Selective photodynamic effects on cervical adenocarcinoma cells provided by F127 Pluronic®-based micelles modulating hypericin delivery

Abstract Cervical cancer is a leading cause of death among women. The endocervical adenocarcinoma (ECA) represents an aggressive and metastatic type of cancer with no effective treatment options currently available. We evaluated the antitumoral and anti-migratory effects of hypericin (HYP) encapsulated on Pluronic F127 (F127/HYP) photodynamic therapy (PDT) against a human cell line derived from invasive cervical adenocarcinoma (HeLa) compared to a human epithelial cell line (HaCaT). The phototoxicity and cytotoxicity of F127/HYP were evaluated by the following assays: colorimetric assay, MTT, cellular morphological changes by microscopy and long-term cytotoxicity by clonogenic assay. In addition, we performed fluorescence microscopy to analyze cell uptake and subcellular distribution of F127/HYP, cell death pathway and reactive oxygen species (ROS) production. The PDT mechanism was determined with sodium azide and D-mannitol and cell migration by wound-healing assay. The treatment with F127/HYP promoted a phototoxic result in the HeLa cells in a dose-dependent and selective form. Internalization of F127/HYP was observed mainly in the mitochondria, causing cell death by necrosis and ROS production especially by the type II PDT mechanism. Furthermore, F127/HYP reduced the long-term proliferation and migration capacity of HeLa cells. Overall, our results indicate a potentially application of F127/HYP micelles as a novel approach for PDT with HYP delivery to more specifically treat ECA.

Research progress on the role of transient receptor potential canonical 6 in ischemia-reperfusion injury / 器官移植

Ischemia-reperfusion injury (IRI) refers to the reperfusion injury caused by the recovery of blood supply of ischemic tissues or organs, which commonly occurs in organ transplantation and other surgical procedures. IRI may cause a series of severe clinical issues, such as delayed graft function, acute kidney injury, myocardial infarction, ischemic stroke and circulatory arrest, etc. These events yield high incidence and fatality. At present, no effective solution has been available. Transient receptor potential canonical 6 (TRPC6), a member of Ca2+ channel family, is highly expressed in multiple types of cells. It may adjust many physiological functions by regulating intracellular Ca2+ concentration, which has become an important target for developing therapeutic drugs for multiple diseases. In this article, research progresses on the introduction and function of TRPC6, the association between TRPC6 and IRI and the therapeutic prospect of TRPC6 targeted drugs in IRI were reviewed, aiming to provide novel insights into the prevention and treatment of IRI during organ transplantation

Anti-liver Cancer Mechanism of Hypericin Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of hypericin against liver cancer using network pharmacology. MethodThe traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP）， Drug Gene Interaction Database （DGIdb）， Comparative Toxicogenomics Database （CTD） and SwissTargetPrediction were used to predict the targets of hypericin. Five databases including GeneCards and Online Mendelian Inheritance in Man （OMIM） were employed to obtain liver cancer-related targets. The intersection was performed to obtain the targets of hypericin against liver cancer. The Database for Annotation， Visualization and Integrated Discovery （DAVID） v2021q4 was used for Gene Ontology （GO） function annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. The protein-protein interaction （PPI） network of the targets was constructed by Cytoscape 3.6.1 to screen the core targets，and the affinity between hypericin and the core targets was verified by molecular docking. The effects of hypericin on liver cancer and the migration of liver cancer cells were observed by cell viability assay and would healing assay， respectively， and its effects on the mRNA and protein expression of key targets cysteinyl aspartate-specific protease-3（Caspase-3） and mitogen-activated protein kinase 3 （MAPK3） were detected by real-time polymerase chain reaction（Real-time PCR） and Western blot， respectively. ResultA total of 45 genes related to the anti-liver cancer effect of hypericin were obtained， and six core target genes were screened. The signal pathways enriched by KEGG pathway analysis included apoptosis，tumor necrosis factor （TNF） and cancer signal pathways. Molecular docking showed that the core target genes Caspase-3，TNF，estrogen receptor 1 （ESR1），MAPK3，catalase （CAT） and cyclooxygenase 2 （PTGS2） had good affinity with hypericin，especially Caspase-3 and MAPK3. In addition，compared with the conditions in control group， cell experiments demonstrated that hypericin could reduce the viability of liver cancer cells （P<0.05），inhibit their migration，increase the mRNA expression of Caspase-3 （P<0.05） and decrease that of MAPK3 （P<0.05）. ConclusionHypericin exerted the anti-liver cancer effect by affecting the core targets such as Caspase-3，TNF，ESR1，MAPK3，CAT and PTGS2 and jointly interfering with apoptosis，TNF and cancer signal pathways.

