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The purpose of this study was to determine the role of HYP-PDT and its cytotoxic and immunomodulatory efficacy in the treatment of oral cancer. SCC-25(oral squamous cell carcinoma) and HGF-1 (gingiva fibroblasts) cells were properly prepared and then treated with HYP-PDT. After HYP-PDT treatment, changes in the concentrations of sTNF-R1, sTNF-R2 and sCD30 were analyzed using Bioplex method. For SCC-25 cells: after HYP-PDT, there was a marked increase in the secretion of sTNF-R1 and sCD30 and no change in concentration of sTNF-R2. The applied HYP-PDT showed no effect on the secretion of sTNF-R1, sTNF-R2 and sCD30 by the HGF-1 cell line. HYP-PDT shows immunomodulatory potential in in vitro experiments. [ABSTRACT FROM AUTHOR]

The aim of the study is to evaluate the cytotoxic and immunomodulatory effects of HYP-PDT in vitro studies on cultures of selected skin cell lines, as a possibility of using PDT for patients with atopic dermatitis and psoriasis. Primary Dermal Fibroblast and Primary Epidermal Keratinocytes was used in the study. After incubation the cells were treated with hypericin, exposed to VIS visible light. Then cell viability was determined by the MTT assay and cytokines markings were done with Bio-Plex Pro™ Assay kit and with Bio-Plex Suspension Array System. The experiment showed a significant cytotoxic effect of HYP-PDT and in sublethal doses, an immunomodulatory effect manifested in changes in the concentrations of selected cytokines The obtaining results confirming the destructive effect of HYP- PDT on the selected, skin cells lines show a possibility of extending the indication for HYP-PDT, for patients with inflammatory skin diseases. [ABSTRACT FROM AUTHOR]

[Display omitted] Rheumatoid arthritis (RA) is a joint-destructive autoimmune disease that severely affects joint function. Despite the variability of treatment protocols, all of them are associated with severe side effects that compromise patient compliance. The main aim of the current study is to prepare localized effective RA treatment with reduced side effects by combining nanoencapsulation, photodynamic therapy (PDT) and hollow microneedles (Ho-MNs) to maximize the pharmacological effects of hypericin (HYP). To attain this, HYP-loaded emulsomes (EMLs) were prepared, characterized and administered through intradermal injection using AdminPen™ Ho-MNs combined with PDT in rats with an adjuvant-induced RA model. The prepared EMLs had a spherical shape and particle size was about 93.46 nm with an absolute entrapment efficiency. Moreover, confocal imaging indicated the interesting capability of Ho-MNs to deposit the HYP EMLs to a depth reaching 1560 µm into the subcutaneous tissue. In vivo, study results demonstrated that the group treated with HYP EMLs through Ho-MNs combined with PDT had no significant differences in joint diameter, TNF-α, IL1, HO-1, NRF2 and SD levels compared with the negative control group. Similarly, rats treated with the combination of HYP EMLs, Ho-MNs and PDT showed superior joint healing efficacy compared with the groups treated with HYP EMLs in dark, HYP ointment or HYP in microneedles in histopathological examination. These findings highlight the promising potential of photoactivated HYP EMLs when combined with Ho-MNs technology for RA management. The presented therapeutic EMLs-MNs platform could serve as a powerful game-changer in the development of future localized RA treatments. [ABSTRACT FROM AUTHOR]

Abstract Background Colon cancer affects 1.23 million people worldwide and is the third most common malignant disease in men and the second in women. The only curative treatment is surgical resection, but a significant number of patients develop local recurrence or distant metastases. One of the alternative treatment methods for colon cancer is photodynamic therapy (PDT). In recent years, hypericin (HYP) derived from Hypericum perforatum has been suggested as a strong candidate photosensitizer for PDT. Our interest is focused on the biophysical changes in colon cancer cells in relation to HYP-mediated PDT. Results In this study, HYP-mediated PDT at 0.04, 0.08 or 0.15 μM HYP concentrations was performed in HT-29 colon adenocarcinoma cells and the Electron Paramagnetic Resonance (EPR) spectra of the spin labeled cells were obtained. Plasma membranes are already heterogeneous structures; the presence of cancer cells increased the heterogeneity and also fluidity of the plasma membranes. Therefore, the obtained spectra were evaluated by EPRSIMC program, which provides the calculation of heterogeneous structures up to four spectral components with different fluidity characteristics. Generally, two motional patterns were obtained from calculations and the number of them increased at the highest concentration. As the order parameters of the most populated components compared, an increase was observed depending on the HYP concentration. However, because of the heterogeneous structure of membrane, the order parameters of the less populated components did not exhibit a regular distribution. Conclusion After HYP-mediated PDT, concentration dependent changes were observed in the domain parameters indicating an increase in the HYP accumulation.

Abstract Background Adult T-cell leukemia (ATL) is an aggressive neoplasm caused by human T-cell leukemia virus type 1 (HTLV-1). ATL carries a poor prognosis due to chemotherapy resistance. Thus, it is urgent to develop new treatment strategies. Hypericin (HY) is a new-type of photosensitizer in the context of photodynamic therapy (PDT) due to its excellent photosensitizing properties and anti-tumor activities. Results In the present study, we investigated the efficacy of hypericin in ATL cells. Clinically achievable concentrations of hypericin in association with PDT induced the inhibition of cell proliferation in ATL cell lines with minimal effect on peripheral blood CD4+ T lymphocytes. Moreover, hypericin-PDT treatment caused apoptosis and G2/M phase cell cycle arrest in leukemic cells. Western blot analyses revealed that hypericin-PDT treatment resulted in downregulation of Bcl-2 and enhanced the expression of Bad, cytochrome C, and AIF. Cleavage of caspases-3/-7/-9/-8, Bid, and PARP was increased in hypericin-PDT-treated ATL cells. In a luciferase assay, hypericin-PDT treatment was able to activate the promoter activity of Bax and p53, resulting in enhanced expression of Bax and p53 proteins. Finally, hypericin-PDT treatment suppressed the expression of viral protein HBZ and Tax by blocking the promoter activity via HTLV-1 5′LTR and 3′LTR. Conclusions Our results revealed that hypericin-PDT is highly effective against ATL cells by induction of apoptosis and suppression of viral transcription. These studies highlight the promising use of hypericin-PDT as a targeted therapy for ATL.

Breast cancer resistance protein (BCRP) belongs to the family of ATP-binding cassette (ABC) transporters, overexpression of which can confer a multidrug-resistant phenotype in cancer cells and tumors. BCRP mediates efflux of numerous xenobiotics, including various chemotherapeutic agents and photosensitizers. Hypericin (HY) is a naturally-occurring photosensitizer synthesized by plants of the genus Hypericum. Our recently published results indicate that accumulation of HY in cancer cells of different tissue origin can be affected mostly by BCRP. Considering all known facts, the main goal of this study was to verify whether not only HY accumulation but also toxicity of HY-mediated photodynamic therapy (PDT) can be affected by the presence of some ABC transporters. To specifically prove our hypothesis, we used an experimental model of human leukemia cell lines differing in the expression level of the main drug efflux transporters P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1) and BCRP. The lowest HY accumulation, and consequently the highest resistance to HY-PDT, was found in cells overexpressing BCRP. Moreover, pretreatment with BCRP inhibitor Ko143 significantly increased HY accumulation and sensitized cells to HY-PDT. Therefore, our findings represent direct evidence that BCRP is the nemesis of HY accumulation and toxicity of HY-PDT. Thus, we should emphasize that individualized screening for BCRP expression and activity may represent a useful tool for prediction of HY-mediated photodynamic diagnosis (PDD) or PDT effectiveness.

