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• Magnoflorine, a natural isoquinoline alkaloid was experimentally analyzed by different spectroscopies techniques. • The optimized structural and electronic properties of the molecule were predicted by DFT calculations. • Vibrational frequencies assignments were performed based on PED calculations. • The reactivity sites were evaluated by theoretical calculation. • The preference of anxiolytic activity toward the extracellular in comparison with intracellular domains of GABAA receptor was predicted. Magnoflorine (MGF), an isoquinoline alkaloid, emerges as a quaternary aporphine alkaloid derived from L-tyrosine amino acid, found in families Magnoliaceae plants and it presents important biological activity being noted for its effect on the nervous system. In this work, a deep study of structural, vibrational and electronic properties has been carried out for the MGF. From Potential Energy Surface (PES) scan the most stable conformer of alkaloid was identified and geometrical parameters were determined by Density Functional Theory (DFT) using B3LYP/6–311++G** method. A complete vibrational assignment of the normal modes was theoretical and experimentally achieved from FT-IR and Raman spectroscopies and scaled SQM methodology. Electronic transitions observed in UV–visible spectrum in aqueous medium were identified using TD-DFT methodology, with the π→π* transition being the most probable UV signal. The MEPs, the electric charge distribution along with the HOMO and LUMO frontier orbitals were analyzed and the GAP energy values justified the stability of the molecule in aqueous medium. The molecular electrostatic potential map reveals the most favourable region for electrophilic attack on MGF. Molecular docking was used to analyze the anxiolytic biological activity of the title molecule on the GABA A receptor. The results suggest an intermolecular binding capability of MFG toward both intracellular and extracellular domains of GABA A receptor, and the extracellular domain presents a higher affinity by alkaloid. Finally, the biological study infers that MGF could be used as a potential anxiolytic compound. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] This study systematically explores a green extraction process of T. crispa stems to selectively isolate therapeutic compounds, borapetoside C (BPC) and magnoflorine (MGF), recognized for their anti-diabetic properties. In agreement with Hansen solubility parameters prediction, H 2 O and EtOH were found to be suitable solvents for extraction of BPC and MGF, respectively, resulting in 33 % and 45 % extractabilities after 60 min. Further investigations demonstrated considerable increase in BPC extractability using 20 % EtOH:H 2 O mixture at 40 °C, with complete extraction achieved after 60 min. While for MGF, pure-EtOH at 40 °C, was the most suitable solvent, showing the highest selectivity and complete extraction after 100 min. The BPC-rich extract from 20 % EtOH:H 2 O exhibited higher anti-diabatic activity (IC 50 = 0.63 ± 0.03 and 0.72 ± 0.04 mg/mL, respectively, for α-glucosidase and α-amylase enzymes inhibition activity), and considerably lower cytotoxicity to L6 and HepG2 cells, with IC 50 = 0.26 ± 0.16 and 0.24 ± 0.02 mg/mL, respectively, compared with the MGF-rich extract obtained with pure-EtOH. Based on these results, a sequential extraction scheme was proposed involving initial pure-EtOH extraction to selectively and completely remove MGF, followed by extraction with a 20 % EtOH:H 2 O mixture to recover the remaining BPC, which was approximately 80 % of the BPC originally present. The obtained MGF-free BPC-rich extract showed significantly lower cytotoxicity (IC 50 = 0.31 ± 0.06 mg/mL against L6 cell) and higher enzyme inhibition activities (IC 50 = 0.53 ± 0.32 and 0.52 ± 0.02 mg/mL for α-glucosidase and α-glucosidase enzymes inhibition activity), comparable to acarbose (IC 50 = 0.43 ± 0.02 and 0.83 ± 0.03 mg/mL for α-glucosidase and α-glucosidase enzymes inhibition activity), the result that potentially leads to the development of a promising industrial process to harness T. crispa for diabetes prevention and treatment. [ABSTRACT FROM AUTHOR]

