贯叶金丝桃(Hypericum perforatum)种子灌浆与发芽特性研究

贯叶金丝桃(Hypericum perforatum)是抗抑郁中药的基原植物,以有性繁殖为主,种子微小。为了揭示贯叶金丝桃种子发育规律,明确种子适宜采收期,以甘肃礼县栽培的2年生贯叶金丝桃为研究对象,于开花后第5天开始,每隔3 d测定种子粒重变化动态,并拟合Logistic曲线方程,最后测定各时期采收种子的发芽指标。结果表明:贯叶金丝桃种子灌浆持续期44 d,籽粒干质量变化符合Logistic生长曲线:开花后第5~11天为渐增期,第11~32天为快增期,第32~44天进入稳增期后略有下降。在灌浆过程中,种子鲜质量呈先升高后下降的动态变化趋势,在花后第32天达到最大值0.2697 g;灌浆速率和脱水速率呈波动趋势,种子干质量与平均灌浆速率、灌浆持续期呈极显著正相关,与含水量呈极显著负相关。种子发芽率和发芽指数在开花后44天达到最大,发芽势在开花后41天达到最大,发芽率、发芽势和发芽指数均与灌浆持续期呈极显著正相关,与含水量呈极显著负相关。以上说明甘肃礼县贯叶金丝桃繁种时以开花后44天左右(8月9日前后)为种子采收适宜期,过早采收和延迟采收均影响种子发芽质量。

In Vitro and in Silico Insights on the Biological Activities, Phenolic Compounds Composition of Hypericum perforatum L. Hairy Root Cultures

Three Hypericum perforatum hairy root lines(HR B,HR F and HR H)along with non-transformed roots were analyzed for phenolic compounds composition and in vitro enzyme inhibitory properties.In silico molecular modeling was performed to predict the interactions of the most representative phenolic compounds in HR clones with enzymes related to depression,neurodegeneration and diabetes.Chromatographic analyses revealed that HR clones represent an efficient source of quinic acid and hydroxybenzoic acids,epicatechin and procyanidin derivatives,quercetin and kaempferol glycosides,as well numerous xanthones.In vitro antidepressant activity of HR extracts through monoamine oxidase A(MAO-A)inhibition was attributed to the production of oxygenated and prenylated xanthones.The neuroprotective potential of HR extracts was related to the accumulation of quercetin 6-C-glucoside,epicatechin,procyanidins andγ-mangostin isomers as potential inhibitors of acetylcholinesterase(AChE)and butyrylcholinesterase(BChE).Vanillic acid and prenylated xanthones in HR clones as promising inhibitors of tyrosinase additionally contributed to the neuroprotective activity.Five preeminent xanthones in HR(γ-mangostin,mangiferin,garcinone C,garcinone E and 1,3,7-trihydroxy-6-metoxy-8-prenyl xanthone)along with the flavonol quercetin 6-C-glucoside effectively inhibitedα-amylase andα-glucosidase indicating the antidiabetic properties of HR extracts.Transgenic roots of H.perforatum can be exploited for the preparation of novel phytoproducts with multi-biological activities.

A New Antifungal Flavonol Glycoside from Hypericum perforatum

In addition to six known flavonoids quercitrin, hyperoside, avicularin, rutin, quercetin and kaemferol, a new flavonol glycoside named 6″_O_acetyl quercetin 3_O_β_ D _alloside (1) was isolated from the aerial parts of Hypericum perforatum L. The structures were determined on the basis of spectroscopic methods (UV, IR, FAB_MS, 1H_NMR and 13 C NMR). Antifungal assay of all compounds showed that metabolite 1, quercitrin and quercetin were inhibitory to the growth of phytopathogenic fungus Helminthosporium sativum Pamel King et Bakke with minimum inhibitory concentrations (MICs) of 25, 50 and 50 μg/mL, respectively. Moreover, glycoside 1 and quercitrin were also shown to be able to inhibit the growth of Fusarium graminearum Schw. with MIC of 100 μg/mL. The MICs of ketoconazole used as control against the test fungi were 0.5 μg/mL in our assay.