Hypericin enhances -lactam antibiotics activity by inhibiting expression in methicillin-resistant

Bacteremia is a life-threating syndrome often caused by methicillin-resistant (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 -lactam antibiotic resistance in . 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 -lactam antibiotics susceptibility and mechanism(s) of its activity. We demonstrated that hypericin significantly decreased the minimum inhibitory concentrations of -lactam antibiotics (.., oxacillin, cefazolin and nafcillin), biofilm formation and fibronectin binding in MRSA strain JE2. In addition, hypericin significantly reduced expression, and subsequently decreased and virulence-related regulators (.., ) and genes (.., and ) expression in the studied MRSA strain. Importantly, the synergistic effect of hypericin with -lactam antibiotic (.., oxacillin) translated into therapeutic outcome in a murine MRSA bacteremia model. These findings suggest that hypericin plays an important role in abrogation of -lactam resistance against MRSA through inhibition, and may allow us to repurpose the use of -lactam antibiotics, which are normally ineffective in the treatment of MRSA infections (.., oxacillin).

Mechanism of Photodynamic Therapy Mediated by Hypericin in Inhibiting the Proliferation of Leukemia Cell Lines / 中国药学杂志

OBJECTIVE: To observe the effects of hypericin on proliferation, apoptosis of leukemia cells and its possible mechanism. METHODS: Subcellular localization of hypericin in leukemia cells by fluorescence microscopy. MTT(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) test was adopted to observe the inhibitory effects of hypericin on the proliferation of leukemia cells and calculated its half maximal inhibitory concentration(IC50). The technology of flow cytometry, Annexin-V-FITC/PI double stained method were employed to measure the cell apoptosis of leukemia cells after hypericin treatment. RESULTS: The optimal time for hypericin to entry into cells was 16 h. Hypericin could inhibit the proliferation of leukemia cells in vitro, and the inhibition presented obvious dose-effect relationship(r=0.990 5, P<0.01). The apoptotic ratio of leukemia cells was gradually increased. The ratio of Bax/Bcl-2 was significantly increased, and the apoptotic protein caspase-3, 8, 9 is cleaved and activated. CONCLUSION: Hypericin could inhibit the growth of leukemia cells, induce the apoptosis through induction of caspase dependent apoptosis pathway.

Topically applied hypericin exhibits skin penetrability on nude mice / 中国药理学与毒理学杂志

OBJECTIVE Hypericin, a powerful naturally photosensitizer in photodynamic therapy (PDT), is suitable for treating skin diseases involving excess capillary proliferation. In the present study, we aimed to evaluate the skin penetrability of a topically applied hypericin, expecting reducing the risk of prolonged skin photosensitivity, which often occurs after systemic administration. METHODS The Franz diffusion cell assay was performed to evaluate different penetration enhancers. In vivo studies, fluorescence microscopy was performed to examine the distribution of hypericin in the skin, macroscopic and microscopic analyses were also carried out to detect pathological changes in the skin after topical hypericin-PDT treatment. Immunohistochemistry was used to determine the expression of PECAM-1 in the treated skin. RESULTS 5% menthol facilitated hypericin penetrate the skin of nude mice most. The results of in vivo assays revealed that hypericin penetrated nude mice skin, spread to the dermis, and resulted in obvious photosensitivity reaction on the dermal capillaries. Moreover, skin injured by the photosensitive reaction induced by hypericin was replaced by normal skin 7 d after hypericin-PDT treat?ment. CONCLUSION Topical hypericin could penetrate nude mouse skin well and be great potential in PDT treatment of skin diseases.