Photodynamic therapy (PDT) is an established noninvasive tumor treatment. The hydrophobic natural occurring pigment hypericin shows a lot of attractive properties for the application in PDT. Hence, the administration to biological systems or patients requires the formulation in drug carriers enabling sufficient bioavailability. Therefore, free hypericin was encapsulated by the thin film hydration method or a hypericin-hydroxypropyl-β-cyclodextrin inclusion complex (Hyp-HPβCD) was incorporated by dehydration-rehydration vesicle method in either conventional or ultra-stable tetraether lipid (TEL) liposomes. The hydrodynamic diameter of the prepared nanoformulations ranged between 127 and 212 nm. These results were confirmed by atomic force microscopy. All liposomes showed a good stability under physiological conditions. TEL liposomes which tend to build more rigid bilayers, generate higher encapsulation efficiencies than their conventional counterparts. Furthermore, the suitability for intravenous application was confirmed by hemocompatibility studies resulting in a hemolytic potential less than 20% and a coagulation time less than 50 sec. The uptake of liposomal hypericin into human ovarian carcinoma cells (SK-OV-3) was confirmed using confocal microscopy and further characterized by pathway studies. It was demonstrated that the lipid composition and intraliposomal hypericin localization influenced the anti-vascular effect in the chorioallantoic membrane (CAM). While hypericin TEL liposomes exhibit substantial destruction of the microvasculature drug-in-cyclodextrin TEL liposomes showed no effect. Nevertheless, both formulations yielded severe photocytotoxicity in SK-OV-3 cells in a therapeutic dosage range. Conclusively, hypericin TEL liposomes would be perfectly suited for anti-vascular targeting while Hyp-HPβCD TEL liposomes could deliver the photosensitizer to the tumor site in a more protected manner.

The use of ultrasound (US) in combination with a responsive chemical agent (sonosensitizer) can selectively trigger the agent’s anticancer activity in a process called sonodynamic therapy (SDT). SDT shares some properties with photodynamic therapy (PDT), which has been clinically approved, but sets itself apart because of its use of US rather than light to achieve better tissue penetration. SDT provides anticancer effects mainly via the sonosensitizer-mediated generation of reactive oxygen species (ROS), although the precise nature of the underpinning mechanism is still under debate. This work investigates the SDT anticancer activity of hypericin (Hyp) in vitro in two- (2D) and three-dimensional (3D) HT-29 colon cancer models, and uses PDT as a yardstick due to its well-known Hyp phototoxicity. The cancer cell uptake and cellular localization of Hyp were investigated first to determine the proper noncytotoxic concentration and incubation time of Hyp for SDT. Furthermore, ROS production, cell proliferation, and cell death were evaluated after Hyp was exposed to US. Since cancer relapse and transporter-mediated multidrug resistance (MDR) are important causes of cancer treatment failure, the US-mediated ability of Hyp to elicit immunogenic cell death (ICD) and overcome MDR was also investigated. SDT showed strong ROS-mediated anticancer activity 48 h after treatment in both the HT-29 models. Specific damage-associated molecular patterns that are consistent with ICD, such as calreticulin (CRT) exposure and high-mobility group box 1 protein (HMGB1) release, were observed after SDT with Hyp. Moreover, the expression of the ABC transporter, P-glycoprotein (P-gp), in HT-29/MDR cells was not able to hinder cancer cell responsiveness to SDT with Hyp. This work reveals, for the first time, the US responsiveness of Hyp with significant anticancer activity being displayed, making it a full-fledged sonosensitizer for the SDT of cancer.

[Display omitted] • The coupling process is an effective strategy to prepare high purity products. • The total recovery ratio of hypericin was 80.6 % in the coupling process. • Hypericin with the purity of 98.7 % was obtained from the extracts efficiently. • XAD16 N column chromatography showed an excellent stability and reproducibility. Hypericin has shown promising potential for the prevention and treatment of cancer. Here we report facile strategy for the separation and purification of natural products from the extracts by demonstrating hypericin isolation from Hypericum perforatum L. extract using macroporous resin column chromatography (MRCC) followed by preparative liquid chromatography (PLC). Effect of various parameters including the kinds of resins, loading extract concentration, loading flow rate, types of eluent, and elution flow rate on the separation efficiency of MRCC for hypericin was investigated. The purity and purification ratio of hypericin was 71.8 % and 2.4 respectively, optimized by XAD16 N MRCC with a high stability and reproducibility. PLC parameters were determined by optimizing stationary phase, mobile phase, and injection sample amount. The high purity hypericin (98.7 %) was achieved by PLC and the total recovery ratio of hypericin was 80.6 % in the coupling process. [ABSTRACT FROM AUTHOR]

Objectives Hypericin is a polycyclic aromatic naphthodianthrone that occurs naturally. It is also an active ingredient in some species of the genus Hypericum. Emerging evidence suggests that hypericin has attracted great attention as a potential anticancer drug and exhibits remarkable antiproliferative effect upon irradiation on various tumour cells. This paper aims to summarise the anticancer effect and molecular mechanisms modulated by hypericin-medicated photodynamic therapy and its potential role in the cancer treatment. Key findings Hypericin-medicated photodynamic therapy could inhibit the proliferation of various tumour cells including bladder, colon, breast, cervical, glioma, leukaemia, hepatic, melanoma, lymphoma and lung cancers. The effect is primarily mediated by p38 mitogen-activated protein kinase (MAPK), JNK, PI3K, CCAAT-enhancer-binding protein homologous protein (CHOP)/TRIB3/Akt/mTOR, TRAIL/TRAIL-receptor, c-Met and Ephrin-Eph, the mitochondria and extrinsic signalling pathways. Furthermore, hypericin-medicated photodynamic therapy in conjunction with chemotherapeutic agents or targeted therapies is more effective in inhibiting the growth of tumour cells. Summary During the past few decades, the anticancer properties of photoactivated hypericin have been extensively investigated. Hypericin-medicated photodynamic therapy can modulate a variety of proteins and genes and exhibit a great potential to be used as a therapeutic agent for various types of cancer. [ABSTRACT FROM AUTHOR]

The well-known photosensitizers hypericin, harmane, and emodin are typical pigments of certain mushroom species—is this a coincidence or an indication towards a photoactivated defense mechanism in the phylum Basidiomycota? This perspective article explores this hypothesis by cross-linking the chemistry of fungal pigments with structural requirements from known photosensitizers and insights from photoactivated strategies in the kingdom Plantae. Thereby, light is shed on a yet unexplored playground dealing with ecological questions, photopharmaceutical opportunities, and biotechnological potentials. [ABSTRACT FROM AUTHOR]

Combined therapies are usually used to treat acne vulgaris since this approach can tackle various foci simultaneously. Using a combination of spectroscopic, computational, and microbiological techniques and methods, herein we report on the use of β-lactoglobulin as a double payload carrier of hypericin (an antimicrobial photodynamic agent) and all-trans retinoic acid (an anti-inflammatory drug) for S. aureus in vitro photodynamic inactivation. The addition of all-trans retinoic acid to hypericin-β-lactoglobulin complex renders a photochemically safe vehicle due to the photophysical quenching of hypericin, which recovers its photodynamic activity when in contact with bacteria. The ability of hypericin to photoinactivate S. aureus was not affected by retinoic acid. β-Lactoglobulin is a novel biocompatible and photochemically safe nanovehicle with strong potential for the treatment of acne.

