Distinct Hypericum perforatum L. total extracts exert different antitumour activity on erythroleukemic K562 cells

Total flower extracts of Hypericum perforatum L. obtained with 3 different solvent systems were tested on tumour cell line cultures by comparing two groups of plants harvested in different times and places. The extracts, characterized according to the spectroscopic profile and the hypericin content, were tested on the growth and apoptotic death of K562 cells, a human erythroleukemic cell line. Growth and apoptosis were analysed by viable cell count, flow cytometry, and fluorescence microscopy at 6, 24, and 48 hr of culture following 1 hr exposure to the extracts under investigation. Here, we show that Hypericum extracts are able to reduce the growth of K562 cells and induce different degrees and kinetics of apoptosis according to the group of plants of origin. Also, we highlighted interesting differences in terms of efficacy among the extracts, with some samples losing their effectiveness along the culture time and others able to maintain or even increase their efficacy. Furthermore, the data herein obtained confirm the role of non hypericin compounds that are present in different proportions in the two plant groups and in the extracts analysed.     

Sedative activity in mice of a hydroalcohol extract of Hypericum perforatum L.

The sedative activity of a hydroalcohol extract of Hypericum perforatum L. (Hypericaceae) and some of its isolated constituents was evaluated in mice. A dry hydroalcohol extract, orally administered at 26.5 mg/kg, induced a marked sedation as measured by two different tests (actimeter and sleep potentiation), compared with controls (diazepam, 2 mg/kg, per os). Several fractions were isolated from this hydroalcohol extract: hypericin, flavones, butanol fraction, aqueous fraction; a separation and purification method was developed for hypericin. None of the isolated fractions exhibited the same sedative activity as the hydroalcohol extract. © 1997 John Wiley & Sons, Ltd.     

Hepatoprotective activity of Hypericum perforatum L. alcoholic extract in rodents

In recent years, interest in the hepatoprotective plants and plant based chemicals is increasing. In Turkish folk medicine, Hypericum perforatum L. has been used for various purposes including liver protection. The hepatoprotective effect of H. perforatum was investigated in vivo by cannulating the rat bile duct for choleretic activity and by barbiturate sleeping time following CCI₄-induced hepatic injury. The increase in the bile secretion following intraduodenal injection of H. perforatum extract, and the appearance of its constituents in the bile were observed by its colour change and this observation was confirmed by thin layer chromatographic analysis suggesting the excretion of hypericin into the bile as well. It was observed that the extract shortens the barbiturate sleeping time of CCI₄-treated mice suggesting hepatoprotection.     

贯叶连翘提取物抗抑郁作用研究

目的：研究贯叶连翘提取物对小鼠的抑制郁作用。方法：选取了强迫小鼠游泳实验，小鼠尾悬吊应激实验，拮抗利血平所致的抑制症状等实验指标。结果：贯叶连翘提取物150mg/kg,300mg/kg显著缩短迫游泳小鼠及尾悬吊小鼠的不动时间；显著拮抗利血平所致的体温下降作用和小鼠眼睑下垂作用。结论：贯叶连翘提取物具有一定的抗抑郁作用。     

大孔吸附树脂纯化贯叶连翘提取物的工艺

目的筛选纯化贯叶连翘提取物的最佳树脂型号及其最佳工艺。方法以金丝桃素为指标,对大孔吸附树脂的类型进行筛选,并采用正交试验,对最佳大孔吸附树脂工艺进行考察。结果最佳大孔吸附树脂为LSA-10,其最佳工艺为比上柱量1.954 2 mg/g;径高比1∶20;药液浓缩比1∶1,药液上样体积流量2 BV/h,分别以4 BV蒸馏水、3 BV 20%乙醇、8 BV 70%乙醇为洗脱剂洗脱,贯叶连翘提取物中的金丝桃素量提高到1.5%以上。结论该工艺合理、稳定,纯化效果好。     

贯叶金丝桃愈伤组织的诱导及其有效成分的定性

目的 :诱导出贯叶金丝桃的愈伤组织 ,并确定其中含有金丝桃素。方法 :在不同条件下采用组织培养技术 ,进行贯叶金丝桃的愈伤组织诱导研究 ,用HPLC方法确定其中含有的金丝桃素。结果和结论 :最佳诱导条件是MS基本培养基加入生长调节物质 2 ,4 D(4μg·L- 1 )和 6 BA(0 .2 μg·L- 1 )。叶腋等分生组织较多的部位易于诱导愈伤组织。诱导出的愈伤组织 ,经HPLC方法确定其含金丝桃素     

