绞股蓝抗肿瘤的药理和临床研究进展

绞股蓝抗肿瘤的药理和临床研究进展潘扬,魏菁（南京中医药大学,南京２１００２９）绞股蓝Ｇｙｎｏｓｔｅｍｍａｐｅｎｔａｐｈｙｌｌｕｍ（Ｔｈｕｎｂ）Ｍａｋ,为葫芦科多年生草质藤本,又名七叶胆。全世界绞股蓝属植物约有１３种,中国有１３种,其中７种为我国所特有...     

绞股蓝多糖提取分离、化学结构及生物活性的研究进展

绞股蓝是我国特色天然资源,近年来研究发现多糖是其重要的药用活性成分,具有抗肿瘤、免疫调节、抗氧化、降血糖、抗疲劳等生物活性。更重要的是绞股蓝多糖不仅具有显著的抗肿瘤活性,还参与和介导了免疫功能的调节,能够提高机体免疫功能而对正常细胞没有毒副作用,在恶性肿瘤、艾滋病等威胁人类健康疾病的防治和治疗方面具有广阔的应用前景,有希望开发成我国特色的创新药物。本文对绞股蓝多糖分离纯化、结构以及生物活性等方面进行详细综述,为以后进一步研究提供理论参考。     

莫将乌蔹莓错当绞股蓝

20世纪70年，日本学者从绞股蓝中分离出84种皂甙，其中有6种与人参皂甙成分相同，还有多种绞股蓝皂甙水解产生的次级甙、甙元和次级甙元也与人参水解产物相同，由此曾被人们誉为“南方人参”。另据研究，绞股蓝不但具有抗肿瘤、抗疲劳、抗衰老、调节脂质代谢、降低过氧化脂质、镇静、催眠、镇痛、美润肌肤之功效。而且还是一种治疗咳嗽、痰喘、慢性支气管炎以及传染性肝炎的民间中草药。绞股蓝（Gynostemma pentaphyllum)又名七叶胆，是葫芦科多年生攀援草本植物，它分布于日本、越南、印度和印度尼西1亚，以及我国陕西南部、长江流域以南…     

绞股蓝磷脂成分的分析

绞股蓝为胡芦科植物绞股蓝(Gynostemma pentaphyllum Makino的全草,我国民间用于治疗咳嗽、痰喘、慢性气管炎及传染性肝炎等疾病。1983年以来,从绞股蓝全草中提取得到50多种与人参皂甙有类似骨架的达玛烷型皂甙,尚含黄酮、糖类等成分。药理实验表明绞股蓝具有抑制肿瘤细胞繁殖,抗溃疡,抗疲劳,降血脂及延长寿命等作用。为进一步探讨绞股蓝这些药理作用的物质基础,本文首次对绞股蓝磷脂成分进行分析研究。     

绞股蓝、绞股兰及锥形果是三种不同的植物

绞股蓝为葫芦科(cucurbitaceae)植物,具有清热解毒、止咳化痰的作用,对慢性支气管炎、传染性肝炎及肿瘤等疾病有较好的疗效。近年来日本学者先后报导绞股蓝含有多种人参皂甙成分,引起各国有关学者普遍给予重视。国内一些学者也译文介绍,有的还对绞股蓝进行栽培实验。但是一些译文或著述把绞股蓝称为“绞股兰”,有的则用“锥形果”的名称。经考证,绞股蓝并没有“绞股兰”、     

八大公山自然保护区绞股蓝伴生群落特征研究

对八大公山国家级自然保护区的绞股蓝伴生群落特征进行了研究.结果表明,异叶榕、尾叶山茶和水马桑等是常见的对绞股蓝具有遮荫作用的乔灌层植物.绞股蓝伴生群落计有47科86属102种维管植物,以蓼科、伞形科、菊科和百合科种类较多,绵毛金腰是伴生群落的优势种,它与冷水花、吉祥草、楼梯草和箬竹等是绞股蓝的重要伴生种.伴生群落的物种多样性指数表明整个环境适于绞股蓝的生长.     

