SP94 was synthesized by solid phase synthesis and then radiolabeled with 68GaCl3 with >99% radiochemical purity. The expression levels of GRP78 in HepG2 cells were confirmed by Western blotting. In vitro and in vivo study of [68Ga]SP94 showed high stability and high uptake in GRP78-overexpressing HepG2 cells and tumor, fast clearance, and low nontarget uptake. Micro-PET images showed excellent tumor accumulation of [68Ga]SP94 in the HepG2-implanted nude mice tumor model. Additionally, the radiotracer uptake in HepG2 tumors can be blocked by unlabeled DOTA-SP94, suggesting that the tracer uptake by HCC was receptor-mediated. We envision that our radiotracer can be used for noninvasive imaging of HCC and is worthy of further clinical investigations.

High-mobility group box 1 (HMGB1), a highly conserved chromosome protein, is considered as a potential therapeutic target and novel biomarker because of its regulation in the proliferation and metastasis of Hepatocellular carcinoma (HCC). Calenduloside E (CE), a natural active product, has been reported to anti-cancer effect. However, the role and underlying molecular mechanism of CE in HCC is still unclear. The purpose of this study is to investigate the effects of CE on the proliferation and migration of HCC, and then explore the possible underlying molecular mechanism. HepG2 cells were treated with CE or transfected with HMGB1 shRNA plasmids, EdU and colony formation assays were used to detect cell proliferation ability. Wound healing and transwell assays were used to determine the role of CE in cell migration. The expression of Cyclins, PCNA, MMPs, HMGB1, N-cadherin, E-cadherin and phosphorylation of p38, ERK and JNK were all detected using Western blotting. Our results showed that CE inhibited HepG2 cells proliferation and migration in a dose dependent manner; reduced the expression levels of Cycins, PCNA, HMGB1, MMPs and N-cadherin; up-regulated E-cadherin expression; enhanced the phosphorylation of p38 and JNK signalling pathways. Blocking the activation of p38 and JNK obviously reversed CE-mediated inhibitory effects on HepG2 cell proliferation and migration; reversed CE-induced down-regulation of Cyclins, PCNA, MMPs, N-cadherin and HMGB1, as well as E-cadherin up-regulation. In conclusion, our study suggested that CE reduces the expression levels of Cyclins, MMPs and epithelial-mesenchymal transformation (EMT) through p38/JNK-HMGB1 signaling axis and then inhibits HepG2 cells proliferation and migration in HepG2 cells. This study provides a new perspective for the anti-tumour molecular mechanism of CE in HCC.

Epigenetic regulation by histone acetyltransferase (HAT) is associated with various biological processes and the progression of diseases, including nonalcoholic fatty liver disease (NAFLD). The objective of this study was to investigate whether the hypolipidemic properties of black mulberry (

A previously undescribed anthraquinone-aurone adduct 1, named 8-(4′-hydroxy-3′,5′-dimethoxyphenyl)-12-hydroxy-7-(6′′,7′′-dimethylfuran-2′′-one)-9,16-anthraquinone-furochromene-3,2′′-dione, was isolated from the fruits of Cucumis bisexualis. Its structure was determined by comprehensive spectroscopic data and references. Compound 1 was evaluated for its inhibitory effect against the increase in AST and ALT levels in H2O2-induced HepG2 cells in the range of their maximum nontoxic concentration. Compound 1 exhibited moderate hepatoprotective activities in vitro.

Phytochemical evaluation of the alcoholic extract of Rubia cordifolia Linn. roots resulted in the identification of two anthraquinone compounds 1a (1,4-dihydroxyanthracene-9,10(4aH,9aH)-dione) and 1b (2-methylanthracene-9,10(4aH,9aH)-dione). The isolated compounds were characterized by spectroscopic methods and their structures were established. The derivatives were found to be nontoxic against Vero and HepG2 cells.

Flavonoids from common buckwheat hulls (BHFs) show significant antioxidant and antidiabetic potential. However, their hepatoprotective property is yet to be defined. This study aims to examine the hepatoprotective effect of BHFs in type 2 diabetes mellitus (T2DM) rats and chronic high glucose‐damaged HepG2 cells. Results showed that BHF treatment significantly relieves the state of insulin resistance, thereby reducing blood glucose and improving oxidative stress in T2DM rats. It is worth mentioning that BHF treatment improved diabetes‐induced liver damage disorders, manifested as the clearance of liver fat and the decline of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. In vitro, HepG2 cells pretreated with BHFs maintained higher superoxide dismutase (SOD), glutathione peroxidase (GSH‐px), and catalase (CAT) activities than the unprotected group. In parallel, compared with the unprotected group, BHFs significantly reduced the leakage of ALT and AST in pre‐protected group dose‐dependently. These results indicated that BHFs had considerable antioxidant and hepatoprotective potential and could be promising to be used as nutraceuticals and dietary supplements to prevent and/or protect against liver disorders., Buckwheat hull flavonoids had a considerable ability to regulate blood glucose. Buckwheat hull flavonoids showed strong antioxidant activity in vivo and in vitro. Buckwheat hull flavonoids played a hepatoprotective role in vivo and in vitro.