Ultrastructure of the Multicellular Nodules in Hypericum perforatum Leaves

With the development of the multicellular nodules in the leaves of Hypericum per plastids increased both in number and volume, and some plastids degenerated and were surrounded by dark tubular elements and vesicles. Some vesicles fused with vacuoles and secreted substances into vacuoles. There are many multivesicles, multimembrane structures and osmiophilic droplets in vacuoles of nodule cells. Meanwhile, dictyosome secreted vesicles into the vacuoles. However, there is a large central vacuole completely filled with secretory materials (hypericin) in matured multicellular nodules. This suggests that the osmiophilic droplets may be a precursor of hypericin originated from the degenerated plastids. There were abundant endoplasmic reticulums and dictyosomes between the plastids and osmiophilic droplets, suggesting that they may be involved in the synthesis and/or transport of the precursor of hypericin.

An Isopropyldioxy Flavonol from Hypericum perforatum L.

Aim Isolation and structural elucidation of the constituents from the aerialpart of Hypericum perforatum L. Methods Column chromatography and TLC were used to isolate chemicalconstituents. Physico-chemical characterization and spectroscopic analysis were employed forstructural identification. Results Three flavonols were isolated and identified. Conclusion Compound1 is a novel natural product and its structure has been characterized to be 3, 5, 7-trihydroxy-3',4'-isopropyldioxy-flavone.

Antihyperglycemic effect of Hypericum perforatum ethyl acetate extract on streptozotocin-induced diabetic rats

Objective:To evaluate the antihyperglycemic activity of ethyl acetate extract of Hypericum perforatum(H.perforatum)in streptozotocin(STZ)-induced diabetic rats.Methods:Acute toxicity and oral glucose tolerance lest were performed in normal rats.Male albino rats were rendered diabetic by ST/(40 mg/kg,intraperitoneally).H.perforatum ethyl acetate extract was orally administered to diabetic rats at SO,100 and 200 mg/kg doses for 15 days to determine the antihyperglycemic activity.Biochemical parameters were determined at the end of the treatment.Results:H.perforatum ethyl acetate extract showed dose dependant fall in fasting blood glucose(FBG).After 30 min of extract administration,FBG was reduced significantly when compared with normal rats.H.perforatum ethyl acetate extract produced significant reduction in plasma glucose level,serum total cholesterol,triglycerides,glucose-6-phosphatase levels.Tissue glycogen content,HDL-cholesterol,glucose-6-phosphate dehydrogenase were significantly increased compared with diabetic control.No death or lethal effect was observed in the toxic study.Conclusions:The results demonstrate that H.perforatum ethyl acetate extract possesses potent antihyperglycemic activity in STZ induced diabetic rats.

Influence of Hypericum perforatum Extract on Piglet Infected with Porcine Respiratory and Reproductive Syndrome Virus

To study the influence of Hypericum perforatum extract （HPE） on piglets infected with porcine respiratory and reproductive syndrome virus （PRRSV）, enzyme-labeled immunosorbent assay （ELISA） and cytopathic effect （CPE） were used to determine in vitro whether HPE could induce swine pulmonary alveolar macrophages （PAMs） to secrete IFN-γ, and whether PRRSV titers in PAMs were affected by the levels of HPE-induced IFN-γ. HPE （200 mg·kg-1） was administrated by oral gavage to piglets infected with the PRRSV in vivo to observe whether HPE affected the viremia, lung viral titers, and weight gain of piglets infected with PRRSV. The results showed that HPE was capable of inducing PAMs to produce IFN-γ in a dose dependent manner and HPE pretreatment was capable of significantly reducing PRRSV viral titers in PAMs （P〈 0.01）. Administration of HPE to the PRRSV-infected animals significantly （P〈 0.05） reduced viremia over time as compared with the PRRSV-infected animals. But there was not significant decrease in lung viral titers at day 21 post-infection between the HPE- treated animals and the PRRSV-infected control piglets. There were no significant differences in weight gain over time among the HPE-treatment animals, the normal control, and the HPE control animals. The PRRSV-infected animals caused significant （P〈 0.01） growth retardation as compared with the HPE controls and the normal piglets. It suggested that HPE might be an effective novel therapeutic approach to diminish the PRRSV-induced disease in swine.