Abstract Objective To investigate efficacy of Chinese medicine magnoflorine combined with hyaluronic acid (HA)-gel in promoting subchondral bone (SCB) regeneration and attenuating cartilage degeneration in early osteoarthritis (OA). Methods MC3T3-E1 under magnoflorine treatment was assayed by XTT to determine cell viability. Cell proliferation was reflected through cell cycle. Osteoblast mineralization was stained by Alizarin Red. Standardized bone canal of 1 mm in diameter and 4 mm in depth was made on tibial medial plateau of 4-month-old Dunkin-Hartley spontaneous knee OA guinea pigs. Guinea pigs (n = 5/group) were treated once intra-bone canal injection of 2 μl HA-gel, 2 μl HA-gel+50 ng magnoflorine and null (Defect) respectively, except sham group. The left hind limbs were harvested for μCT scan and histopathological staining 2-month post-surgery. Results 25 μg/ml magnoflorine treatment significantly increased cell viability, S-phase and mineralization of MC3T3-E1 cells. In vivo, HA-gel + magnoflorine treatment significantly altered SCB microstructure; changes included increase in bone volume fraction (BV/TV), trabecular number (Tb.N), connectivity density (Conn.Dn), and decrease in degree of anisotropy (DA), which implied trabecular bone regeneration. Treatment also resulted in a decrease in modified Mankin's scores, and an increase in volume ratio of hyaline cartilage (HC)/calcified cartilage (CC) and fractal dimension (FD, roughness indicator of osteochondral conjunction), compared to Defect and HA groups. Furthermore, FD was positively associated with volume ratio of HC/CC and negatively associated with modified Mankin's scores. Finally, histological results showed that due to a faster regeneration of SCB with the HA-gel + magnoflorine treatment, the reduction of cartilage matrix and the decreased expression of chondrogenic signals were attenuated. Conclusion Our study elucidated the potential benefits of HA-gel + magnoflorine in promoting SCB regeneration and revealed a protective effect of stimulating recovery of the SCB integrity on attenuating cartilage degradation to prevent OA progression. Highlights • Recovery subchondral bone integrity is pivotal to attenuate cartilage degeneration. • Magnoflorine might directly stimulate osteoblast differentiate to calcified osteoid. • Magnoflorine prevents OA progression by preventing subchondral bone loss. [ABSTRACT FROM AUTHOR]

The in vivo immunomodulatory activities of Tinospora crispa have been reported but its molecular mechanisms underlying its immunomodulatory properties remains obscure and the active constituents contributing to the activities have not been identified. The present study was aimed to investigate the immunomodulatory effects of T. crispa extract (TCE) and its chemical constituents on RAW 264.7 macrophages. Six known compounds including magnoflorine and syringin were isolated by various chromatographic techniques from TCE and their structures were determined spectroscopically. A validated HPLC method was used to quantify magnoflorine and syringin in the extract. The immunomodulatory effects of TCE and its isolated compounds on chemotaxis, phagocytosis, production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines which include tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and monocyte chemoattractant protein-1 (MCP-1) on macrophages were assessed. TCE increased the chemotaxis and phagocytic activity of macrophages and significantly enhanced the production of ROS, NO and pro-inflammatory cytokines. All alkaloids isolated, specifically magnoflorine showed remarkable inducing effects on the chemotaxis, phagocytic activity, ROS and NO productions and the secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. In contrast, syringin potently reduced the chemotaxis, phagocytic activity, ROS and NO productions and secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. TCE showed strong immunostimulant effects on various components of the immune system and these activities were possibly contributed mainly by the alkaloids specifically magnoflorine. TCE has potential to be developed as an effective natural immunostimulant for improvement of immune-related disorders. [ABSTRACT FROM AUTHOR]

Cognitive deficit is the main clinical feature of Alzheimer's disease (AD), and the massive death of neuronal cells is the leading cause of cognitive deficits. So, there is an urgent clinical need to discover effective drugs to protect brain neurons from damage in order to treat AD. Naturally-derived compounds have always been an important source of new drug discovery because of their diverse pharmacological activities, reliable efficacy and low toxicity. Magnoflorine is a quaternary aporphine alkaloid, which naturally exist in some commonly used herbal medicines, and has good anti-inflammatory and antioxidant effects. However, magnoflorine has not been reported in AD. To investigate the therapeutic effect and mechanism of magnoflorine on AD. Neuronal damage was detected by flow cytometry, immunofluorescence and western blotting. Oxidative stress was measured by detection of SOD and MDA, as well as JC-1 and reactive oxygen species (ROS) staining. The APP/PS1 mice were given drugs by intraperitoneal injection (I.P.) every day for one month, and then the new object recognition and Morris water maze were used to detect the cognitive ability of the mice. We demonstrated that magnoflorine reduced Aβ-induced PC12 cell apoptosis and intracellular ROS generation. Further studies found that magnoflorine significantly improved cognitive deficits and AD-type pathology. Most interestingly, the efficacy of magnoflorine was better than that of the clinical control drug donepezil. Mechanistically, based on RNA-sequencing analysis, we found that magnoflorine significantly inhibited phosphorylated c-Jun N-terminal kinase (JNK) in AD models. This result was further validated using a JNK inhibitor. Our results indicate that magnoflorine improves cognitive deficits and pathology of AD through inhibiting of JNK signaling pathway. Thus, magnoflorine may be a potential therapeutic candidate for AD. [Display omitted] [ABSTRACT FROM AUTHOR]