In vitro study on anti-HBV effects and mechanism of hypericin / 重庆医学

Objective To evaluate the anti-HBV effect of hypericin from the cellular level and to preliminarily explore its po-tential drug target point.Methods Liver cell line HepG2.2.15 cells secreting HBV particles were selected as the experimental ob-jects.Hypericin served as the HY group,lamivudine was taken as 3TC group and deionized water as the blank control group.The cells were grouped and administrated.The HBV-DNA copy level was measured at72 h after medication by Southern blot and fluo-rescent quantitative PCR;the inhibition rate of HBsAg and HBeAg was detected by using ELISA assay;the pgRNA expression level was tested by using Northern blot and fluorescent quantitative PCR;Western blot and fluorescent quantitative PCR were adopted to detect the expression of regulatory factors including HNF3β,HNF4α,PPARαand RXRα.Results Compared to the blank control group,both hypericin and lamivudine had significant inhibiting effect on HBV DNA and expression level of HBsAg and HBeAg in HepG2.2.15 cells (P <0.05).Hypericin could significantly decrease the pgRNA expression compared with the blank control group (P <0.05),while lamivudine had no obvious change (P <0.05).Moreover,hypericin exhibited significant effects on the expression of HNF3βand regulatory factor HNF4αcompared with the blank control group and 3TC group(P <0.05).Conclusion Hypericin represents a strong anti-HBV effect,moreover could increase the negative regulatory factor HNF3βn expression and decreases the positive factor HNF4αexpression,prompting that its drug target point could be pgRNA.

Hypericin, a Naphthodianthrone Derivative, Prevents Methylglyoxal-Induced Human Endothelial Cell Dysfunction

Methylglyoxal (MGO) is a highly reactive metabolite of glucose which is known to cause damage and induce apoptosis in endothelial cells. Endothelial cell damage is implicated in the progression of diabetes-associated complications and atherosclerosis. Hypericin, a naphthodianthrone isolated from Hypericum perforatum L. (St. John’s Wort), is a potent and selective inhibitor of protein kinase C and is reported to reduce neuropathic pain. In this work, we investigated the protective effect of hypericin on MGO-induced apoptosis in human umbilical vein endothelial cells (HUVECs). Hypericin showed significant anti-apoptotic activity in MGO-treated HUVECs. Pretreatment with hypericin significantly inhibited MGO-induced changes in cell morphology, cell death, and production of intracellular reactive oxygen species. Hypericin prevented MGO-induced apoptosis in HUVECs by increasing Bcl-2 expression and decreasing Bax expression. MGO was found to activate mitogen-activated protein kinases (MAPKs). Pretreatment with hypericin strongly inhibited the activation of MAPKs, including P38, JNK, and ERK1/2. Interestingly, hypericin also inhibited the formation of AGEs. These findings suggest that hypericin may be an effective regulator of MGO-induced apoptosis. In conclusion, hypericin downregulated the formation of AGEs and ameliorated MGO-induced dysfunction in human endothelial cells.

Metabolites of hypericin in rats in vivo / 中国药科大学学报

@#The study aimed to separate and identify the metabolites of hypericin in the bile and necrotic tissues in rats. After intravenous injection of 10 mg/kg hypericin, 0-12 h bile of normal rats and 24 h necrotic liver of rats with reperfused hepatic infarction were collected, and metabolites of rats were analyzed by high performance liquid chromatography coupled with electrospray tandemtime of flight mass spectrometry(HPLC-TOF/MS). The prototype(M0)and three glycosylation metabolites(M1, M2, M3)of hypericin in rat bile and the parent compound in rat necrotic liver were detected and identified. Results indicated that prototype and glycosylation of hypericin were the major metabolic form in rat bile and the parent compound was found only in necrotic tissues.