Antiinflammatory activity of ethanol extracts of Hypericum perforatum L., H. barbatum Jacq., H. hirsutum L., H. richeri Vill. and H. androsaemum L. in rats

Ethanol extracts of the aerial parts of six Hypericum species, H. barbatum, H. androsaemum, H. richerii, H. hirsutum, H. perforatum and H. perforatum cultivated on mountain Tara were tested for antiinflammatory activity in comparison with indomethacin (IND) using the carrageenan-induced rat paw edema test. It was found that all examined extracts produced antiinflammatory activity, particularly those from H. hirsutum, and both wild and cultivated H. perforatum. A dose-dependent antiinflammatory effect was especially pronounced in extracts showing some lower antiinflammatory activity. There was no correlation between the amount of hypericin in the extracts and their antiinflammatory activity. Copyright (c) 2006 John Wiley & Sons, Ltd.     

贯叶金丝桃不同器官的分泌细胞团分布密度与金丝桃素含量的相关性研究

目的 研究贯叶金丝桃不同器官的分泌细胞团分布密度和金丝桃素含量及其相关性，为选择药用成分高的品种和合理的采收部位提供依据。方法 在显微镜下观察透明装片，统计叶、花萼和花瓣中分泌细胞团的分布密度，用HPLC法分析各器官中金丝桃素含量。结果 根、茎、果实和叶的中部不含金丝桃素类物质，叶缘部分含金丝桃素0.1456％，花萼含0.0653％，花瓣含1．2682％。结论 具有分泌细胞团的器官或部位含有金丝桃素类物质，且与其分布密度成正相关；而不具分泌细胞团的器官或部位，则不含有金丝桃素类物质。     

The inhibitory effect of Hypericum perforatum extract on morphine withdrawal syndrome in rat and comparison with clonidine

The effects of Hypericum perforatum (St John's wort) on morphine withdrawal syndrome and comparison with clonidine have been investigated in morphine‐dependent rats. Adult male Wistar rats were subjects. The frequencies of withdrawal behavioral signs (rearing, teeth chattering and jumping) induced by naloxone challenge were demonstrated in morphine dependent rats receiving Hypericum perforatum extract (HPE), saline or clonidine. The withdrawal behavioral manifestations in rats were inhibited significantly by chronic co‐administration of Hypericum perforatum extract or clonidine with morphine. This study showed that clonidine was more effective than HPE at a dose of 0.4 mL/200 g and there was no significant statistical difference between the mean frequency of withdrawal signs of HPE at a dose of 0.8 mL/200 g compared with clonidine (0.2 mg/kg i.p.) but at a dose of 1.2 mL/200 g of HPE was significantly stronger than clonidine in attenuation of the morphine withdrawal syndrome. The findings suggest that HPE is capable of reducing the symptoms of opiate withdrawal and its effectiveness may be equivalent to clonidine in reducing the opiate withdrawal syndrome and may have human therapeutic potential. Copyright © 2009 John Wiley & Sons, Ltd.     

贯叶连翘组织培养及植株再生研究

在国内首次研究了贯叶连翘胚轴和子叶的培养方法,分别通过器官型和器官发生型途径获得了大量再生植株,部分可移栽成活,实验表明,在ＢＡ和２,４－Ｄ不同组合的ＭＳ培养基上,愈伤组织易于诱导,繁殖速度快,并且通过初步检测证明了愈伤组织中金丝桃素存在的可能性,再生植株产生极容易,繁殖系数高。     

贯叶连翘中金丝桃素的提取及含量测定

目的:优选贯叶连翘中金丝桃素的提取工艺,并建立其含量测定方法。方法:考察高效液相色谱法测定金丝桃素含量的条件;以金丝桃素含量为指标,考察提取溶剂、提取时间、液料比、溶剂浓度等因素对超声提取金丝桃素的影响,筛选出最佳工艺。结果:确定最佳工艺为液料比:150∶1、溶剂:100%甲醇、提取时间30min;金丝桃素在0.0104～0.208μg(r2=0.9999)线性关系良好,平均回收率98.8%,RSD=0.54%。HPLC的色谱条件为:色谱柱:依利特HypersilODS2(5.0mm×200mm,5μ);流动相:甲醇-0.1mol.L-1 NaH2PO4(97∶3);柱温:30℃;检测器:SPD-20A UV/VIS;检测波长:590nm。结论本研究建立的方法稳定、准确、可行,可用于今后金丝桃素的研究。     