An LC–MS/MS method was developed and validated for the simultaneous determination of magnoflorine, berberrubine, jatrorrhizine, coptisine, epiberberine, palmatine and berberine in human urine. The sample preparation procedure involved the four-fold dilution of the urine samples with acetonitrile/water (1:3, v/v). The chromatographic separation was achieved on a Hedera ODS-2 column under gradient elution at a flow rate of 0.4 mL/min with acetonitrile and water containing 0.5% formic acid as the mobile phase. The mass detection was performed in the positive mode. Calibration curves of the seven alkaloids showed good linearity (correlation coefficients > 0.9973) over their concentration ranges. To meet the requirements of urinary excretion study for each alkaloid in human, the lower limit of quantification was set at different values from 0.05063 ng/mL to 2.034 ng/mL for the seven alkaloids, respectively. The intra- and inter-batch precision and accuracy were all within ±15%. No matrix effect was observed for the analytes. The validated method was applied to the excretion study for the seven alkaloids in healthy Chinese volunteers after oral administration of Shuanghua Baihe tablets. The average 72 h cumulative urinary excretion of magnoflorine, berberrubine, jatrorrhizine, coptisine, epiberberine, palmatine and berberine accounted for 1.81%, 0.27%, 0.29%, 0.046%, 0.027%, 0.010% and 0.021% of the respective administered dose. [ABSTRACT FROM AUTHOR]

[Display omitted] • Magnoflorine stimulated Th1/Th2 cytokines, T and B lymphocytes proliferation. • It stimulated phagocytosis, NO production and serum levels of lysozyme and MPO. • CD4+ and CD8+ cell T population was significantly enhanced by magnoflorine. • It upregulated swelling rate of mice paw in delayed type hypersensitivity reaction. • It upregulated anti-sRBC immunoglobulins expression levels in treated mice. Magnoflorine shows a diverse range of pharmacological actions, including immunomodulatory, antioxidant and neuropharmacological activities. However, its effects on the immune responses in animal studies have not been reported. In this study, magnoflorine isolated from Tinospora crispa, at doses of 25, 50 and 100 mg/kg was administered to male Balb/c mice daily for 14 days to evaluate its effect on innate immune responses, while for evaluation of adaptive immune responses, on day 0 the mice were injected intraperitoneally with sheep red blood cells (sRBC) and treated orally with the various doses of magnoflorine for the same duration. The effects of magnoflorine on phagocytosis, myeloperoxidase (MPO) activity, lysozyme serum level, nitric oxide (NO) production, CD4+ and CD8+ cells population, T and B lymphocytes proliferation, activated T cells cytokines production, antibodies levels and delayed type hypersensitivity (DTH) were determined. Magnoflorine dose-dependently stimulated NO production, E. coli engulfment by neutrophils and peritoneal macrophages, MPO activity and lysozyme serum level in treated mice. Magnoflorine at 100 mg/kg exhibited comparable stimulation of B cell production compared to levamisole at 2.5 mg/kg. It also significantly increased CD4+ and CD8+ cells population, upregulated the Th1 (IFN-γ, IL-2 and TNF-α) and Th2 (IL-4 and IL-6) cytokines in a dose-dependent manner. At similar concentrations, magnoflorine also exhibited a strong dose-dependent stimulation on DTH reaction and upregulation of immunoglobulins (IgG and IgM) production in mice immunized with sRBC. The strong upregulation of innate and adaptive immune responses indicates that magnoflorine has potential to be developed into an effective immunostimulant. [ABSTRACT FROM AUTHOR]