Background: Hypericum is an important genus in the family Hypericaceae, which includes 484 species. This genus has been grown in temperate regions and used for treating wounds, eczema and burns. The aim of this study was to predict the content of hypericin in Hypericum perforatum in varied ecological and phenological conditions of habitat using artificial neural network techniques [MLP (Multi-Layer Perceptron), RBF (Radial Basis Function) and SVM (Support Vector Machine)]. Results: According to the results, the MLP model (R2 = 0.87) had an advantage over RBF (R2 = 0.8) and SVM (R2 = 0.54) models and it was relatively accurate in predicting hypericin content in H. perforatum based on the ecological conditions of site including soil types, its characteristics and plant phenological stages of habitat. The results of sensitivity analysis revealed that phenological stages, hill aspects, total nitrogen, altitude and organic carbon are the most influential factors that have an integral effect on the content of hypericin. Conclusions: The designed graphical user interface will help pharmacognosist, manufacturers and producers of medicinal plants and so on to run the MLP model on new data to easily discover the content of hypericin in H. perforatum by entering ecological conditions of site, soil characteristics and plant phenological stages. [ABSTRACT FROM AUTHOR]

Photoredox catalysis is a powerful and modern strategy for the synthesis of complex organic molecules. So far, this field has relied on the use of a limited range of metal-based chromophores or artificial organic dyes. Here, we show that the ubiquitous plant genus Hypericum can be used as an efficient photoredox catalyst. The dried flowers efficiently catalyze two typical photoredox reactions, a photoreduction and a photooxidation reaction, with a versatile substrate scope. Constitution analysis of the worldwide available plant genus indicated that naphthodianthrones, namely the compounds of the hypericin family, are crucial for the photocatalytic activity of the dried plant material. In situ UV-vis spectroelectrochemical methods provide insights into the mechanism of the photoreduction reaction where the radical dianion of hypericin (Hyp˙2−) is the catalytically active species. Our strategy provides a sustainable, efficient and an easy to handle alternative for a variety of visible light induced photocatalytic reactions. [ABSTRACT FROM AUTHOR]

• Ultrasonic and microwave field increase the extraction rate of hypericin. • The kinetic of hypericin extraction can be modeled by the Jander's diffusion model. • Values of kinetic parameters are lowest in microwave and highest in thermal field. • Activation of hypericin for extraction in all fields is produced via same mechanism. • Transferred quantized energy leads to δHCC/δHOC vibrations in hypericin molecule. The comparative kinetic analysis of total hypericin (TH) extraction from the pre-extracted flowers of Hypericum perforatum carried out under simultaneous external physical field and cooling reaction system operational conditions (SEPFC) was performed. The kinetics curves of TH extraction in SEPFC were measured in the temperature range from T = 298 to 328 K. It was proved that presence of ultrasonic and microwave field cause increasing of the extraction rate compared to conventional isothermal extraction. The possibility of describing the kinetics of TH extraction in SEPFC with empirical (Elovich's model) and physically-based (film-theory model) were explored. The kinetic complexity of the process was examined with the differential isoconversional method. By applying the model-fitting method the proper kinetic model of TH extraction in SEPFC was found: [1 − (1 − α)(1/3)]2 = k M · t. The values of the extraction rate constant (k M), activation energy (E a) and pre-exponential factor (ln A) of TH extraction were calculated. The effect of ultrasonic and microwave field on (a) k M ; (b) E a and (c) ln A of TH extraction was determined. The model mechanism of activation of hypericin molecule for extraction in SEPFC conditions was proposed. The values of wave number of resonant vibrations, vibration quantum number and anharmonicity parameter for the TH extraction are obtained. [ABSTRACT FROM AUTHOR]

Background: Nowadays, some studies have shown the effect of hypericin on cancer cells. However, considering the cytotoxicity of this plant and signs of anticancer activity in the plant, unfortunately, there is still no proper treatment for leukemia cancer cells. Therefore, the present study aims to evaluate the anticancer effect of hypericin in the treatment of leukemia cancer and its possible mechanism of action.Methods: In this study, the K562 cell line was treated with different concentrations of hypericin for 24 and 48 h. Detection of cell death was performed by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-tetrazolium bromide assay. The rate of cell apoptosis was measured by Annexin V/propidium iodide assay using flow cytometry. The expression of Bax, Bcl2, Myc, Mdm2, and P53 genes was evaluated by real-time polymerase chain reaction test, and immunocytochemistry (ICC) analysis was used for further evaluation of P53.Results: The results showed that hypericin has a dose-dependent cytotoxic effect on the K562 (in much less dose compared with cisplatin). According to flow cytometry results, cell apoptosis after exposure to hypericin for 24 h was 53%, and ICC analysis on p53 confirmed this. Furthermore, after 24 h of exposure to hypericin with IC50 concentration, the expression of P53 and Bax genes increased and the expression of the Bcl2, Myc, and Mdm2 gene decreased.Conclusion: The results showed that hypericin exerts its cytotoxicity on K562 cancer cells by downregulating Mdm2 and Myc. Based on the data acquired from the present study and many investigations till now, hypericin can be a good option for leukemia cancer cells treatment. [ABSTRACT FROM AUTHOR]

Thermal ablation has been commonly used as an effective treatment for hepatocellular carcinoma; however, peri-necrotic tumor residues after ablation play a significant role in tumor recurrence and poor prognosis. Therefore, developing agents that can effectively target and eliminate residual tumors is critically needed. Necrosis targeting strategies have potential implications for evaluating tumor necrosis areas and treating the surrounding residual tumors. To address this issue, we have developed a biodegradable nanoparticle with necrosis avidity that is compatible with fluorescence imaging, single photon emission computed tomography (SPECT) imaging, and necrosis targeted radiotherapy. The nanoparticles were synthesized using iodine-131-labeled hypericin ( 131 I-Hyp) as the core and amphiphilic copolymer poly(ethylene glycol)- block -poly(ε-caprolactone) (PEG-PCL) as the shell. The developed nanoparticle, PNP@( 131 I-Hyp), has a uniform spherical morphology with a size of 33.07 ± 3.94 and 45.93 ± 0.58 nm determined by cryogenic transmission electron microscopy (cryo-TEM) and dynamic light-scattering analysis (polydispersity index = 0.19 ± 0.01), respectively, and having a good stability and blood compatibility in vitro . In mouse subcutaneous ablated-residual tumor models, fluorescence and SPECT imaging demonstrated that PNP@( 131 I-Hyp) prominently accumulated in the tumor and was retained for as long as 168 h following intravenous injection. Moreover, ex vivo analyses showed that PNP@( 131 I-Hyp) mainly gathered in the necrotic zones of subcutaneous tumors and inhibited residual tumors by radiotherapy. In addition, histological examination of harvested organs and hematological analysis demonstrated that intravenous injection of 5 mCi/kg nanoparticles caused no gross abnormalities. This multifunctional nanoparticle, therefore, has necrosis imaging and targeted therapeutic effects on residual tumors after thermal ablation of hepatocellular carcinoma, showing potential for clinical application.