The tissues distribution of hypericin in rats / 中国药学杂志

OBJECTIVE: To study the tissues distribution of hypericin in rats. METHODS: A single dose of 50 mg·kg-1 hypericin was administered to healthy SD rats via tail vein. The tissues of heart, liver, spleen, lung, kidney, stomaeh and small intestine were acquired at 20, 45 or 90 min after administration. The concentrations of hypericin in tissues samples were determined by HPLC. RESULTS: After a single tail vein dose of 50 mg·kg-1 hypericin in rats, hypericin was mainly distributed in kidney. The concentrations of hypericin in heart was low. CONCLUSION: This present investigation on hypericin tissues distribution provides important information to further study and clinic use of hypericin.

Aspectos botânicos, químicos, farmacológicos e terapêuticos do Hypericum perforatum L / Botanical, chemical, pharmacological and therapeutic aspects of Hypericum perforatum L

Estima-se que aproximadamente 25% das drogas prescritas em todo o mundo são oriundas de espécies vegetais. Dentre as plantas com alto potencial medicinal, se destaca o Hypericum perforatum L. (HP), planta herbácea perene, pertencente à família Hypericaceae. Extratos orgânicos e aquosos de HP têm sido utilizados na medicina popular e em testes pré-clínicos para o tratamento e prevenção de diversas doenças através de efeitos nefroprotetores, atividades antioxidante, antifúngica, ansiolítica, antiviral e cicatrizante. Estudos clínicos indicaram que esta espécie pode ser útil no tratamento de desordens originadas do sistema nervoso central, especialmente na depressão unipolar. HP contém, ao menos, dez classes de compostos biologicamente ativos, dentre eles antraquinonas/naftodiantronas, derivados de floroglucinol, flavonoides, biflavonas, xantonas, óleos voláteis, aminoácidos, vitamina C, cumarinas, taninos e carotenoides. Ao mesmo tempo em que os constituintes possuem relevantes efeitos farmacológicos, os mesmos podem prejudicar, por antagonismo farmacocinético (interação com algumas enzimas do citocromo), a eficácia de outros fármacos. Devido a relevante importância do HP como agente terapêutico, ressalta-se a importância do desenvolvimento de novos estudos com o intuito de elucidar questões ainda controversas acerca do extrato de HP, e.g., dose, melhor horário para colheita, padronização dos extratos, e possíveis efeitos tóxicos, podendo assim, definir claramente os riscos e benefícios da utilização desta planta.

It is estimated that approximately 25% of prescribed drugs are derived from plant species. Among the plants with high medicinal potential, it highlights the Hypericum perforatum L. (HP), perennial herbaceous plant belonging to the family Hypericaceae. Organic and aqueous extracts of HP have been used in folk medicine and in pre-clinical testing for the treatment and prevention of several diseases through effects nefroprotetores, antioxidant, antifungal, anxiolytic, wound healing and antiviral activities. Clinical studies indicated that this specie can be useful in the treatment of central nervous system disorders, especially to unipolar depression. HP contains at least ten classes of biologically active compounds, including anthraquinones/naftodiantronas, phloroglucinol derivatives, flavonoids, biflavones, xanthones, volatile oils, amino acids, vitamin C, coumarins, carotenoids and tannins. At the same time that the secondary metabolites have important pharmacological effects, they can impair the effectiveness of other drugs by pharmacokinetic antagonism.

Hypericin: chemical synthesis and biosynthesis / 中国天然药物

Hypericin is one of the most important phenanthoperylene quinones extracted mainly from plants of the genus Hypericum belonging to the sections Euhypericum and Campylosporus of Keller's classification. Widespread attention to the antiviral and anti-tumor properties of hypericin has spurred investigations of the chemical synthesis and biosynthesis of this unique compound. However, the synthetic strategies are challenging for organic and biological chemists. In this review, specific significant advances in total synthesis, semi-synthesis, and biosynthesis in the past decades are summarized.