Background: As a prolonged autoimmune disorder, rheumatoid arthritis (RA) is characterised by synovial hyperplasia and the erosion of bone and cartilage. Magnoflorine (MAG) is the main component purified from Clematis manshurica Rupr. Recent studies have shown that MAG has anti-inflammatory, antioxidant, and immunosuppressive effects, which are relevant to anti-RA activities.Objective: The current investigation was conducted to explore the anti-RA effects of MAG and to discover the possible molecular mechanisms.Methods: In vitro experiments, CCK-8, wound healing, and transwell assays were utilized to evaluate the anti-proliferative, anti-migratory, and anti-invasive activities of MAG, respectively. The rate of cell distribution and cell apoptosis were evaluated by flow cytometry. ROS generation was detected by DCFH-DA staining. Western blotting, quantitative real-time polymerase chain reaction assay, and immunofluorescent staining were employed to test the anti-RA effect of MAG as well as to explore the potential mechanisms by evaluating related gene and protein expression. For in vivo experiments, an adjuvant-induced arthritis (AIA) rat model was established. The related parameters were measured in rats. Then, rats were sacrificed, and ankle joints were collected for histopathological analysis and observation.Results: MAG significantly decreased the proliferation, migration, invasion, and reactive oxygen species levels in IL-1β-treated MH7A cells. Furthermore, MAG promoted cell apoptosis by increasing Bax levels and decreasing Bcl-2 levels. MAG also induced cell cycle arrest. Inflammatory cytokines (iNOS, COX-2, IL-6, and IL-8) and MMPs (MMP-1, 2, 3, 9, and 13) were reduced by MAG treatment. Molecular analysis revealed that MAG exerted anti-RA effects by partly inhibiting the PI3K/Akt/NF-κB signalling axis and activating the Keap1-Nrf2/HO-1 signalling pathway. In vivo studies have revealed that MAG treatment substantially improved severe symptoms in AIA rats, and these curative effects were linked to the attenuation of inflammatory responses.Conclusion: These results first suggested that MAG exhibits anti-arthritic effects in IL-1β-treated MH7A cells and AIA rat models. Thus, MAG may be used as a new drug to treat RA clinically. [ABSTRACT FROM AUTHOR]

This article presents the isolation, quantification and antioxidant evaluation of bioactive principles from Epimedium elatum . LC–MS guided isolation technique was applied for the separation of target constituents. Three isolates; magnoflorine, chrysin and dibenzylideneacetone (DBA) were isolated for the first time from E. elatum using LC–MS guided isolation method. Nine natural products, viz. icariin, epimedoside A, epimedin A, epimedin B, epimedin C, ikarisoside C, baohuoside II, magnoflorine and chrysin were simultaneously quantified by reverse phase HPLC-UV-DAD method. The HPLC method was validated in terms of precision and accuracy. Excellent specificity and linearity within test ranges for all standard calibration curves having regression coefficient of different linear equations in the range of 0.9966–0.9999 were observed. Relative recovery rates varied between 98.09 ± 0.44 and 105.34 ± 1.89% with relative standard deviation of less than 3%. This modified HPLC method is in accordance with yinyanghuo. All the 10 isolated constituents were screened for DPPH radical scavenging activity. Dibenzylideneacetone (DBA) turned out to be the most potent isolate with IC 50 of 4.32 μM. [ABSTRACT FROM AUTHOR]

Abstract: Antidiabetic potential of Tinospora cordifolia stem is well proven. In the course of screening of useful α-glucosidase inhibitors, we prepared alkaloid fraction (AFTC) and isolated three isoquinoline alkaloids, namely, jatrorrhizine, palmatine and magnoflorine as active candidates for α-glucosidase inhibition. The enzyme kinetics was studied using sucrose and maltose as substrates. Michaelis–Menten constant (K m) and maximal velocity (V max) values were estimated. A significant decrease in V max and unaltered K m was observed in case of jatrorrhizine and palmatine (non-competitive inhibition). Magnoflorine was found to increase apparent K m and shown to be reversible, competitive inhibition. The IC50 value as sucrase inhibitor was 36.25, 23.46 and 9.8μg/mL for jatrorrhizine, palmatine and magnoflorine, respectively, and as maltase inhibitor was 22.05, 38.42 and 7.6μg/mL for jatrorrhizine, palmatine and magnoflorine, respectively. In vivo studies were conducted on rats to determine oral glucose tolerance test (OGTT), using different substrates: glucose, sucrose and maltose. The increase in plasma glucose level was significantly suppressed (P <0.01) by all the three alkaloids at 20mg/kg b.w. Magnoflorine possessed the most potential activity as α-glucosidase inhibitor in vitro and in vivo. [Copyright &y& Elsevier]