The study aimed to investigate the bioaccessibility and the stability of purified ethanol extracts (PEE) from Cudrania cochinchinensis roots, and the changes in antioxidant activity and enzyme inhibition activity and cytotoxicity by in vitro digestion. No compounds were lost during the gastrointestinal digestion (GID) process, 15 compounds were tentatively identified using UPLC-TOF-MS/MS technology, including five phenolic and ten flavonoid compounds. After the gastric digestion (GD), the bioaccessibility of phenolic and flavonoid compounds was 55.66% and 52.44%, respectively. After the intestinal digestion (ID), the bioaccessibility of phenolic and flavonoid compounds reduced to 39.70% and 33.36%, respectively. The gastrointestinal digestion (GID) decreased the bioaccessibility of phenolic compounds. All compounds were affected throughout the gastrointestinal digestion (GID), among which the more affected compounds were 4-hydroxybenzoic acid, eriodictyol-7-O-glucoside, resveratrol and 3,6,3’,4’-tetrahydroxyflavone (compared with the peak intensity of the undigested PEE). Furthermore, there was a reduction in the antioxidant activity, inhibition activity against α-glucosidase and tyrosinase, and cytotoxicity toward HepG2 cells of the PEE after GID. There was a significantly positive correlation between the bioactivity and the total phenolic/flavonoid contents. The results showed that in vitro digestion had significant effects on the phenolic content and bioactivity. Although phenolic contents were decreased during GID, C. cochinchinensis roots is still a good source of bioactive compounds for the development of functional foods with health benefits.

Two new triterpenoids, entanolide (1) and methyl 3,4-secotirucalla-23-oxo-4(28),7,24-trien-21-al-3-oate (2), together with nine known compounds (3-11), were isolated from the bark of Entandrophragma angolense. Their structures were elucidated based on spectroscopic analyses, mainly 1 D and 2 D NMR spectral data. Compounds 1-6 and 8 were evaluated for their cytotoxicity against HepG2 cells, and compounds 2-5 exhibited weak activities.

New effective, economical and safe antimalarial drugs are urgently needed due to the development of multi-drug-resistant strains of the parasite. Homeopathy uses ultra-diluted doses of various substances to stimulate autoregulatory and self-healing processes to cure various ailments. The aim of the study was to evaluate the in vitro and in vivo antimalarial efficacy of a homeopathic drug, Chininum sulphuricum 30C.In vitro antiplasmodial activity was screened against the P. falciparum chloroquine-sensitive (3D7) strain, and cell viability was assessed against normal human dermal fibroblasts and HepG2 cells. Suppressive, preventive and curative studies were carried out against P. berghei-infected mice in vivo.Chininum sulphuricum (30C) revealed good antiplasmodial activity in vitro, with 92.79 ± 6.93% inhibition against the 3D7 strain. The cell viability was 83.6 ± 0.6% against normal human dermal fibroblasts and 95.22 ± 5.1% against HepG2 cells. It also exhibited suppressive efficacy with 95.56% chemosuppression on day 7 with no mortality throughout the follow-up period of 28 days. It also showed preventive activity against the disease. Drug treatment was also safe to the liver and kidney function of the host as evidenced by biochemical studies.Chininum sulphuricum 30C exhibited considerable antimalarial activity along with safety to the liver and kidney function of the host.Hintergrund: Es besteht dringender Bedarf an neuen wirkungsvollen, ökonomischen und sicheren Malariamedikamenten angesichts der Entwicklung multiresistenter Stämme des Parasiten. In der Homöopathie werden hochverdünnte Dosen verschiedener Substanzen eingesetzt, um selbstregulierende und selbstheilende Prozesse zur Heilung verschiedener Leiden anzustoßen. Das Ziel dieser Studie war es, die Wirksamkeit des homöopathischen Präparats Chininum sulfuricum 30C gegen Malaria in vitro und in vivo zu beurteilen. Methoden: Die antiplasmodiale Aktivität in vitro wurde an P. falciparum, chloroquinsensitiver Stamm (3D7) getestet, und die Zellvitalität wurde bei normalen humanen Dermis-Fibroblasten sowie HepG2-Zellen untersucht. In vivo wurden suppressive, präventive und kurative Studien bei P.-berghei-infizierten Mäusen durchgeführt. Ergebnisse: Chininum sulfuricum (30C) zeigte in vitro gute antiplasmodiale Aktivitätmit 92,79 ± 6,93% Inhibition des 3D7-Stamms. Die Zellvitalität lag bei 83,6 ± 0,6% bei normalen humanen dermalen Fibroblasten bzw. 95,22 ± 5,1% bei HepG2-Zellen.Außerdem zeigte das Präparat suppressive Wirksamkeit mit 95,56% Chemosuppression an Tag 7 ohne Mortalität im gesamten Nachbeobachtungszeitraum von 28 Tagen. Auch zeigte es präventive Wirksamkeit gegen die Krankheit. Die Behandlung war dabei sicher in Bezug auf die Leber- und Nierenfunktion des Wirts, wie biochemische Studien belegen. Schlussfolgerung: Chininum sulfuricum 30C zeigte erhebliche Aktivität gegen die Malariainfektion und war dabei sicher in Bezug auf die Leber- und Nierenfunktion des Wirts.