Determination of Hypericin in Rat Plasma by HPLC-MS/MS / 中国药师

Objective: To establish an HPLC-MS/MS method for the determination of hypericin in rat plasma. Methods: The sample was analyzed by HPLC-MS/MS after the addition of internal standard and then protein precipitation using acetonitrile. The sepa-ration was carried out on an Ultimate C18 column (150 mm × 2. 1 mm,5. 0 μm). The mobile phase was composed of acetonitrile∶ 5 mmol·L-1 ammoniumacetate (containing 0. 1% formic acid) (90∶10) at a flow rate of 0. 5 ml·min-1 under 35℃. The detection was performed with multiple teactions monitoring ( MRM) using an electrospray ionization ( ESI) . The precursor/product ion transitions were monitored at m/z 503. 2→m/z 405. 1 for hypericin and m/z 355. 0→m/z 41. 9 for the internal standard pioglitazone ( anion mode). Results:The good linearity of hypericin was obtained within the range of 0. 1-13. 2 ng·ml-1. The correlation coefficient was more than 0. 99 and the lower limit of quantification was 0. 1 ng·ml-1. The extraction recovery was within the range of 84. 19%-98. 71%. The precision of intra-and inter-day was below 18. 47%. Conclusion: The method is fast, sensitive and accurate, which provides research basis for the clinical further study of hypericin.

Content Determination of Hypericin in Guanye Jinsitao Drop Pills by HPLC / 中国中医药信息杂志

Objective To build a method for content determination of Hypericin in Guanye Jinsitao drop pills by HPLC.Methods The column was Kromasil C18 (150 mm×4.6 mm, 5μm), mobile phase was acetonitrile-0.1% phosphoric acid (16∶84), UV detection was performed at 360 nm, the flow rate was 1.0 mL/min, and the detector drift tube temperature was set at 30℃.Results The calibration curves were linear in the range of 0.132-1.188μg for Hypericin (r=0.999 9). The average recovery was 99.55% (RSD=0.78%).Conclusion The method is simple, accurate, reliable, and can be used for the quality control of Hypericin in Guanye Jinsitao drop pills.

Plantas con actividad fotosensibilizadora y potencial terapéutico en leishmaniasis cutánea: hipericina, una alternativa prometedora / Plants with photosensitizing activity and therapeutic potential in cutaneous leishmaniasis: Hypericin, a promising alternative

La leishmaniasis es una enfermedad endémica en 98 países, con más de 350 millones de personas en riesgo de adquirir la infección y 12 millones de personas infectadas. Numerosas desventajas asociadas a los tratamientos actuales promueven la falta de adherencia o abandono del tratamiento y la aparición de cepas resistentes al medicamento. Estos factores han estimulado la búsqueda de alternativas terapéuticas que sean rápidas, seguras, de fácil administración y económicas. Surge así la etnobotánica, que en combinación con la fototerapia puede convertirse en una estrategia innovadora para la búsqueda de compuestos de origen natural con actividad leishmanicida, donde productos como la hipericina pueden ser considerados candidatos prometedores en el descubrimiento y desarrollo de nuevos tratamientos. La hipericina es una molécula de origen natural que presenta un rendimiento cuántico alto en su estado triplete y genera eficientemente especies reactivas de oxígeno, lo cual posiblemente la hace un buen agente leishmanicida al ser aplicado en los tejidos infectados, además de ser posiblemente un potencial agente cicatrizante, que otorga un efecto cosmético favorable en la resolución de la lesión, con amplias ventajas como bajo costo y fácil manejo, convirtiéndose en una alternativa favorable frente a otras usadas en el manejo de la enfermedad.