贯叶连翘提取物中金丝桃苷的分析鉴别与稳定性研究

目的探讨贯叶连翘提取物中金丝桃苷鉴别方法和稳定性。方法对贯叶连翘提取物进行化学、薄层、紫外鉴别。通过加速实验从不同影响因素分析贯叶连翘提取物中金丝桃苷的稳定性。结果贯叶连翘提取物的盐酸-镁粉反应显阳性,薄层色谱斑点清晰,紫外光谱在363 nm处有最大吸收峰。贯叶连翘提取物在酸、碱、热、湿、光、氧化、加速实验等条件下金丝桃苷的含量均有不同程度的损失。结论3种鉴别方法能很好地鉴别出黄酮苷类(金丝桃苷)的存在,定性方法简便、准确、专属性强。鉴于金丝桃苷的稳定性,在对贯叶连翘进行贮存时,应密闭、闭光、置阴凉干燥处保存。     

Antiviral In vitro activity of Hypericum perforatum l. extract on the human cytomegalovirus (HCMV)

The total methanol extract and chromatographic fractions of the extract of the aerial parts of Hypericum perforatum L. were investigated for their antiviral activity against human cytomegalovirus (HCMV) AD-169 laboratory strain by modification of the widely used diagnostic method ‘shell-vial’ culture. The plant selection was based on its traditional use against viruses of the Herpes family in Greece. The results showed extract fraction III had the most potent antiviral activity against HCMV which was also superior to that of ganciclovir. Copyright © 1998 John Wiley & Sons, Ltd.     

Wound-healing activity of St. John's Wort (Hypericum perforatum L.) on chicken embryonic fibroblasts

Wound-healing effect of St. John's Wort (Hypericum perforatum L.) extract was evaluated by comparing with dexpanthenol and titrated extract of Centella asiatica (TECA) on cultured chicken embryonic fibroblasts. Chicken embryonic fibroblasts from fertilized eggs were incubated with the plant extract, dexpanthenol and TECA. Using microscopical methods by staining cells, mitotic ability, morphologic changes and collagen production in the cultured fibroblasts were evaluated as parameters to approach its mechanism of action in wound repair. Findings obtained in the present study indicated that Hypericum perforatum extract exhibited a wound-healing activity whose mechanism of action is similar to that of TECA. Wound-healing activity of Hypericum perforatum extract seems to be mainly due to the increase in the stimulation of fibroblast collagen production and the activation of fibroblast cells in polygonal shape, which plays a role in wound repair by closing damaged area. The findings demonstrated the wound-healing activity of Hypericum perforatum, which has previously been based on ethnomedical data.     

正交试验优选贯叶连翘中金丝桃素的提取方法及因素

目的:对贯叶连翘中金丝桃素的提取方法及相关影响因素进行探讨.方法:以金丝桃素含量为指标,分别考察冷浸法、超声波提取法、回流提取法对提取率的影响,以及液料比、超声时间对提取率的影响,并对提取方法(A)、液料比(B)和提取时间(C)3因素设计正交试验.结果:影响金丝桃素提取率的主次因素为A>B>C,即提取方法>液料比>提取...     

Cytotoxic effects of the Viscum album L. extract on Ehrlich tumour cells in vivo

To date most pharmacological studies on mistletoe (Viscum album L.) have focused on the therapeutic properties of its polar extracts. This study examined the non-polar constituents of Viscum album and their biological activities. Supercritical CO(2) extraction coupled with gas chromatography/mass spectrometry (GC/MS) was used to selectively extract and identify compounds in Viscum album leaves. Several non-polar classes of compounds were identified in the extract. In addition, a volatile fraction was identified that contained several novel terpene molecules. The hypothesis was tested that the Viscum album extract exhibits cytotoxic properties in Ehrlich carcinoma (EAC) cells in vivo due to the induction of oxidative stress. A significant reduction in the incidence of cancer was observed in all groups that received the Viscum album extract compared with the EAC control group. The largest decrease was observed in mice pretreated with the Viscum album extract, although significantly reduced numbers of EAC cells were also observed in animals with developed carcinoma. The activities of antioxidative enzymes in the EAC cells suggested the absence of oxidative stress. However, changes in the antioxidative enzymes activities observed after administration of the Viscum album extract might be due to the induction of oxidative stress in the EAC cells.     