Pyrohyperforin, A New Prenylated Phloroglucinol from Hypericum perforatum

Pyrohyperforin, a new prenylated phloroglucinol was isolated from Hypericum perforatum, its structure was established on the basis of spectral data.

In vitro Anti-Hepatitis B Virus Effect of Hypericum perforatum L.

The anti-hepatitis B virus(HBV)effects and its mechanisms of the ethanol extracts of Hypericum perforatum L.(EHP)in vitro were explored.HepG2 2.2.15 cells,a stable HBV-producing cell line,were cultured as the model system to observe the anti-HBV effect.The viral antigens of cellular secretion,HBsAg and HBeAg,were determined by enzyme linked immunosorbent assay(ELISA).The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR.In order to understand the mechanisms of the suppression of H...

Anti-Influenza A Virus Effect of Hypericum perforatum L. Extract

To study the antiviral effect of Hypericum perforatum L. extract （HPE） on influenza A virus （IAV） （H1N1） in vitro and in vivo. Cytopathic effect （CPE） and neutral red （NR） dye uptake were used to examine the antiviral effect of HPE on Madin Darby Canine Kidney （MDCK） cells which were infected with IAV in vitro HPE was effective against influenza A virus （IAV） in vitro, with a 50% effective concentration （EC50） of 40 ug/mL, The mean 50% cytotoxic concentration （CC50） in the MDCK used in these experiments was 1.5 mg/mL. Ribavirin was run in parallel with EC50 values of 5.0 ug/mL; the mean CC50 for ribavirin was 520 ug/mL. Oral gavage administrations of HPE or ribavirin to mice infected with the IAV were highly effective in preventing death, slowing the decline of arterial oxygen saturation, inhibiting lung consolidation and reducing lung virus titers. The minimum effective dose of HPE in these studies was 31.25 mg/kg/day, which was administered twice daily for 5 d beginning 4 h prior to virus exposure. Below a dosage of 2000 mg/kg/day, almost all treated mice survived, which suggests that HPE is of low toxicity. Ribavirin＇s minimum effective dose was 40 mg/kg/day with the LDso determined to be 200 mg/kg/day. Delay of the initiation of either HPE or ribavirin therapy, using approximately 1/3 LD50 dose each time, could still be protective as late as 48 h after exposure to the IAV. While both agents appeared to have similar efficacy against IAV infections, HPE was considered to be less toxic and may warrant further evaluation as a possible therapy for influenza.

Effect of the Extract from Hypericum perforatum on Highly Pathogenic Avian Influenza Virus(HPAIV)

[Objective] The aim was to study the effects of the extract from Hypericum perforatum on highly pathogenic Avian Influenza Virus(HPAIV)in vivo.[Method] Chickens infected with H5N1 virus were treated with the extract from H.perforatum for 4 d.The virus was then detected by hemoagglutination(HA)test and reverse transcription polymerase chain reaction(RT-PCR).[Result] No H5N1 virus could be detected at the 7th d when the chickens were treated with 0.2 or 0.1 g/(kg·d)of the extract from H.perforatum.However,the virus could be detected in other chickens without the extract from HPE treatment.[Conclusion] The extract from H.perforatum might be a potential drug candidate to control infection of H5N1 subtype AIV in chickens.