Leishmaniasis is a disease endemic to 98 countries, with over 350 million people at risk of acquiring the infection and 12 million people already infected. The numerous disadvantages associated with current treatments encourage a lack of adherence and even abandonment of the disease treatment, resulting in the emergence of drug-resistant strains. These factors have stimulated the search for therapeutic alternatives that are fast, safe, easy to administer and economical. This has resulted in the emergence of ethnobotany, which, along with phototherapy, could become an innovative strategy for finding naturally occurring compounds with leishmanicidal activity. In this context, products such as hypericin could be considered promising candidates in the discovery and development of new treatments. Hypericin is a naturally occurring molecule that has a high quantum yield in its triplet state and efficiently generates reactive oxygen species. These properties could make hypericin an effective leishmanicidal agent when applied over infected tissues and a potential healing agent that provides cosmetic effects favorable to lesion resolution. In addition, its significant advantages in terms of low cost and easy handling make hypericin a favorable alternative to other treatments for managing this disease.

Effect of hypericin on myocardial fibrosis of chronic viral myocarditis of mice and its mechanism / 中草药

Objective: To investigate the role of JAK/STAT signal pathway in myocardial fibrosis (MF) of the chronic viral myocarditis (VMC) of mice and hypericin intervention study. Methods: Sixty healthy male Balb/c mice were used to establish the MF model by intermittent multiple ip injection of coxsackie B3, another 10 mice were used as normal control. Two months after the modeling, survival mice were randomly divided into four groups, model group, low- or high-dose hypericin group, and Captopril group. The mice were treated by Captopril or hypericin, respectively, ig administration, once a day. After 30 d, we took the myocardium of left ventricle to dye with Masson, to observe the cardiac histological changes. The serum type I collagen and type III collagen were detected by the means of ELISA, and the expression of JAK1 and STAT3 was observed with semi-quantitative RT-PCR and immunohistochemistry technique. Results: The model group, serum type I and type III collagen increased significantly, the expression level of JAK1/STAT3 was higher than that of the normal group, and the difference was significant (P < 0.05). While the hypericin and Captopril treatment could significantly reduce serum expression of type I and, type III collagen, and decrease JAK1/STAT3 expression. Histology showed the improvement of myocardial fibrosis degree, and a significant difference was observed when comparing with the model group (P < 0.05). Conclusion: In the process of chronic viral myocarditis, the activation of JAK1/STAT3 pathway may be one of pathological mechanisms of the MF-induced with type I and type III collagen increasing. Hypericin could inhibit the myocardial fibrosis by blocking JAK1/STAT3 pathway.

Application of hypericin in cancer therapy and diagnosis / 国际药学研究杂志

Hypericin, used as a medicinal heifo since ancient times, is a natural photosensitizer derived from the plant Hypericum perforantum. Hypericin would produce peroxides to induce apoptosis and inhibit the growth of oncocytes. Meanwhile, hypericin has a specific affinity with tumor pathology organization (underlying accumulation in pathology organization) which has been widely used in optical diagnosis. The present review gives a comprehensive summary of the anticancer effect based on photodynamic therapy, and photodynamic diagnosis of hypericin optical properties, with the purpose of making full use of the hypericin natural resources, and providing a basis for research and development of hypericin derivatives. © 2006 Editorial office of Foreign Medical Sciences.

The effects of hypericin on inhibition of hepatitis B virus replication in vitro / 中华传染病杂志

Objective To explore the effects of hypericin on inhibition of hepatitis 15 virus (HBV) replication.Methods The concentration gradient hypericin was added to HepG2.2.15 cell culture system and lamivudine was used as control.Enzyme-linked immunosorbent assay (ELISA) and Southern blot were used to examine HBsAg,HBeAg and HBV DNA level in the culture supernatant,respectively.Half inhibitory concentration (IC50) and half effective concentration (EC50) of hypericin were calculated.The effects of hypericin on HBV DNA polymerase were detected by 32p marked deoxy-ribonucleoside triphosphate as the substrate. Independent sample t-test and single factor analysis of variance were used to compare the data between two groups and among multiple groups.Results The inhibition rates of hypericin on HBsAg, HBeAg were enhanced with hypericin concentration increasing and those were higher than lamivudine control group when the concentration was higher than 0.5 μmol/L (t=-0.127,P＜0.05).Southern blot confirmed that hypericin was stronger in inhibition of HBV DNA (EC50=0.2 μmol/L) than lamivudine (t=-0.058,P＜0.05).Hypericin was non-toxic on HepG2.2.15 cells in the range of test with EC50 of 0.2 μmol/L and IC50 of 200 μmol/L.Hypericin did not act on HBV DNA polymerase which was quite different from lamivudine.Conclusions Hypericin can effectively inhibit HBV replication as well as antigen synthesis and is non-toxic on HepG2.2.15 cell.The anti-HBV target of hypericin is different from nucleos(t)ide analogues.