Antiinflammatory and antioxidant activity of Hypericum rumeliacum Boiss. subsp. apollinis (Boiss. & Heldr.) Robson & Strid methanol extract

Plants of the genus Hypericum are widely used in folk medicine for the treatment of gastric ailments (including pectic ulcers), burns, swelling, inflammation, anxiety, as well as for bacterial and viral infections. The aerial parts of Hypericum rumeliacum Boiss. subsp. apollinis (Boiss. & Heldr.) Robson & Strid (Guttiferae), a perennial herb growing wild in Greece, were subjected to biological and phytochemical studies. The H. rumeliacum methanol extract showed, on the DPPH test, antioxidant activity (IC(50) = 23.61 microg/mL). The antiinflammatory activity of the extract (70 mg/kg, i.p.), in the carrageenan-induced paw oedema in rat, was studied. In this experimental model an inhibition of paw oedema, significant from the first to third hour, was observed. The histological examination confirmed resolution of the inflammatory process observed in vivo. Through HPLC analysis of the H. rumeliacum methanol extract, the naphthodianthrones hypericin, pseudohypericin and some polyphenol compounds, such as chlorogenic acid, rutin and isoquercitrin, were identified and quantified. The antioxidant activity of H. rumeliacum could be chiefly related to polyphenol compounds that are well known as the main contributors to the free radical-scavenging effects. The antiinflammatory effect of the methanol extract of the aerial parts could be due to the synergic action both of hypericin and polyphenol compounds.     

Effects of Hypericum perforatum extract on diabetes‐induced learning and memory impairment in rats

Cognitive impairment occurs in diabetes mellitus. Hypericum perforatum has been used in folk medicine to improve mental performance. Here it is hypothesized that chronic treatment with an extract of Hypericum perforatum (6, 12 and 25 mg/kg, p.o.) would have effects on passive avoidance learning (PAL) and memory in control and streptozotocin‐induced diabetic rats. Treatments were begun at the onset of hyperglycaemia. PAL was assessed 30 days later. A retention test was done 24 h after training. At the end, the animals were weighed and blood samples were drawn for plasma glucose measurement. Diabetes caused impairment in acquisition and retrieval processes of PAL and memory. Hypericum treatment (12 and 25 mg/kg) improved learning and memory in control rats and reversed learning and memory deficits in diabetic rats. A dose of 6 mg/kg did not affect cognitive function. Hypericum administration did not alter the body weight and plasma glucose levels. Antioxidant properties and cholinergic facilitatory effects of Hypericum may be involved in its nootropic effects. These results show that Hypericum perforatum prevented the deleterious effects of diabetes on PAL and memory. As Hypericum would be free of major side effects compared with other nootropic medications, it may provide a new potential alternative for demented diabetic patients. Copyright © 2010 John Wiley & Sons, Ltd.     

Effects of St John's wort (Hypericum perforatum L.) extract on plasma androgen concentrations in healthy men and women: a pilot study

St John's wort extract (SJW; Hypericum perforatum L.) is taken extensively as a putative herbal antidepressant. It has been shown to induce the activity of cytochrome P-450 3A4 (CYP3A4) and to increase the clearance of numerous drugs and steroids such as cortisol and ethinyl estradiol. This study was conducted to determine if SJW exposure also alters the concentrations of circulating androgenic steroid hormones. The study was conducted using healthy volunteers (6M, 6F) studied before and after a 14-day treatment period with a SJW preparation previously demonstrated to induce the activity of CYP3A4. Plasma concentrations of testosterone, dihydrotestosterone (DHT), dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG) and the combined concentrations of androsterone sulfate (AoS) and epiandrosterone sulfate (epiAoS) were measured by immunoassay methods. The results of analysis demonstrated that SJW did not significantly alter the majority of the androgens studied (p > 0.05) although the combined concentrations of the 5alpha-reduced steroids, AoS and epiAoS, significantly declined following treatment in all subjects (p = 0.02), and in males (p = 0.04). Furthermore, the testosterone to DHT ratio was increased in both men and women. Although the latter increase did not reach statistical significance, it is also consistent with the possible inhibition of 5alpha-reductase by SJW. It is concluded that despite significant induction of CYP3A4, short term administration of SJW does not significantly alter the concentrations of most circulating androgens in men and women but may produce a dimunition in some of the circulating 5alpha-reduced androgens.     

Evaluation of in-vivo wound healing activity of Hypericum patulum (Family: hypericaceae) leaf extract on different wound model in rats

The methanol extract of Hypericum patulum Thumb. leaves were investigated for the evaluation of their wound healing potential on different experimental models of wounds in rats. The methanol extract of leaves (HPM), in the form of an ointment with two different concentrations (5% and 10% w/w ointment of leaf extract in simple ointment base) was evaluated for wound healing potential in an excision wound model and an incision wound model in rats. Both concentrations of the methanol extract ointment showed significant responses in both the wound types tested when compared with the control group. The effect produced by the extract ointment, in terms of wound contracting ability, wound closure time, regeneration of tissues at wound site, tensile strength of the wound and histopathological characteristics were comparable to those of a standard drug nitrofurazone ointment.