Recent enhancement of the immunity in AIDS and other immunocompromised patients by hyperforin an antibiotic from <i>Hypericum perforatum</i>L. (<i>in vitro</i>model) part I

Today, Hypericum perforatum L. is probably one of the best-characterized medicinal plants, and hyperforin is its best-characterized constituent. Extracts from H. perforatum are widely used as antidepressants;however, less attention has been given to other properties of hyperforin, such as antitumor, fungicidal, antiviral and antibacterial action, or its possible use as a substance with immunomodulation properties. The present study summarizes results that describe the influence of hyperforin as an immunomodulation agent on phagocytosis and the breakdown of Escherichia coli by human polymorphonuclear neutrophils (PMNs). Hyperforin at 1 - 100 μg/mL concentrations was found to have a major influence on phagocytosis and the breakdown of E. coli by PMNs in vitro. A 100 μg/mL solution of hyperforin increased the uptake of non-opsonized E. coli almost 50-fold, and the uptake of IgG-opsonized E. coli more than threefold;on the other hand, the uptake of serum-opsonized bacteria was reduced to approximately 60% of that of the control. Hyperforin seems to bind to both PMNs and E. coli and acts like an opsonin. The elimination of remnants of IgG-opsonized E. coli from the PMNs was stimulated by hyperforin, while the elimination of remnants from non-op-so nized and serum-opsonized material was unaffected by the drug. Hyperforin exhibited clear immunomodulation ability as a phagocytosisstimulating agent. Hyperforin is probably inactive against human immunodeficiency virus (HIV) and most Gram-negative bacteria. However, it can protect acquired immunodeficiency syndrome (AIDS) patients and other immunocompromised patients by its antibacterial activity against Gram-positive bacteria and by enhancement of phagocytosis of Gram-positive and Gram-negative bacteria;some Gram-negative bacteria, such as Neisseria, are sensitive to hyperforin. Hyperforin has the ability to penetrate the blood-brain barrier (BBB) and blood-testis barrier (BTB) and is a valuable antibacterial agent against meningitis and gonorrhea. These properties of hyperforin are important for an antibiotic with immunomodulation activity in the struggle against the growing mortality in AIDS patients as a result of opportunistic bacteria, as recently shown by Bekondi et al. (2006, Int. J. Infect. Dis. 10, 387-395). It could also help to combat primary and opportunistic pathogens associated with meningitis in adults' relation to HIV serostatus.

An extract of Hypericum perforatum induces wound healing through inhibitions of Ca^(2+) mobilizations,mitochondrial oxidative stress and cell death in epithelial cells:Involvement of TRPM2 channels

The wound is induced by several mechanical and metabolic factors.In the etiology of the wound recovery,excessive oxidative stress,calcium ion(Ca^(2+))influx,and apoptosis have important roles.Ca^(2+)-permeable TRPM2 channel is activated by oxidative stress.Protective roles of Hypericum perforatum extract(HP)on the mechanical nerve injury-induced apoptosis and oxidative toxicity through regulation of TRPM2 in the experimental animals were recently reported.The potential protective roles in HP treatment were evaluated on the TRPM2-mediated cellular oxidative toxicity in the renal epithelium(MPK)cells.The cells were divided into three groups as control,wound,and wound+HP treatment(75μM for 72 h).Wound diameters were more importantly decreased in the wound+HP group than in the wound group.In addition,the results of laser confocal microscopy analyses indicated protective roles of HP and TRPM2 antagonists(N-(p-Amylcinnamoyl)anthranilic acid and 2-aminoethyl diphenylborinate)against the wound-induced increase of Ca^(2+) influx and mitochondrial ROS production.The wound-induced increase of early(annexin V-FITC)apoptosis and late(propidium iodide)apoptosis were also decreased in the cells by the HP treatment.In conclusion,HP treatment acted protective effects against wound-mediated oxidative cell toxicity and apoptosis through TRPM2 inhibition.These effects may be attributed to their potent antioxidant effect.