[Display omitted] • MF has excellent fluorescent properties and can accurately reflect the metabolic distribution of this drug in cells. • MF can be observed accumulating in the mitochondrial area. • The mitochondrial target of MF, ClO-, was revealed in the nano-sized visualization area. • MF shows a definite response and binds to its target, ClO-, in ferroptosis. In current practice, drug visualization strategies mainly include 3H-or 14C-modification, or dye-labeling steps. Compared to current drug labeling strategies, drug molecules with autofluorescence can achieve accurate visualization of the drug's subcellular distribution. To this end, we screened various compounds in the traditional Chinese medicine compound library and selected a natural, label-free, fluorescent drug molecule named magnoflorine (MF). MF has fluorescent properties and does not require external intervention by the current labeling strategies, thereby proving to be suitable for reporting its distribution in living cells using structured illumination microscopy (SIM). In addition, using SIM, we found that MF not only had a high quantum yield but could also be well localized to the mitochondria. More importantly, the binding target of MF in mitochondria, namely hypochlorite (ClO-), was also revealed for the first time at the nanoscale visualization level. Finally, we also found that MF can play a role in binding to the ClO- as a target during ferroptosis, hence indicating that MF is a possible intervention drug for this process. In conclusion, we have identified for the first time a new fluorescent molecule, MF, that allows visualizing accurate drug distribution in organelles without additional labeling strategies with SIM. Furthermore, we have discovered the binding target of MF, which is beneficial for understanding of regulatory mechanism of drugs in various diseases. [ABSTRACT FROM AUTHOR]

Roots and barks of various Berberis species are used as folk remedy for the treatment of various inflammatory diseases such as lumbago, rheumatism and to reduce fever. Six isoquinoline alkaloids namely berberine, berbamine, palmatine, oxyacanthine, magnoflorine, and columbamine were isolated as the main components of alkaloidal fraction from the roots of Turkish Berberis species and effects were studied using various in vivo models in mice. All alkaloids inhibited inflammations in varying degrees, among them berberine, berbamine and palmatine were shown to possess significant and dose-dependent inhibitory activity against serotonin-induced hind paw oedema both on oral and topical applications and acetic acid-induced increase in vascular permeability on oral administration. Moreover, these three alkaloids were also shown to possess dose-dependent antinociceptive activity, which assessed by using the model based on the inhibition of p-benzoquinone-induced writhing movements as well as antipyretic activity on FCA-induced increased rectal temperature on subacute administration. However, all alkaloids induced gastric lesions in varying degrees. [Copyright &y& Elsevier]

Berberis dictyophylla F., a famous Tibetan medicine, has been used to prevent and treat diabetic retinopathy (DR) for thousands of years in clinic. However, its underlying mechanisms remain unclear. The present study was designed to probe the synergistic protection and involved mechanisms of berberine, magnoflorine and berbamine from Berberis dictyophylla F. on the spontaneous retinal damage of db/db mice. The 14-week spontaneous model of DR in db/db mice were randomly divided into eight groups: model group, calcium dobesilate (CaDob, 0.23 g/kg) group and groups 1–6 (different proportional three active ingredients from Berberis dictyophylla F.). All mice were intragastrically administrated for a continuous 12 weeks. Body weight and fasting blood glucose (FBG) were recorded and measured. Hematoxylin-eosin and periodic acid–Schiff (PAS) stainings were employed to evaluate the pathological changes and abnormal angiogenesis of the retina. ELISA was performed to assess the levels of IL-6, HIF-1α and VEGF in the serum. Immunofluorescent staining was applied to detect the protein levels of CD31, VEGF, p-p38, p-JNK, p-ERK and NF-κB in retina. In addition, mRNA expression levels of VEGF, Bax and Bcl-2 in the retina were monitored by qRT-PCR analysis. Treatment with different proportional three active ingredients exerted no significant effect on the weight, but decreased the FBG, increased the number of retinal ganglionic cells and restored internal limiting membrane. The results of PAS staining demonstrated that the drug treatment decreased the ratio of endothelial cells to pericytes while thinned the basal membrane of retinal vessels. Moreover, these different proportional active ingredients can markedly downregulate the protein levels of retinal CD31 and VEGF, and serum HIF-1α and VEGF. The gene expression of retinal VEGF was also suppressed. The levels of retinal p-p38, p-JNK and p-ERK proteins were decreased by drug treatment. Finally, drug treatment reversed the proinflammatory factors of retinal NF-κB and serum IL-6, and proapoptotic Bax gene expression, while increased antiapoptotic Bcl-2 gene expression. These results indicated that DR in db/db mice can be ameliorated by treatment with different proportional three active ingredients from Berberis dictyophylla F. The potential vascular protection mechanisms may be involved in inhibiting the phosphorylation of the MAPK signaling pathway, thus decreasing inflammatory and apoptotic events. [Display omitted] [ABSTRACT FROM AUTHOR]