Hypericin was isolated from Diahyperon tincture for use as a standard sample. The chromatographic purity of the hypericin was confirmed by high-performance liquid chromatography/mass spectrometry. Hypericin has the only chromatographic peak with a retention time of 23.10 min and gave a characteristic signal for the molecular ion [M – H+]– with m/z 504.05 in the mass spectrum. Electronic absorption spectra of Diahyperon tincture and the MeOH extract of Hyperici herba (Hypericum perforatum) showed characteristic hypericin bands at 550 and 590 nm and a band at 665 nm that was due to the presence of chlorophyll. It was shown that excitation of hypericin in Diahyperon tincture at λex = 470 nm gave an emission spectrum with characteristic hypericin bands at 593 and 640 nm while the emission spectrum of the MeOH extract of Hyperici herba showed the hypericin emission bands and a weak chlorophyll emission band at 670 nm. It was found that spectrophotometric and spectrofluorimetric determinations of hypericin in Diahyperon tincture and in Hyperici herba gave similar results that were consistent with the standardized values of the hypericin content in the studied samples. [ABSTRACT FROM AUTHOR]

• We provide first detailed altitude-dependent chemical profiles of Hypericum perforatum populations • Content of polar metabolites increased with altitude of plant habitat • Altitude also changed essential oil composition significantly • Results indicate the investigated populations as new chemotypes in terms of their essential oil composition. • The findings could be useful to select unique strains with desired chemical composition Altitudinal variations in the content of main bioactive secondary metabolites namely, hypericin, pseudohypericin, hyperforin, the chlorogenic, neochlorogenic, caffeic and 2,4-dihydroxybenzoic acids, isoquercitrin, quercitrin, avicularin, hyperoside, rutin, biapigenin, (+)-catechin, (-)-epicatechin and in the composition of essential oil (EO) among Hypericum perforatum populations from two growing sites of Northern Turkey were investigated in detail for the first time. Aerial parts of 30 individuals at full flowering were collected at five different altitudes (391, 631, 1318, 1586 and 1733 m) of "Anzer" upland and six different altitudes (1311, 1447, 1968, 2068, 20169 and 2210 m) of "Cimil" upland in territory of Rize province, Turkey. Air dried plant samples were assayed for polar ingredients by high-performance liquid chromatography (HPLC) and for EO components by solid-phase microextraction (SPME) supported gas chromatography-mass spectrometry (GC-MS) analyses. All the tested polar phytochemicals were detected at various levels depending on altitude in both uplands and the accumulation level of each compound showed a positive consistent and significant response to altitudinal gradient. A considerable diversity in quality and quantity of major EO components was also observed and 2,2,6-trimethyl-cyclohexanone, caryophyllene oxide, tricycloheptane, hexanoic acid, tetradecanol, 2-methyl-dodecane, tetradecane, α-amorphene, eicosene, β-cedrene, piperitone, spathulenol, β-caryophyllene, δ-cadinene and linalool were identified as major EO components depending on altitude. The present results suggest that altitude of plant habitat has an ultimate effect on secondary metabolism of Hypericum perforatum and indicate the investigated populations as new chemotypes in terms of their EO composition. Such kind of findings could be useful to characterize the unique strains with desired chemical composition and can provide new viewpoints for investigations on this valuable medicinal plant. [ABSTRACT FROM AUTHOR]

The traditional medicinal herb, Hypericum perforatum L. has been popular for its pharmaceutical and coloring wealth since the ancient era. A secondary metabolite from the group of naphthodianthrones in Hypericum spp. named hypericin is responsible for the antidepression, anticancer, and antiviral characteristics of this herb. It has been found that several genes are involved in the biosynthesis pathway of hypericin. The hyp-1 gene is participating in this biosynthesis path through the conversion of emodin to hypericin. The naphthodianthrones (hypericin and pseudohypericin) in Hypericum are synthesized through the polyketide pathway. In the plants, the enzyme complexes named polyketide synthase (PKS) catalyzes the reactions of polyketide pathways. The genes HpPKS1 and HpPKS2 are encoding PKS enzyme complexes. In this research, the relative expression of hyp-1, HpPKS1, and HpPKS2 genes was compared in root and leaves of Hypericum perforatum and H. androsaemum L., before and after flowering under urea fertilization at 24, 48 and 72 hours after irrigation. The highest expression level of all three genes was observed after flowering in the samples of H. perforatum that were fertilized 72 hours after irrigation by 1 g l-1 urea (hyp-1 in roots; HpPKS1 and HpPKS2 in leaves). The relative expression of hyp-1 in the root was greater than in the leaves, but HpPKS1 and HpPKS2 expression in leaves was higher than in root. The relative expression of all three genes in H. perforatum was higher than in H. androsaemum. By increasing the interval between urea fertilization and irrigation, the relative expression of genes had an increasing trend, also by increasing the amount of urea fertilizer, relative gene expression was increased.

Background: Biotechnologic methods are common for secondary metabolites production from the plants and other sources in pharmaceutical sciences. Hairy root cell lines as the biotechnologic method have been used for in vitro production of major plant metabolites. Objective: In this study, hairy roots of Hypericum Perforatum have been prepared using the seeds and bio transformed by bacteria. Finally, the hypericin have been producted by the hairy roots. Methods: First, the seeds have been incubated in the plant media to hairy roots produced. Then, the hairy roots have been dipped in Rhizobium rizogenes suspension for biotransformation of bacterial genes. Morphological and phytochemical features of hairy roots have been determined in order to select the H. perforatum genotypes with higher hypericin contents. The fresh and dry weight of ten lines clones were measured after 30 days. In addition to, methanolic extracts of final hairy roots have been prepared and hypericin has been isolated and assayed by High Performance Liquid Chromatography. Results: According to the observations, one of ten hairy root lines shows 75-fold higher hypericin content (339.27 ppm) compared to non-transformed H. perforatum (4.58 ppm) in the same of body weight of roots. The clone with the most content of hypericin had significant development of biomass of hairy roots and increase the hypericin production. Transformed clones were varied in morphology, growth, and metabolite productivity. Conclusion: The mentioned methods induce the production of hairy root secondary metabolites in high scale to improve the quality and the quantity of pharmaceutical compounds.