Pleomorphic hepatocellular carcinoma following consumption of hypericum perforatum in alcoholic cirrhosis

Hepatocellular carcinoma (HCC) often develops in patients with underlying liver disease, yet HCC with syncytial giant cells (SGCs) is extremely rare. Herein, we report a 55-year-old man with a 6-year history of alcoholic cirrhosis who during his regular checkup presented with marked elevation of alpha-fetoprotein. Clinical examination and imaging analyses revealed a tumor-like lesion in segment 4 of the liver, which was removed by limited wedge resection. Histological analysis by hematoxylin and eosin staining indicated pleomorphic and atypical nodules, with some SGCs, embedded within the boundaries of the neoplastic lesion. The adjacent liver parenchyma showed microvesicular steatosis, pericellular fibrosis, and moderate hemosiderin accumulation (grade 2, as determined by Prussian blue iron stain) in hepatocytes and Kupffer cells but no copper accumulation (as determined by orcein stain). Immunohistochemical analysis showed hepatocyte antigen-positive staining for the neoplastic cells and SGCs. The diagnosis was made for cirrhosis-related HCC with SGCs. The previous reports of pleomorphic HCC have featured osteoclast-like (i.e., mesenchymal type) giant cells, making this case of epithelial type giant cells very rare. The patient&#x02019;s 6-month history of hypericum perforatum/St John&#x02019;s wort self-medication may have prompted the cirrhosis or HCC progression or the unusual SGC manifestation.

长柱金丝桃(Hypericum ascyron L.)分泌结构的研究

对长柱金丝桃各类器官进行了整体透明和解剖学研究,确定合理的采收部位和采摘时间。结果表明,植株富含分泌囊和分泌小管道两种分泌结构;叶片中分泌囊(道)的发育属于裂生型;地上部分泌结构分布较多,尤以叶片中数量最多、分布密度最大;确定植物的最佳采收时期是每年8～9月的花果期、最佳采收部位是地上部。此研究结果为此药用植物合理的采收部位和采收时间的确定提供解剖学依据,同时为其他植物分泌囊(道)发育规律的研究提供理论依据。

贯叶金丝桃化学成分的分离与鉴定

目的研究藤黄科(Guttiferae)金丝桃属(Hypericum)贯叶金丝桃(Hypericum perforatum L.)的化学成分。方法采用正相硅胶色谱、反相ODS色谱、凝胶色谱及高效液相色谱等多种色谱方法对贯叶金丝桃体积分数95%乙醇提取物进行分离纯化。利用紫外、红外、核磁共振光谱,高分辨质谱及X-ray单晶衍射法对化合物的结构进行鉴定。结果从贯叶金丝桃中分离鉴定8个化合物,分别为perforxanthone A(1)、8-hydroxy-1,2,3-trimethoxyxanthone(2)、绵马酚D(3)、(S)-5-hydroxy-3,4-dimethyl-5-pentylfuran-2(5H)-one(4)、ent-4(15)-eudesmene-1β,6α-diol(5)、methyl-(E)-8-oxooctadec-9-enoate(6)、模绕酮酸(7)、白桦脂酸(8)。结论化合物1为新的氧杂蒽酮类化合物,化合物2-7为首次从该植物中分离得到。

贯叶金丝桃二氯甲烷部位的化学成分研究

目的研究贯叶金丝桃Hypericum perforatum L.二氯甲烷部位的化学成分。方法贯叶金丝桃二氯甲烷部位采用硅胶、ODS、Sephadex LH-20、半制备HPLC等进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。结果从中分离得到11个化合物,分别鉴定为贯叶金丝桃降碳聚酮A(1)、α-芒柄花醇(2)、(3 R)-thunberginol(3)、2-geranyloxy-1-(2-methylpropanoyl)-phloroglucinol(4)、4,6-dihydroxy-2-O-(3″,7″-dimethyl-2″,6″-octadienyl)-1-(2′-methylbutanoyl)benzene(5)、norhyperpalum G(6)、garsubellin A(7)、garsubellin B(8)、(2″R/S)-kellerine C(9)、kobusone(10)、圣草酚(11)。结论化合物1为新化合物,化合物2~3为首次在藤黄科植物中分离得到,化合物4~10为首次从该植物中分离得到。