绞股蓝提取物对小鼠运动能力的影响

目的：研究绞股蓝提取物对小鼠运动能力的影响。方法：建立动物训练实验模型,测定小鼠肝组织的超氧化物歧化酶（SOD）的活性、丙二醛（MDA）的活性。结果：灌服海南野生绞股蓝提取液后,服药组小鼠肝组织SOD活性显著高于各自对照组．MDA含量显著低于各自对照组。结论：海南野生绞股蓝具有较强的抗脂质过氧化损伤的作用,服用其绞股蓝可消除运动过程中产生过量自由基,保护心脏、肝脏免受自由基的损伤,对肝脏和心肌组织具有显著的保护作用。     

广西绞股蓝属的生态分布及资源利用

本文报道了广西绞股蓝属的种类调查研究概况,并分析了广西绞股蓝属的生态条件和地理分布,为绞股蓝属的资源开发利用提供参考。本属在广西有5种,即广西绞股蓝、扁果绞股蓝、光叶绞股蓝、长梗绞股蓝。     

Effects of temperature on growth and total gypenosides accumulation in Gynostemma pentaphyllum and Gynostemma pentagynum

温度是影响绞股蓝生长发育和总皂苷积累的重要环境因子之一.将绞股蓝和五柱绞股蓝幼苗置于10、15、20、25 ℃和30 ℃的光照培养箱中处理40 d,检测其形态指标和总皂苷含量.结果表明:在25 ℃条件下,绞股蓝的叶面积、叶柄长、茎长、新萌叶片数、生物量和总叶绿素含量均为最高,五柱绞股蓝的生长发育也具有类似的规律,因此推断25 ℃是绞股蓝和五柱绞股蓝生长发育的适温条件.绞股蓝和五柱绞股蓝的总皂苷含量则以30 ℃下最高.绞股蓝的生物量和总皂苷含量决定了总皂苷产量,25～30 ℃最有利于提高绞股蓝的总皂苷产量,30 ℃则是提高五柱绞股蓝总皂苷产量的最适温度.     

广西绞股蓝属的生态分布及资源利用

本文报道了广西绞股蓝属的种类调查研究概况,并分析了广西纹股蓝属的生态条件和地理分布,为绞股蓝属的资源开发利用提供参考。本属在广西有5种,即广西绞股蓝、扁果绞股蓝、光叶绞股蓝、长梗绞股蓝。     

不同碳氮源对糙皮侧耳木质纤维素酶活性的影响

以糙皮侧耳Pleurotus ostreatus为材料,研究简单碳氮源及木质素纯品诱导条件对其木质纤维素酶活性的影响。结果表明,不同的碳源培养基和氮源培养基对糙皮侧耳漆酶活性、羧甲基纤维素酶活性和木聚糖酶活性均具有极显著的影响（P<0.001）,且对糙皮侧耳菌丝生物量也有极显著的影响（P<0.001）。以蔗糖作主要碳源诱导物时,有利于提高糙皮侧耳漆酶活性;以果糖作主要碳源诱导物时,有利于提高糙皮侧耳羧甲基纤维素酶活性和菌丝生物量的积累;以葡萄糖作主要碳源诱导物时,有利于提高糙皮侧耳木聚糖酶活性。以酵母浸粉作主要氮源诱导物时,有利于提高糙皮侧耳漆酶活性和菌丝生物量的积累;以硝酸钾作为主要氮源诱导物时,有利于提高糙皮侧耳羧甲基纤维素酶活性;以硫酸铵作为主要氮源诱导物时,有利于提高糙皮侧耳木聚糖酶活性。碱性木素的存在,有利于提高糙皮侧耳漆酶活性,但不利于菌丝生物量的积累。与此同时,碱性木素的存在对糙皮侧耳羧甲基纤维素酶和木聚糖酶活性并没有促进作用。     

Regulation of rat-liver glycoprotein: N-acetylneuraminic acid transferase activity by pyrimidine nucleotides.

CMP-N-acetylneuraminic acid: glycoprotein sialyltransferase activities were assayed in rat liver microsomal fractions using desialylated fetuin as the substrate acceptors for N-acetylneuraminic acid. It was found that cytidine nucleotides specifically depressed enzyme activities. CMP was shown to act as a competitive inhibitor with an apparent Ki of 0.62 mM. N-Acetylneuraminic acid at 1.15 mM had no effect on enzyme activities. Uridine nucleotides at 1.15 mM, especially UDP, increased enzyme activities. UDP may act as an allosteric activating agent increasing the apparent V. Other nucleotides, sugars and nucleotide-sugars at similar concentrations affected sialyltransferase activities only slightly. A general mechanism is proposed for the regulation of glycosyltransferase activities by free nucleotides.     