Chen Shao, Kun Shang, Huaibao Xu, Yu Zhang, Zhichao Pei, Yuxin Pei Shaanxi Key Laboratory of Natural Products & Chemical Biology, College of Chemistry & Pharmacy, Northwest A&F University, Yangling, Shaanxi 712100, China Background: Photodynamic therapy is a safe, noninvasive modality for cancer therapy, in which the photosensitizer (PS) is a crucial component. Hypericin (Hy) is a promising PS; however, its clinical application is significantly limited by its poor hydrophilicity. Materials and methods: To overcome the clinical application limitation of Hy, a novel strategy is developed here by entrapping Hy into polydopamine (PDA) film formed on the surface of magnetic iron oxide nanoparticles (MNPs) through the self-polymerization of dopamine under alkaline condition. The amount of Hy in the Hy-entrapped PDA–MNP composite nanoparticles (denoted as PHMs) was measured by spectrophotometry. Furthermore, lactose, as the targeting ligand to asialoglycoprotein receptors, was conjugated to the surface of the PHMs by taking advantage of the spontaneous reaction of PDA with amino groups. Results: Spectrophotometry analysis revealed that the amount of Hy in the PHMs was 72 µmol g-1 PHMs. The fabricated Hy-entrapped glyconanoparticle (Lac-PHM) exhibited excellent water dispersibility, stability, and selectivity for asialoglycoprotein receptors overexpressing HepG2 cells. Atomic absorption spectroscopy analysis showed that the amount of the Lac-PHMs taken in HepG2 cells was 2.1-fold higher than that of the triethylene glycol-modified PHMs. The results of intracellular reactive oxygen species generation detection, cytotoxicity study, and apoptosis detection indicated that the Lac-PHMs had a satisfying photodynamic effect to HepG2 cells. Conclusion: The strategy developed in this work offers great potential for delivery of a variety of hydrophobic PSs. Keywords: hypericin, photodynamic therapy, glyconanoparticles, targeted drug delivery

INTRODUCTION: Inflammation is an innate immune response that protects host organism from external injuries and pathogens. The increased inflammatory response in organism can cause many diseases, especially cancer. Many natural and synthetic substances are used for annihilating the inflammatory response. Hypericin and hyperforin, isolated from Hypericum perforatum L., has shown anti-viral, anti-cancer, anti-depressive and anti-microbial properties. However, data on the mechanism of hypericin and hyperforin action against inflammation is limited. This study aimed at determining the anti-inflammatory property of hypericin and hyperforin using in vitro model. METHODS: In this study, 0, 25, 50, 75 and 100 nM hyperforin and 0, 2.5, 5, 7.5 and 10 µM hypericin were treated to lipopolysaccharide (LPS) stimulated RAW 264.7 macrophages. We purposed to display the anti-inflammatory effects of hypericin and hyperforin via measuring prostaglandin E2 (PGE2) and nitric oxide (NO) production and cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) gene expression in LPS induced RAW 264.7 macrophages. RESULTS: Our study has shown that all applied concentration of hyperforin and hypericin decreased nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) gene expression in RAW 264.7 macrophages, significantly. Similarly, all applied concentration of hyperforin and hypericin decreased prostaglandin E2 (PGE2) production but this decrease was statistically significant only in groups treated with 75 and 100 nM hyperforin and 7.5 and 10 μM hypericin. DISCUSSION AND CONCLUSION: This indicates that hyperforin and hypericin inhibited NO and PGE2 production and iNOS and COX-2 gene expression in RAW 264.7 macrophages in a dose-dependent manner. Inhibition of NO and PGE2 production by hyperforin and hypericin is a result of the inhibition of iNOS and COX-2 gene expression. For this reason, it is possible to say that hypericin and hyperforin can be used to reduce the inflammatory response.

SrY2O4 phosphors were doped with different concentrations of Eu3+ (0.5, 1, 2, 4, 8 and 10 at %) in order to investigate the maximal doping concentration of Eu3+ and its implementation in solar cell devices. Samples were synthesized by a combustion method using citric acid and glycine as a fuel. The X-ray diffraction (XRD) patterns confirmed pure phase of SrY2O4. FE-SEM micrographs showed agglomerate phenomenon with spherical-like shape particles and diameter of about 50 nm. Upon excitation with 280 nm, emission spectra were recorded in the range from 450-750 nm and in all samples the same energy transitions were observed 5D0 → 7FJ (J = 1, 2, 3 and 4) with maximal intensity for sample with 8 at % of Eu3+. That sample was further examined for the purpose of application in solar cell devices and showed high value of efficiency at low light intensities. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 45005]

Background and Objectives: The stress caused by maternal separation in the early days after birth induces autistic behaviors. Hypericin has many biological properties, including antidepressant and anti-inflammation. Therefore, this study aimed at determining hypericin effect on autism behaviors that induced by maternal separation model. Materials and Methods: In this experimental study, Wistar rats were divided into two control and maternal separation groups. The rats in the maternal separation group were subjected to individual separation from their mother for 1 h per day (postnatal day1-9). The maternal separation group was divided into 4 subgroups: Autism, fluoxetine, hypericin (10mg/kg), hypericin (50mg/kg). Treatment was gavaged in pups on postnatal day 21-34. In the end of day 34, neurobehavioral tests (grooming, social interaction and open field test) were carried out. Statistical analysis and between group comparison were performed using one-way ANOVA followed by Tukey's test. Results: In the pups subjected to individual isolations from their mother, social deficits (p=0.007), excessive repetitive behavior (p=0.001), and anxiety (p=0.001) were increased. Whereas, hypericin could reduce repetitive behaviors index (p=0.011), anxiety (p=0.001) and social interaction deficits (p=0.031) in the maternal isolation autism rat model. Conclusion: These findings indicated that maternal separation may produce autistic-like behaviors, and hypericin through antidepressant effect may be a potential therapeutic agent against autistic-like behavior. [ABSTRACT FROM AUTHOR]

Efficacy of photodynamic therapy (PDT) with hypericin was compared using two cell lines in monolayer culture. Photokilling was substantially impaired in the A549 (human‐derived lung tumor) when compared with the 1c1c7 murine hepatoma. The efficacy of PDT with regard to photokilling was correlated with the steady‐state intracellular hypericin concentration. There was otherwise no difference in subcellular targets or the pathway to cell death: paraptosis. In a prior study involving cell lines derived from patients with head and neck cancer, differences in response to PDT had shown no correlation with photosensitizer uptake. [ABSTRACT FROM AUTHOR]

Effects of photodynamic therapy (PDT) using the anthraquinone hypericin were explored with OVCAR‐5 cells in vitro. Irradiation resulted in ER > lysosomal photodamage. Paraptosis was identified as a primary death pathway resulting from ER perturbation. This is characterized by an extensive pattern of cytoplasmic vacuole formation. As the PDT dose increased, apoptotic death was also detected. The cytoprotective effect of autophagy, observed when certain other subcellular sites are PDT targets, appears to be absent. These results, together with prior evidence that paraptosis can be lethal to cells with an impaired apoptotic pathway, suggest a role for agents with this targeting profile in photodynamic therapy. A limitation to be overcome for hypericin is a suboptimal absorbance profile. [ABSTRACT FROM AUTHOR]