Tinospora cordifolia (TC) is being used as a blood purifier in Ayurveda since ancient time. It is a very popular immunomodulator and holds anti-inflammatory and anti-oxidative potential, hence anti-aging properties. Therefore, it is also known as 'Amrita' in Ayurveda and is widely used to treat diabetes mellitus type II (T2DM) and its secondary complications; however, its underlying mechanism was not expedited to date. To explore the in vivo therapeutic efficiency and mechanism of action of TC and its secondary constitute magnoflorine on the skeletal muscle atrophy in the rat model of T2DM. Animal model of T2DM was developed using streptozotocin (STZ) injection followed by intervention with TC, metformin, and magnoflorine for three weeks. Confirmation of T2DM and abrogation of atrophic markers and possible mechanisms on supplementation of TC and magnoflorine were explored using histology, bio-assays, Western blotting, and q-PCR. TC and Magnoflorine supplementations significantly (p ≤ 0.05) decreased the fasting blood glucose (FBG) levels in T2DM rats. Both treatments prevented the lean body, individual skeletal muscle mass, and myotubes diameter loss (p ≤ 0.05). Magnoflorine significantly reduced the degradation of the protein indicated by biochemical markers of atrophy i.e. decreased serum creatine kinase (CK) levels and increased myosin heavy chain-β (MyHC-β) levels in muscles. Q-PCR and western blotting supported the findings that magnoflorine significantly increased the mRNA and protein abundances (~3 fold) of MyHC-β.TC and magnoflorine efficiently decreased the expression of ubiquitin-proteasomal E3-ligases (Fn-14/TWEAK, MuRF1, and Atrogin 1), autophagy (Bcl-2/LC3B), and caspase related genes along with calpains activities in T2DM rats. Both TC and magnoflorine also increased the activity of superoxide dismutase, GSH-Px, decreased the activities of β-glucuronidase, LPO, and prevented any alteration in the catalase activity. In contrast, magnoflorine increased expression of TNF-α and IL-6 whereas TC and metformin efficiently decreased the levels of these pro-inflammatory cytokines (p ≤ 0.05). However, magnoflorine was found to increase phosphorylation of Akt more efficiently than TC and metformin. TC, and magnoflorine are found to be effective to control fasting blood glucose levels significantly in T2DM rats. It also promoted the Akt phosphorylation, suppressed autophagy and proteolysis that might be related to blood glucose-lowering efficacy of magnoflorine and TC. However, increased muscle weight, specifically of the soleus muscle, expression of IL-6, and slow MyHC indicated the increased myogenesis in response to magnoflorine and independent from its hypoglycemic activity. Image 1 [ABSTRACT FROM AUTHOR]

Magnoflorine is an essential type of alkaloid and possesses anti-tumor activity in multiple cancers. Recent studies have demonstrated that magnoflorine plays tumor-suppressive roles in gastric and breast cancers. However, its role in osteosarcoma (OS) tumorigenesis is enigmatic. This study aimed to investigate the role and mechanism of magnoflorine in OS. Two human OS cells (MG-63 and U-2 OS) were treated with different concentrations of magnoflorine. Cell viability and invasion were then detected by Cell Counting Kit-8 and Transwell assay, respectively. And the effects of magnoflorine on the epithelial-mesenchymal transition (EMT) and cisplatin sensitivity were also measured. To explore the potential mechanism, we assayed the influence of magnoflorine on the miR-410-3p/HMGB1/NF-κB signaling pathway. Additionally, rescue experiments were performed to further confirm the regulation mechanism of magnoflorine. Magnoflorine inhibited the viability, invasion, and EMT of OS cells in a dose-dependent manner. And it increased the sensitivity of OS cells to cisplatin. Magnoflorine significantly suppressed HMGB1 expression and NF-κB activation, but upregulated miR-410-3p level. Overexpression of HMGB1 promoted NF-κB activation and reversed the effects of magnoflorine on the viability, invasion, EMT and cisplatin sensitivity of OS cells. miR-410-3p mimic inhibited the EMT of OS cells, which was restored by HMGB1 upregulation. And miR-410-3p inhibitor abrogated the influence of magnoflorine on HMGB1 expression in OS cells. Magnoflorine inhibited the malignant phenotypes and increased cisplatin sensitivity of OS cells via modulating miR-410-3p/HMGB1/NF-κB pathway. These results indicated that magnoflorine might be a novel drug for the treatment of OS. [ABSTRACT FROM AUTHOR]