The use of biological activities to monitor the removal of fuel contaminants—perspective for monitoring hydrocarbon contamination: a review

Soil biological activities are vital for the restoration of soil contaminated with hydrocarbons. Their role includes the biotransformation of petroleum compounds into harmless compounds. In this paper, the use of biological activities as potential monitoring tools or bioindicators during bioremediation of hydrocarbon-contaminated soil are reviewed. The use of biological activities as bioindicators of hydrocarbon removal in soil has been reported with variable success. This variability can be attributed partially to the spatial variability of soil properties, which undoubtedly plays a role in the exposure of organisms to contaminants. Widely used bioindicators have been enzyme activities, seed germination, earthworm survival and microorganisms or microbial bioluminescence. A mixture of some successful utilization of biological activities and several failures, and inconsistencies reported, show that at this stage there is no general guarantee of successful utilization of biological activities as monitoring tools. Wherever possible, the use of biological activities as bioindicators of hydrocarbon removal must be used to complement existing traditional monitoring tools.     

Biological activities of analogues of lipid A based chemically on the revised structural model. Comparison of mediator-inducing, immunomodulating and endotoxic activities

Lipid A analogues were chemically synthesized based on the model structure recently revised, and biological activities of the analogues were tested. The analogue, (beta-1,6)-linked glucosamine disaccharide carrying ester-bound 3-hydroxytetradecanoic acids at 3 and 3' position of reducing and nonreducing glucosamine in addition to amide-bound 3-hydroxytetradecanoic acids and glycosidic-linked and ester-linked phosphate groups, showed much stronger activities for mediator inducing and immunomodulating as well as endotoxic activities than those exhibited by the previously synthesized analogues based on the old model. Among the activities tested, induction of interferon and tumor necrosis factor as well as mitogenicity, adjuvanticity and pyrogenicity were, however, not expressed so strongly as natural lipid A used as controls. In contrast, the analogue exhibited comparable activities to those of control lipid A in the test of lethal toxicity to mice and gelating activity of Limulus amebocyte lysate. Other synthetic analogues carrying a phosphate group showed comparable, slightly stronger or weaker activities depending on the test, but nonphosphorylated analogue exhibited no apparent or only very weak activities.     

Effects of backbone structures and stereospecificities of lipid A-subunit analogues on their biological activities

Among chemically synthesized analogues corresponding to the nonreducing sugar part of lipid A, we have found an analogue (GLA-27) which exhibits Limulus, mitogenic, polyclonal B cell activation (PBA), interferon-inducing, and tumor necrosis factor (TNF)-inducing activities but not pyrogenic activity. The structure of GLA-27 comprises 4-O-phosphono-D-glucosamine with tetradecanoyl and 3-tetradecanoyloxytetradecanoyl (C14-O-(C14] groups as the 3-O- and 2-N-acyl substituents, respectively. Derivatives of GLA-27 with different backbone structures, such as the 1-deoxy, 3-epimeric, 3-amino, and 1-deoxy-3-epimeric derivatives of glucosamine, were chemically synthesized, and their mediator-inducing activities such as interferon- and TNF-inducing activities were investigated in comparison with their B cell activation activities including mitogenic and PBA activities. Among these derivatives, a derivative with a 1-deoxyglucosamine backbone (GLA-40) exhibited stronger B cell activation activities than those of GLA-27 while the mediator-inducing activities of GLA-40 were weaker than those of GLA-27. In addition to these derivatives, stereoisomers of GLA-27 which possess the (R) and (S) forms of C14-O-(C14) as the 2-N-acyl substituent were also synthesized and their biological activities compared. The (S) isomer exhibited much stronger mediator-inducing activities than the (R) isomer. On the other hand, B cell activation activities of the (R) isomer were strong and those of the (S) isomer weak. These results clearly demonstrate that mediator-inducing activities and B cell activation activities can be selectively expressed by modifying the structures of lipid A analogues.     

Comparative distribution of glutamyl and aspartyl aminopeptidase activities in mouse organs.

To evaluate the functional role of glutamyl and aspartyl aminopeptidases, their soluble and membrane-bound activities were measured simultaneously in several tissues of normal mice using arylamide derivatives as substrates. Although the soluble aspartyl aminopeptidase activity showed its highest levels in the testicle, the rest of the activities presented their highest levels in the kidney. Different patterns of distribution were observed for glutamyl and aspartyl aminopeptidase activities and also for soluble and membrane-bound aspartyl aminopeptidase activities. However no major differences were observed between soluble and membrane-bound glutamyl aminopeptidase activities. This unequal distribution suggests that the use of arylamide derivatives as substrates is a sensitive method that distinguishes between these enzymatic activities. The results also suggest different functions for soluble and membrane-bound aspartyl aminopeptidase activities, and for glutamyl and aspartyl aminopeptidase activities.     