St. John's wort has been used for centuries in traditional medicine of many cultures, and nowadays it is well-known as a clinically important antidepressant drug. Considering the rising market demand for Hyperici herba, quality control of crude drug is of paramount importance. In this paper we performed HPLC-DAD chemical profiling of St. John's wort tea samples obtained at local markets, pharmacies and health food stores in the Balkan Peninsula countries, Austria and Turkey. Furthermore, water alcoholic extracts of the collected samples were evaluated in terms of their antioxidant potential, as well as the ability to inhibit biologically important enzymes such as acetylcholinesterase, monoamine oxidases A and B (MAO-A and MAO-B), α-amylase and α-glucosidase. Significant variability within the samples in the quantities of hypericin, hyperforin, rutin, quercetin, gallic, chlorogenic, caffeic and p-hydroxybenzoic acid was noticed. Chemotaxonomic modelling enabled the identification of three clusters of samples based on the quantities of rutin, hypericin and hyperforin. Generally, the extracts exhibited a significant potential to inhibit MAO-A (median IC50 = 10.01 μg/mL) and α-glucosidase (median IC50 = 12.40 μg/mL). The results of antioxidant potential evaluation suggest strong neutralization of hydroxyl and nitroso radicals, but moderate inhibition of lipid peroxidation process. Overall, the conducted study emphasizes the possibility of St. John's wort teas quality control, taking into account unavoidable variability of chemical profile as well. The stated is of great importance when grinding degree of herbal material excludes taxonomic identification of biological origin as an option within quality control. [ABSTRACT FROM AUTHOR]

It is of great importance to find an effective approach that not only eliminates gastric cancer cells but also do exhibits significant side effect to normal cells. Some studies have shown the effectiveness of hypericin against cancer cells. In this study, we evaluated the anti-cancer effect of Hypericin in the treatment of gastric cancer. In this study, the AGS cell line was exposed to different concentrations of hypericin for 24 and 48 h. Evaluation of cell death was done by MTT assay. The rate of apoptosis was measured by flow cytometry assay using Annexin V/ Propidium Iodide. The expression rate of Bcl2, p53 and Bax genes was evaluated by Real-time PCR test, and immunocytochemistry (ICC) analysis and western blotting was used for further evaluation of p53. MTT assay test showed that hyepricin induces 50% cell death in the concentration of 1 (μg/ mL) and 0.5 (μg/mL) at 24 h and 48 h post-treatment, respectively, however no similar effect seen on fibroblast cells. Annexin/PI test revealed that cell apoptosis after exposure to hypericin for 24 h was 74%. Real-time PCR showed that expression level of Bax, p53 and Bax genes increases and Bcl2 gene decreases in AGS cell lines after treatment by hypericin. ICC analysis and western blotting for p53 confirmed these data. The results of this study indicated that hypericin has the potential to be introduced as an effective treatment for gastric cancer. Therefore, it seems that this substance has potential to be utilized as anti-cancer drug. [ABSTRACT FROM AUTHOR]

Over the past decade, hypericin (Hy) has been used by several researchers due to its broad therapeutic potential against microorganisms and cancer cells. Considering the high hydrophobicity of Hy, its application in the biological medium becomes limited. In recent works, our research group has shown the potential of the liposomes to solubilize and carry Hy with high efficiency in different photodynamic therapy (PDT) applications. Moreover, previous works have shown that copolymer-coated liposomes have emerged as a smart alternative to conventional liposomes due to their targeting kinetic stability and prolonged circulation time in blood. About this approach, we report here a Hy delivery system based on the use of copolymer–lipid liposomal vesicles. We achieve a better proportion between the F127 copolymer and 1,2-dipalmitoyl-sn-3-glycerol-phosphatidylcholine by applying kinetic studies of association. The findings show that Hy was incorporated into the self-assembled copolymer–lipid with a t1/2 value five times lower, when compared to the conventional vesicles. The copolymer–lipid was subjected to cycles of thermal and temporal variations as well as to freeze-drying and reconstitution processes. We show that the F127 acts as a protective agent, keeping the copolymer–lipid/Hy stable for 6 months in the solid state, without the use of a cryoprotectant. The coated vesicles loading Hy showed high stability in the aqueous medium by 20 days, even when the temperature changed. Location studies by the fluorescence technique revealed specific interaction binding sites between the copolymer–lipid-coated and Hy molecules, showing that F127 plays an important role in the higher Hy solubilization. According to our results, we believe that this coated system is a potential Hy delivery system to PDT medical applications. [ABSTRACT FROM AUTHOR]

Hypericin (Hyp) is considered a promising photosensitizer for Photodynamic Therapy (PDT), due to its high hydrophobicity, affinity for cell membranes, low toxicity and high photooxidation activity. In this study, Hyp photophysical properties and photodynamic activity against melanoma B16-F10 cells were optimized using DPPC liposomes (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) as a drug delivery system. This nanoparticle is used as a cell membrane biomimetic model and solubilizes hydrophobic drugs. Hyp oxygen singlet lifetime (τ) in DPPC was approximately two-fold larger than that in P-123 micelles (Pluronic™ surfactants), reflecting a more hydrophobic environment provided by the DPPC liposome. On the other hand, singlet oxygen quantum yield values (ΦΔ1O2) in DPPC and P-123 were similar; Hyp molecules were preserved as monomers. The Hyp/DPPC liposome aqueous dispersion was stable during fluorescence emission and the liposome diameter remained stable for at least five days at 30 °C. However, the liposomes collapsed after the lyophilization/rehydration process, which was resolved by adding the lyoprotectant Trehalose to the liposome dispersion before lyophilization. Cell viability of the Hyp/DPPC formulation was assessed against healthy HaCat cells and high-metastatic melanoma B16-F10 cells. Hyp incorporated into the DPPC carrier presented a higher selectivity index than the Hyp sample previously solubilized in ethanol under the illumination effect. Moreover, the IC50 was lower for Hyp in DPPC than for Hyp pre-solubilized in ethanol. These results indicate the potential of the formulation of Hyp/DPPC for future biomedical applications in PDT treatment. [ABSTRACT FROM AUTHOR]

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Hypericin (Hyp), a natural hydrophobic and photoactive pigment, and methylene blue (MB), a hydrophilic cationic dye, are utilized as photosensitizer (PS) for photodynamic therapy of cancer. Bioadhesive and thermoresponsive polymeric systems can improve the drug availability by increasing the contact time between the system and the mucosa and also controlling the drug release. In this work, an accelerated physicochemical stability study of binary polymeric systems composed of poloxamer 407 (Polox) and Carbopol 934 P (Carb) for MB or Hyp release was performed. Formulations were prepared containing Polox (20%, w/w), Carb (0.15%, w/w) and MB (0.25%, w/w) or Hyp (0.01%, W/W) and submitted to different stress conditions (5 ± 3 °C, 25 ± 2 °C and 40 ± 2 °C with relative humidity of 75 ± 5%) during 180 days. The samples were analyzed as macroscopic characteristics, photosensitizer content and mechanical properties by texture profile analysis. Both systems displayed decrease of photosensitizer content less than 5% during 180 days. MB-system showed an undefined reaction model, while Hyp-system displayed PS decay following a pseudo first-order reaction. Systems also displayed stable mechanical characteristics. The pharmaceutical analyses showed the good physicochemical stability of the bioadhesive platform for delivery Hyp and MB in photodynamic therapy. [ABSTRACT FROM AUTHOR]