According to the Chinese medicine, magnoflorine exerted significant anti-inflammatory effects and potentially promoted synthesis of proteoglycans in chondrocytes to reverse the progression of rheumatoid arthritis. However, the latent beneficial effect of magnoflorine for the treatment of traumatic osteoarthritis (OA) is still unknown. Therefore, we aim to demonstrate the efficacy of magnoflorine combined with HA-gel in attenuating cartilage degeneration in anterior cruciate ligament transection (ACLT) induced OA rat model. We found that the histological results showed the elevated cartilage matrix, chondrogenic signals and chondroprogenitor cells in HA-gel + magnoflorine treatment. HA-gel + magnoflorine treatment resulted in a decreased modified Mankin's score, and a higher volume ratio of hyaline cartilage (HC)/calcified cartilage (CC) and HC/Sum (whole cartilage), compared to ACLT and HA-gel groups. Furthermore, both the volume ratios of HC/Sum and HC/CC were negatively correlated with modified Mankin's scores. Finally, HA-gel + magnoflorine could significantly increase the BV/TV, Tb.Th, and decrease the Tb.Pf, Po(tot), Conn.Dn and Tb.Sp. In vitro , 50 μg/ml magnoflorine treatment could significantly increase the viability, S-phase, migration rate and chondrogenesis of chondroprogenitor cells. There were significant downregulations of MAPK/NF-κB signaling, and upregulations of chondrogenic signals in 50 μg/ml magnoflorine treatment. There were significant downregulations of proinflammatory cytokines and upregulation of IL-10 in HA-gel + magnoflorine treated group. Therefore, our study elucidated a protective effect of HA-gel + magnoflorine on attenuating cartilage degradation and maintaining SCB stabilization in ACLT induced OA. [ABSTRACT FROM AUTHOR]

• A method to quantitate Berberis components and berberrubine is described. • Adjusting collision energy gave good linearity for all analytes. • One complete analysis required only 20 μL of mouse serum and 15 min. Berberidis cortex , the dry bark of Berberis L., is used to treat diabetes and contains at least three bioactive components: berberine (BBR), berbamine (BBM) and magnoflorine (MGF). BBR in turn is metabolized into berberrubine (BRB). Although it is possible to quantify each of these components individually in serum, there are currently no methods for simultaneously quantifying all four. Here, we developed a specific and rapid method for simultaneously quantifying BBR, BBM, MGF and BRB in mouse serum using ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were pretreated by protein precipitation, separated using an ACQUITY UPLC® BEH C18 column and detected by a triple quadrupole mass spectrometer with electrospray ionization. The compound [9,10-(OC2H 3) 2 ]-BBR (d 6 -BBR) was used as internal standard for BBR and BRB, boldine (BOL) for MGF and tetrandrine (TET) for BBM. The m / z transitions for precursor/product ion pairs were 336.1/320.2 for BBR, 305.2/566.3 for BBM, 342.0/297.1 for MGF, 322.1/307.2 for BRB, 342.2/294.3 for d 6 -BBR, 312.2/580.3 for TET and 328.1/265.2 for BOL. We validated our method in terms of selectivity, linearity and lower limit of quantification, accuracy, precision, matrix effect and recovery, dilution integrity and stability. This method showed good linearity from 0.1 to 40 ng/mL for BBR, 8 to 3200 ng/mL for BBM, 5 to 2000 ng/mL for MGF and 0.2 to 80 ng/mL for BRB. The chromatographic run time was 3.9 min, and sample preparation took approximately 15 min per batch. Finally, we used our method to measure BBR, BBM, MGF and BRB in serum from diabetic mice after gavage administration of BBR hydrochloride, BBM hydrochloride, and MGF. This method is precise, accurate and suitable for high-throughput sample analysis. [ABSTRACT FROM AUTHOR]

• A validated UHPLC-UV-MS method developed for distinguishing T. crispa from related species. • Intra- and inter-day RSD for replicates were 0.9 – 6.8 % and 1.2 – 9.1 %, respectively. • Limits of detection ranged from 0.3 to 10 μg/mL. • The data suggests two dietary supplements claim to contain T. cordifolia as mislabeled. Tinospora species are morphologically similar. Several cases of human toxicity have been reported in association with T. crispa. A chemical fingerprint was developed to differentiate T. crispa from its closely related species and to quantitate its major furanoditerpenes namely as borapetosides B, C and F. The rapid, sensitive and repeatable method was established using ultra–high performance liquid chromatography coupled with photodiode array and single quadrupole electrospray mass spectrometry detectors using a flavonoid, two alkaloids, an amide and six diterpenoids. Qualitative and quantitative determination was performed by UHPLC-UV and confirmed by MS. The intra-day RSD for replicates was between 0.9 and 6.8% and inter-day RSD was between 1.2 and 9.1%. Recovery was 97–103 %. The method is useful to achieve decisiveness in not only identifying but also differentiating T. crispa from T. sinensis and other closely related Tinospora species. Seventeen Tinospora plant samples and seventeen dietary supplements claiming T. crispa , T. sinensis and T. cordifolia were analyzed. The newly developed and validated method successfully resulted in the conclusive identification of two dietary supplements to be mislabeled. [ABSTRACT FROM AUTHOR]