RP-HPLC法测定贯叶连翘片剂中有效成分金丝桃素和金丝桃苷的含量

目的 :采用RP -HPLC法测定贯叶连翘片剂中有效成分金丝桃素 (hypericin)和金丝桃苷 (hyperoside)的含量。方法 :分别使用ODS和ODS - 2色谱柱 ,流动相分别是 1 5 6%磷酸二氢钠 (用磷酸调 pH至 2 1) -甲醇 -醋酸乙酯 ( 1∶4∶1)及甲醇 - 0 5 %磷酸 ( 4 5∶5 5 ,用三乙胺调 pH至 3 0 ) ,检测波长分别为 5 90nm ,3 60nm。结果 :加样回收率分别为 98 3 2 % (n =5 ) ,96 0 4 % (n =5 )。金丝桃素的进样量为 0 0 0 98～ 0 16μg、金丝桃苷的进样量为 0 4 3～ 3 4 2 μg时 ,线性关系良好。结论

贯叶金丝桃药材中萘骈二蒽酮类成分的分光光度法和HPLC法定量分析

目的:建立测定贯叶金丝桃药材中萘骈二蒽酮类成分的紫外-可见分光光度法和HPLC法,比较两者测定结果的差异与定量可靠性。方法:紫外-可见分光光度法以金丝桃素为指标性成分,在590nm处测定。HPLC法以DiamonsilC18柱(150mm×4.6mm,5μm为色谱柱,甲醇-10mmol/L CH3COONH4水溶液(pH值为6.2)(90∶10)为流动相,流速1.0ml/min,检测波长590nm,柱温30℃。经紫外光谱与质谱确认其中主要色谱峰成分为伪金丝桃素和金丝桃素,并以伪金丝桃素、金丝桃素为对照品进行定量,萘骈二蒽酮大类成分含量以伪金丝桃素、金丝桃素含量之和来表征。结果:紫外-可见分光光度法定量金丝桃素在2.44～19.52μg/ml范围内线性关系良好(r=0.999 6,n=3),测得贯叶金丝桃药材中萘骈二蒽酮类成分以金丝桃素计为(0.990 2±0.006 0)μg/mg。HPLC法定量伪金丝桃素在0.402～8.032μg/ml范围内线性关系良好(r=0.999 5,n=3)),定量金丝桃素在0.488～9.760μg/ml范围内线性关系良好(r=0.999 7,n=3)),测得贯叶金丝桃药材中伪金丝桃素和金丝桃素含量分别为(0.387 2±0.001 4)、(0.220 2±0.000 7)μg/mg,两者的总量为(0.605 5±0.001 2)μg/mg。结论:对贯叶金丝桃药材中萘骈二蒽酮类成分的定量分析,紫外-可见分光光度法和HPLC法测定结果差异明显;即使化学结构与光谱特性相似的大类成分,其紫外-可见分光光度法的定量准确性仍需要验证与评价。

TLC-分光光度法测贯叶连翘中金丝桃素含量

:针对贯叶连翘中成分复杂 ,干扰物多的特点 ,采用薄层层析 (TL C)分离出金丝桃素后 ,再进行分光光度测定。薄层层析以石油醚 -乙酸乙酯 -甲醇 (1∶ 4∶ 3)为展开体系展开 ,金丝桃素的 Rf=0 .85。分光光度法测定的回归方程为 A=72 .0 0 C+0 .0 0 3(r=0 .9997) ,平均回收率为 97.9% (RSD=0 .87% )。