Herbal medicines are frequently used for the prevention and treatment of obesity and obesity-related disorders. Our preliminary screening showed that St. John's Wort (SJW) displayed potent inhibition on pancreatic lipase (PL), a key hydrolase responsible for lipid digestion and absorption in mammals. Herein, the inhibition potentials and inhibitory mechanism of SJW extract and its major constituents on PL were fully investigated by a set of in vitro and in silico studies. The results clearly demonstrated that the naphthodianthrones, biflavones and most of flavonoids in SJW displayed strong to moderate inhibition on PL. Among all tested natural compounds, two naphthodianthrones (hypericin and pseudohypericin) and one biflavone (I3,II8-biapigenin) isolated from SJW exhibited potent PL inhibition activity, with the IC 50 values of <1 μM. Inhibition kinetics analyses showed that hypericin, pseudohypericin and I3,II8-biapigenin inhibited PL via a mixed manner, while molecular dynamics simulations revealed that three newly identified PL inhibitors could bind on PL at both the catalytic cavity and the interface between colipase and the C-terminal domain of PL. Collectively, our findings suggested that part of major constituents in SJW displayed potent PL inhibition activities, which could be used as lead compounds for the development of novel PL inhibitors. • The inhibitory effects of St. John's wort extract and its major constituents on pancreatic lipase (PL) are assayed. • Hypericin, pseudohypericin and I3,II8-biapigenin display potent inhibition on PL, with the IC 50 values of <1 μM. • Hypericin, pseudohypericin and I3,II8-biapigenin are mixed inhibitors of PL. • Hypericin, pseudohypericin and I3,II8-biapigeninc could bind on PL at two distinct ligand-binding sites. [ABSTRACT FROM AUTHOR]

The G‐quadruplex (G4)/hemin entity has been widely used as a peroxidase‐mimic DNAzyme to catalyze the substrate oxidation with exogenous and concentrated H2O2 as oxidant for developing various devices. Herein, a G4‐based oxidase‐mimic DNAzyme that can be driven by visible light was developed. The oxidant is in situ produced by light irradiation. As a natural photosensitizer, Hypericin (Hyp) alone in solution is inactive because of aggregation. However, the binding with G4 triggers the fluorescence emission and activates Hyp to produce singlet oxygen (1O2) by energy transfer to the dissolved oxygen. Therefore, the G4/Hyp entity can serve as a photooxidase via the 1O2 pathway to catalyze the substrate oxidation. Since the G4‐bound Hyp can be excited by almost the whole range of visible light, the photooxidase should cover a wide range of photocatalytic applications. Similar to the peroxidase‐mimic DNAzyme, the G4 sequence can regulate the photooxidase activity and metal ions can serve as efficient cofactors to activate the photooxidase capacity for an inactive G4 structure. This work provides an alternative to build a versatile G4‐based photooxidase with straightforward tunability of its activity. [ABSTRACT FROM AUTHOR]

Majority of synthetic drugs used by contemporary medicine originate from substances present in plants. However, natural sources of herbal drugs are being erratic, cultivated medicinal plants also face climate change, whereas the demand for bioactive compounds and plant extracts is on the increase. Rational phytotherapy applies preparations with well identified active components, whose pharmacological efficiency has been proved by clinical tests. To perform the studies in vivo using animals and for clinical tests significant amounts of bioactive compounds are required. The compounds and plants of interest include menthol, main component of Peppermint oil, salicin from Willow bark and 1-deoxynojirimycin from Morus alba. These compounds can be isolated in small amounts from herbs, but the methods of their total organic synthesis should be developed. It is an inspiration for chemists and first step towards industrial production. [ABSTRACT FROM AUTHOR]

• Hypericum aviculariifolium and H. pruinatum were cultured in vitro for the first time. • Elicitation of the cultures with jasmonic acid accelerated phytochemical accumulation. • The results offer a strategy for conservation of these endemic and rare species, and could be used for enhancing the production of certain bioactive phytochemicals. The aim of the study was to introduce two endangered species from Turkish flora, Hypericum aviculariifolium and H. pruinatum , into in vitro culture and to determine the elicitation effect of jasmonic acid (JA) on the accumulation of secondary metabolites in vitro regenerated flowering plants. A Murashige and Skoog (MS) medium supplemented with plant growth regulators were used for seed germination and seedling culture. Seedling shoot explants in MS medium supplemented with combination of 1 mg/l BA and IAA resulted in maximum and simultaneous induction of roots and shoots. The regenerants of both species survived (98%) in greenhouse cultivation and were assayed for bioactive compounds. The content of secondary metabolites of flowering plants, originating from in vitro culture and treated with different doses of JA was higher than that of intact, i.e. maternal plants providing seeds and control plants. However, the intact and control plants did not differ significantly in the amounts of the bioactive compounds tested. The chemical profile of both species was similar with quantitative differences of individual compounds. Successful establishment of a highly efficient in vitro multiplication protocol in the present study offers a crucial in vitro strategy for the further conservation of endemic and rare species and enhanced production of valuable secondary metabolites. [ABSTRACT FROM AUTHOR]

The aim of this study was to measure the secretion of interleukin (IL)-8 and -10 during an elicited immune response following sublethal doses of hypericin-mediated photodynamic therapy (HY-PDT) in experimental models of residual colon cancer cells in vitro. Investigations were performed on the cancer cell lines SW480 and SW620. Each cell line was exposed to 3 different concentrations of the photosensitizer HY and various doses of irradiation. The cell metabolic activity using an MTT assay was performed and then the measurement of IL-8 and IL-10 secretion was achieved using the Bio-Plex ProTMAssay. There was a statistically significant amplification of IL-8 secretion during HY-PDT in the SW620 cell line (at 1 J/cm2: P =.01, 5 J/cm2: P =.002, and 10 J/cm2: P =.025) and a statistically significant decrease in IL-8 during HY-PDT in the SW480 cell line (at 1 J/cm2: P =.05, 5 J/cm2: P =.035, and 10 J/cm2: P =.035). No statistically significant differences in IL-10 concentration were found following HY-PDT in the SW480 (at 1 J/cm2: P >.4, 5 J/cm2: P =.1, and 10 J/cm2: P =.075) or in the SW620 cell line (at 1 J/cm2: P >.4, 5 J/cm2: P >.4, and 10 J/cm2: P >.4). HY-PDT can both eliminate and control a primary tumor via cytotoxic effects, and at sublethal doses, it can affect IL release by colon cancer cells. In this experiment, this influence depended on the level of tumor cell metastatic activity. [ABSTRACT FROM AUTHOR]

Summary: Hypericin is a molecule of high pharmaceutical importance that is synthesized and stored in dark glands (DGs) of St. John's Wort (Hypericum perforatum). Understanding which genes are involved in dark gland development and hypericin biosynthesis is important for the development of new Hypericum extracts that are highly demanded for medical applications. We identified two transcription factors whose expression is strictly synchronized with the differentiation of DGs. We correlated the content of hypericin, pseudohypericin, endocrocin, skyrin glycosides and several flavonoids with gene expression and DG development to obtain a revised model for hypericin biosynthesis. Here, we report for the first time genotypes which are polymorphic for the presence/total absence (G+/G−) of DGs in their placental tissues (PTs). DG development was characterized in PTs using several microscopy techniques. Fourier transform infrared microscopy was established as a novel method to precisely locate polyaromatic compounds, such as hypericin, in plant tissues. In addition, we obtained transcriptome and metabolome profiles of unprecedented resolution in Hypericum. This study addresses for the first time the development of dark glands and identifies genes that constitute strong building blocks for the further elucidation of hypericin synthesis, its manipulation in plants, its engineering in microbial systems and its applications in medical research. [ABSTRACT FROM AUTHOR]