Long-term maintenance of in vitro cultures affects growth and secondary metabolism of St. John's Wort / A manutenção prolongada de culturas in vitro afeta o crescimento e o metabolismo secundário de hipérico

Hypericum perforatum is a traditional medicinal plant with wound healing and antidepressant properties. Efficiency of micropropagation is often related to the long term maintenance of tissues in culture, which may alter the secondary metabolism of plants. The objective of this study was to evaluate growth and secondary metabolism of in vitro shoots of H. perforatum on short and long term maintenance of cultures (30 and 100 days). The effect of BA and NAA supplementation was evaluated during 30 days of culture. Adventitious shoots were cultivated on MS medium supplemented with 4.4mM BA alone or in combination with 0.05mM NAA for 30 days. A hormone-free medium was used as control. Shoots cultivated for 100 days were maintained in presence of 4.4mM BA. Biomass, multiplication of shoots, contents of phenolic compounds, flavonoids and hypericin were evaluated. No difference between BA and BA+NAA was observed on growth, multiplication of shoots and levels of flavonoids at the end of 30 days of culture. Production of phenolic compounds was promoted by addition of BA+NAA to the medium, whereas hypericin was increased by the presence of BA. The time of culture (30 and 100 days) affected all the parameters analyzed, except the levels of flavonoids in the short term experiment.

Hypericum perforatum é uma planta medicinal que apresenta propriedades cicatrizante e antidepressiva. Frequentemente, a eficiência da micropropagação está relacionada à manutenção dos tecidos em cultura por longos períodos, o que pode alterar o metabolismo secundário das plantas. O objetivo deste trabalho foi avaliar o crescimento e o metabolismo secundário de brotações adventícias de H. perforatum mantidas por diferentes tempos de cultivo (30 e 100 dias). O efeito da adição de BA e ANA foi avaliado no período de 30 dias. As brotações foram cultivadas em meio MS suplementado com 4,4mM BA como único regulador ou em combinação com 0,05mM ANA, por 30 dias. Um meio de cultivo sem adição de reguladores foi utilizado como controle. As brotações cultivadas por 100 dias foram mantidas em presença de 4,4mM BA. A biomassa, a multiplicação dos brotos, as concentrações de compostos fenólicos, flavonoides e hipericina foram os parâmetros avaliados. Nenhuma diferença entre a adição de BA ou BA+ANA foi observada quanto ao crescimento, ao número de brotos e aos níveis de flavonoides ao final de 30 dias de cultivo. Diferenças neste período foram detectadas nos níveis de compostos fenólicos e de hipericina quando os brotos foram cultivados em presença de BA+NAA e de BA, respectivamente. O tempo de cultivo (30 e 100 dias) afetou todos os parâmetros avaliados, com exceção dos níveis de flavonoides no período de 30 dias.

Studies on the Secretory Structure of Hypericum erectum / 中草药

The structures that secrete and store hypericin and volatile oil, the main medicinal com-ponent of Hepericum erectum Thunb. , were studied using Sudan black, 5% NaOH solution and 0.5%methanolic magnesium acetate on semi-thin sections of H. erectum. Results showed that hypericin was pro-duced and stored in the secretory cell nodules, while the volatile oil was produced and stored in its secreto-ry sac and canal. The distribution and denseness of these 3 secretory structures in various organs were de-scribed to provide a scientific basis for the rational utilization of different parts of this medicinal plant.