Significant differences in the activities of alpha-amylases in the absence and presence of polyethylene glycol assayed on eight starches solubilized by two methods

Starch is a reserve chemical source of the energy of the sun found in plants as a water-insoluble granule that differs in their chemical and physical properties, depending on the source. The granules can be solubilized by heating in water or by treatment with various reagents, such as 1M NaOH. alpha-Amylases are widely distributed enzymes that initiate the hydrolysis of starch into low molecular weight maltodextrins. We recently found that the activities of a single alpha-amylase on two different starches were significantly different. We then determined the activities of Bacillus amyloliquefaciens and porcine pancreas alpha-amylases, using eight different starches, solubilized by two methods: autoclaving at 121 degrees C and 1M NaOH at 20 degrees C. There were significant differences in the activities of both of the amylases on all eight of the starches. Previously, it had been found that polyethylene glycol (PEG) stabilized and activated the activities of both enzymes, using a soluble amylose as the substrate. Addition of PEG to the enzymes greatly increased the activities on the eight starches, but the activities still differed significantly. The different activities with the starches were hypothesized as differences in the amounts of secondary and tertiary structures that are partially retained when the different starches are solubilized; the activities on addition of PEG is hypothesized as the formation of highly active species from a series of less active forms.     

Drug induction of hepatic glutathione S-transferases in male and female rats*.

The induction of the glutathione S-transferases by phenobarbital and polycyclic hydrocarbons was studied in male and female rats. Administration of phenobarbital resulted in 60-80% increase in S-aryl and S-aralkyl enzyme specific activities, whereas the S-epoxide and S-alkyl activities were increased by 30-40%. In following the sequence of induction, the former two activities were noted to reach peak activities before an increase in the latter two activities was observed. Both 3-methylcholanthrene and 3,4-benzopyrene were shown toi nduce these four enzymic activities, although without the discrimination between pairs of activities noted with phenobarbital. No change in Km accompanied the increase in Vmax. after induction by drugs, and no change occurred in Ki for sulphobromophthalein inhibition. Significantly lower enzyme specific activities were found for three of the activities studied in female rats but no difference was observed in the S-alkyltransferase activity. However, the proportional increase in the enzymic activities in response to phenobarbital was the same in males and females. These studies demonstrate the drug induction of a group of cytosolic drug-metabolizing enzymes as well as the identification of sex differences in these activities.     

Basidiomycetes harbour a hidden treasure of proteolytic diversity

This survey is the first to investigate the proteolytic potential of a large number of basidiomycetes. Aqueous extracts of 43 basidiomycetes were investigated for their content of proteolytic activities, using gelatin zymography. The activities were characterised qualitatively using class specific inhibitors. All four catalytic classes of proteases were present, with 4% of all activities classified as aspartic, 5% as cysteine, 6% as metallo and 22% as serine proteases, while the remaining activities could not be assigned unambiguously. The majority of the latter were not inhibited by any of the inhibitors used and were termed insensitive. Different proteolytic activities are evenly distributed among members of all orders of basidiomycetes, although some taxa are a richer source of proteases than others. A significant number of the cysteine protease activities shown here have not previously been reported in basidiomycetes. The fungal cysteine and serine protease inhibitors, clitocypin and CNSPI (Clitocybe nebularis serine protease inhibitor), both inhibited a number of activities and even a few activities that were otherwise insensitive to all other inhibitors used, hence indicating their potential for a regulatory role. The number and diversity of proteases in basidiomycetes are seen to be remarkable and encourage further investigation.     

Characterization of endothelin converting enzyme activities in soluble fraction of bovine cultured endothelial cells

Endothelin converting enzyme activities in the soluble fraction of cultured bovine aortic endothelial cells were characterized. The two major endothelin converting enzyme activities were eluted from a hydrophobic chromatography column and the elution profile of the endothelin converting enzyme activities was the same as that of cathepsin D activities. These activities had a same pH optimum at pH 3.5 and were effectively inhibited by pepstatin A. Furthermore, anti-cathepsin D antiserum absorbed these activities as well as cathepsin D activity. Immunoblotting analysis using the antiserum showed the major active fractions have immunostainable components of identical molecular weights with cathepsin D. From these results, we concluded that the major endothelin converting activities in the soluble fraction of endothelial cells are due to cathepsin D. In addition to these cathepsin D activities, a minor endothelin converting enzyme activity with an optimum pH at 3.5 was found, which does not have angiotensin I generating (cathepsin D) activity from renin substrate and needs much higher concentrations of pepstatin A to inhibit the activity than cathepsin D.