Avian infectious bronchitis virus (IBV), a coronavirus, causes infectious bronchitis leading to enormous economic loss in the poultry industry worldwide. Hypericin (HY) is an excellent compound that has been investigated in antiviral, antineoplastic, and antidepressant. To investigate the inhibition effect of HY on IBV infection in chicken embryo kidney (CEK) cells, 3 different experimental designs: pre-treatment of cells prior to IBV infection, direct treatment of IBV-infected cells, and pre-treatment of IBV prior to cell infection were used. Quantitative real-time PCR (qRT-PCR), immunofluorescence assay (IFA), flow cytometry, and fluorescence microscopy were performed and virus titer was determined by TCID50. The results revealed that HY had a good anti-IBV effect when HY directly treated the IBV-infected cells, and virus infectivity decreased in a dose-dependent manner. Furthermore, HY inhibited IBV-induced apoptosis in CEK cells, and significantly reduced the mRNA expression levels of Fas, FasL, JNK, Bax, Caspase 3, and Caspase 8, and significantly increased Bcl-2 mRNA expression level in CEK cells. In addition, HY treatment could decrease IBV-induced reactive oxygen species (ROS) generation in CEK cells. These results suggested that HY showed potential antiviral activities against IBV infection involving the inhibition of apoptosis and ROS generation in CEK cells. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) is an established noninvasive tumor treatment. The hydrophobic natural occurring pigment hypericin shows a lot of attractive properties for the application in PDT. Hence, the administration to biological systems or patients requires the formulation in drug carriers enabling sufficient bioavailability. Therefore, free hypericin was encapsulated by the thin film hydration method or a hypericin-hydroxypropyl-β-cyclodextrin inclusion complex (Hyp-HPβCD) was incorporated by dehydration-rehydration vesicle method in either conventional or ultra-stable tetraether lipid (TEL) liposomes. The hydrodynamic diameter of the prepared nanoformulations ranged between 127 and 212 nm. These results were confirmed by atomic force microscopy. All liposomes showed a good stability under physiological conditions. TEL liposomes which tend to build more rigid bilayers, generate higher encapsulation efficiencies than their conventional counterparts. Furthermore, the suitability for intravenous application was confirmed by hemocompatibility studies resulting in a hemolytic potential less than 20% and a coagulation time less than 50 sec. The uptake of liposomal hypericin into human ovarian carcinoma cells (SK-OV-3) was confirmed using confocal microscopy and further characterized by pathway studies. It was demonstrated that the lipid composition and intraliposomal hypericin localization influenced the anti-vascular effect in the chorioallantoic membrane (CAM). While hypericin TEL liposomes exhibit substantial destruction of the microvasculature drug-in-cyclodextrin TEL liposomes showed no effect. Nevertheless, both formulations yielded severe photocytotoxicity in SK-OV-3 cells in a therapeutic dosage range. Conclusively, hypericin TEL liposomes would be perfectly suited for anti-vascular targeting while Hyp-HPβCD TEL liposomes could deliver the photosensitizer to the tumor site in a more protected manner. [ABSTRACT FROM AUTHOR]

Hypericin, a major bioactive compound from St. John's-wort, has been shown to exert several pharmacological activities including anti-viral, anti-depressant, and anti-cancer effects. However, the effect of hypericin on embryonic development still remains unclear. In this study, we investigated the toxicity of hypericin during embryonic development by using zebrafish as a model organism. Hypericin caused mortality and malformation in zebrafish in a dose- and time-dependent manner. In addition, hypericin also affected the liver development in zebrafish. On the other hand, no overt alteration in angiogenesis was observed in hypericin-treated zebrafish. In addition, no inhibition of Hep3B cell proliferation was observed using xenotransplantation assay in response to hypericin treatment. In conclusion, our findings suggest that hypericin may exhibit hepatotoxicity and lead to malformation and mortality during embryonic development in zebrafish. [ABSTRACT FROM AUTHOR]

Bacteremia is a life-threating syndrome often caused by methicillin-resistant Staphylococcus aureus (MRSA). Thus, there is an urgent need to develop novel approaches to successfully treat this infection. Staphylococcal accessory regulator A (SarA), a global virulence regulator, plays a critical role in pathogenesis and β -lactam antibiotic resistance in Staphylococcus aureus. Hypericin is believed to act as an antibiotic, antidepressant, antiviral and non-specific kinase inhibitor. In the current study, we investigated the impact of hypericin on β -lactam antibiotics susceptibility and mechanism(s) of its activity. We demonstrated that hypericin significantly decreased the minimum inhibitory concentrations of β -lactam antibiotics (e. g., oxacillin, cefazolin and nafcillin), biofilm formation and fibronectin binding in MRSA strain JE2. In addition, hypericin significantly reduced sarA expression, and subsequently decreased mecA, and virulence-related regulators (e. g., agr RNAⅢ) and genes (e. g., fnbA and hla) expression in the studied MRSA strain. Importantly, the in vitro synergistic effect of hypericin with β -lactam antibiotic (e. g., oxacillin) translated into in vivo therapeutic outcome in a murine MRSA bacteremia model. These findings suggest that hypericin plays an important role in abrogation of β -lactam resistance against MRSA through sarA inhibition, and may allow us to repurpose the use of β -lactam antibiotics, which are normally ineffective in the treatment of MRSA infections (e. g., oxacillin). In this study, the authors demonstrated that hypericin, as a sarA inhibitor, had synergistic activity with β -lactams both in vitro and in a murine bacteremia model due to MRSA. The mechanisms may be related to significantly reduced biofilm formation, fibronectin binding and expression of virulence genes (e. g., fnbA and hla). Image 1 [ABSTRACT FROM AUTHOR]

• Glioblastoma spheroids were grown in hanging drops to develop cell. • Fluorescence microscopy was used to examine the degree of heterogeneity in the spheroids. • Hypericin was used to induce photodynamic therapy in spheroids. • Autophagy and apoptosis were found to be induced in the periphery of spheroids. In recent years, pharmacology and toxicology have emphasised the intention to move from in vivo models to simplified 3D objects represented by spheroidal models of cancer. Mitochondria are one of the subcellular organelles responsible for cell metabolism and are often a lucrative target for cancer treatment including photodynamic therapy (PDT). Hanging droplet-grown glioblastoma cells were forced to form spheroids with heterogeneous environments that were characterised by fluorescence microscopy and flow cytometry using fluorescent probes sensitive to oxidative stress and apoptosis. PDT was induced with hypericin at 590 nm. It was found that the metabolic activity of the cells in the periphery and core of the spheroid was different. Higher oxidative stress and induction of caspase-3 were observed in the peripheral layers after PDT. These parts were more destabilised and showed higher expression of LC3B, an autophagic marker. However, the response of the whole system to the treatment was controlled by the cells in the core of the spheroids, which were hardly affected by the treatment. It has been shown that the depth of penetration of hypericin into this system is an important limiting step for PDT and the induction of autophagy and apoptosis. In this work, we have described the fluorescence imaging of vital mitochondria, caspase-3 production and immunostaining of autophagic LC3B in cells from glioblastoma spheroids before and after PDT. Overall, we can conclude that this model represents an in vitro and in vivo applicable alternative for the study of PDT in solid microtumours. [Display omitted] [ABSTRACT FROM AUTHOR]