• Magnoflorine from Rhizoma coptidis attenuated DNCB-induced atopic dermatitis-like symptoms in mice. • Magnoflorine inhibited IFN-γ and TNF-α-induced apoptosis of HaCaT cell. • Magnoflorine attenuated the expressions of apoptosis-related proteins in HaCaT cells. • Magnoflorine increased mitochondrial membrane potential. In Sheng Nong's herbal classic in China, Rhizoma coptidis a RC: Rhizoma coptidis a (RC) could be used to treat Atopic dermatits b AD: Atopic dermatits b (AD), but its core ingredient(s) and mechanism remains unknown. The present study aimed to find out the ingredients against AD and expound its mechanisms. Seven alkaloids were isolated from RC to compare the inhibition against HaCaT cells by MTT assays and apoptosis of cells stimulated with TNF-α/IFN-γ by flow cytometry. The effects of target alkaloids against AD were evaluated on DNCB c DNCB: 2,4-dinitrochlorobenzene c (2,4-dinitrochlorobenzene)-induced atopic dermatitis in mice. Seven alkaloids were isolated from RC successfully. The results from MTT and flow cytometry indicated that among these alkaloids, only magnoflorine d MAG: magnoflorine d (MAG) had no obvious toxicity on cells, but could inhibit the apoptosis of the cells stimulated with TNF-α/IFN-γ. Further animal experiments confirmed that MAG significantly attenuated the AD-like symptom and inhibited the AD-induced increases in IgE/IL-4, as compared with control (P < 0.01). Moreover, MAG reduced the low Δψ m e Δψ m : mitochondrial membrane potential e (mitochondrial membrane potential) in HaCaT cells. The results of western blotting proved that MAG inhibited apoptosis of keratinocytes through decreasing the expressions of CTSB f CTSB: cathepsin B f (cathepsin B), Cyte C g Cyte C: cytochrome C g (cytochrome C), Bid and caspase-3/7/8/9. Overall, MAG inhibited apoptosis by decreasing the expression of apoptotic pathway-related proteins, and laid a foundation for the study of AD mechanisms. [ABSTRACT FROM AUTHOR]

Breast cancer is a leading cause of cancer death among women worldwide. Doxorubicin (DOX) is a broad-spectrum anti-breast cancer agent, but its clinical use is restricted due to irreversible tissue toxicity. Thereby, new therapeutic approaches are urgently required to promote the sensitivity of breast cancer cells to DOX. Magnoflorine (Mag), a quaternary alkaloid isolated from Chinese herb Magnolia or Aristolochia, has various biological activities, such as anti-inflammation, anti-cancer, and anti-anxiety. In the study, we explored the effects Mag on the sensitivity of breast cancer cells to DOX. We demonstrated that Mag strongly promoted DOX-induced anti-proliferative effects in breast cancer cells while not in normal cells. Mag addition markedly promoted the effects of DOX on the inhibition of migration and invasion in breast cancer cells. DOX-triggered DNA damage in breast cancer cells was further accelerated by combination with Mag. DOX-induced cell distribution in G2/M phase was markedly elevated when co-treated with Mag. Additionally, DOX/Mag combinational treatment significantly induced apoptosis in breast cancer cells when compared to DOX alone group through inducing Caspase-3 cleavage. Moreover, Mag markedly promoted the role of DOX in autophagy induction by elevating light chain 3 (LC3)-II expression. Combination treatment with DOX and Mag significantly inhibited the activation of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling, and promoted p38 mitogen-activated protein kinase (MAPK) pathway. In addition, treatment with wortmannin (Wor, a blocker of autophagosome formation) markedly reduced DOX/Mag-induced p38 MAPK activation and LC3 conversion in breast cancer cells. Further, in MCF-7 xenograft model, DOX combined with Mag displayed a significant anti-tumor effect with little toxicity to organs such as liver, heart, kidney and spleen. These findings suggested that Mag promoted the anti-cancer effects of DOX to induce cellular apoptosis and autophagy in breast cancer cells. [ABSTRACT FROM AUTHOR]