Cisplatin (CDDP) has been successfully used in chemotherapy for liver cancer. However, the development of CDDP resistance in HepG2 cells usually leads to relapse and a worsening prognosis. MiR-340-5p has attracted much attention because of its ability to affect cell resistance. This project is intended to explore the role of miR-340-5p and CDK6 in CDDP-R HepG2 cells and provide new ideas for the treatment of liver cancer. A dual-luciferase reporter assay was used to confirm the targeting relationship between miR-340-5p and CDK6. We constructed a CDDP-resistant model of HepG2 cells to examine the effect of miR-340-5p on the drug sensitivity of HepG2 cells. CDDP-R HepG2 cells were transfected with miR-340-5p overexpression plasmid and CDK6 silencing plasmid. QRT-PCR was used to detect the expression of miR-340-5p and CDK6. A western blot was performed to determine the expression of CDK6, CyclinD1, and CyclinD2. CCK-8, flow cytometry, TUNEL and Clonogenic assays were also carried out to detect CDDP-R HepG2 cells. There is a targeting relationship between miR-340-5p and CDK6. The drug resistance of CDDP-R HepG2 cells was significantly higher than that of CDDP-S HepG2 cells. CDDP-R HepG2 cells transfected with both miR-340-5p overexpressing plasmid and CDK6 silencing plasmid showed a lower proliferation ability, cell cycle arrest in the G0/G1 phase, and lower drug resistance compared with single CDDP-R HepG2 cells. Overexpression of miR-340-5p aggravated CDDP-R HepG2 cells' apoptosis and inhibited cell viability. Overexpression of miR-340-5p could reverse the sensitivity of HepG2 cells to CDDP by inhibiting the expression of CDK6 in HepG2 cells.

Ruthenium complexes are at the forefront of developments in metal-based anticancer drugs, but many questions remain open regarding their reactivity in biological media, including the role of transferrin (Tf) in their transport and cellular uptake. A well-known anticancer drug, KP1019 ((IndH)[RuIIICl4(Ind)2], where Ind = indazole) and a reference complex, [RuIII(nta)2]3− (nta = nitrilotriacetato(3−)) interacted differently with human apoTf, monoFeTf, or Fe2Tf. These reactions were studied by biolayer interferometry (BLI) measurements of Ru–Fe–Tf binding to recombinant human transferrin receptor 1 (TfR1) in conjunction with UV-vis spectroscopy and particle size analysis. Cellular Ru uptake in human hepatoma (HepG2) cells was measured under the conditions of the BLI assays. The mode of Tf binding and cellular Ru uptake were critically dependent on the nature of Ru complex, availability of Fe(III) binding sites of Tf, and the presence of proteins that competed for metal binding, particularly serum albumin. Cellular uptake of KP1019 was not Tf-mediated and occurred mostly by passive diffusion, which may also be suitable for treatments of inoperable cancers by intratumoral injections. High cellular Ru uptake from a combination of [RuIII(nta)2]3− and Fe2Tf in the absence of significant Ru–Tf binding was likely to be due to trapping of Ru(III) species into the endosome during TfR1-mediated endocytosis of Fe2Tf.

Duguetia pycnastera Sandwith (Annonaceae) is a tropical tree that can be found in the Guyanas, Bolivia, Venezuela, and Brazil. In Brazil, it is popularly known as “ata”, “envira”, “envira-preta”, and “envira-surucucu”. In the present work, we investigated the in vitro and in vivo HepG2 cell growth inhibition capacity of D. pycnastera leaf essential oil (EO). The chemical composition of the EO was determined by GC–MS and GC–FID analyses. The alamar blue assay was used to examine the in vitro cytotoxicity of EO in cancer cell lines and non-cancerous cells. In EO-treated HepG2 cells, DNA fragmentation was measured by flow cytometry. The in vivo antitumor activity of the EO was assessed in C.B-17 SCID mice xenografted with HepG2 cells treated with the EO at a dosage of 40 mg/kg. Chemical composition analysis displayed the sesquiterpenes α-gurjunene (26.83%), bicyclogermacrene (24.90%), germacrene D (15.35%), and spathulenol (12.97%) as the main EO constituents. The EO exhibited cytotoxicity, with IC50 values ranging from 3.28 to 39.39 μg/mL in the cancer cell lines SCC4 and CAL27, respectively. The cytotoxic activity of the EO in non-cancerous cells revealed IC50 values of 16.57, 21.28, and >50 μg/mL for MRC-5, PBMC, and BJ cells, respectively. An increase of the fragmented DNA content was observed in EO-treated HepG2 cells. In vivo, EO displayed tumor mass inhibition activity by 47.76%. These findings imply that D. pycnastera leaf EO may have anti-liver cancer properties.