中药独活的化学成分及其抗肿瘤活性的研究进展

独活在临床治疗中运用广泛。现代研究表明独活含有多种抗肿瘤活性成分,包括甲氧基欧芹素、补骨脂素、花椒毒素、香柑内酯、伞形花内酯、异欧前胡素。本文就近年来对独活的化学成分及其抗肿瘤作用研究进展情况进行综述。     

中药独活的化学成分及其抗肿瘤活性的研究进展

独活在临床治疗中运用广泛。现代研究表明独活含有多种抗肿瘤活性成分,包括甲氧基欧芹素、补骨脂素、花椒毒素、香柑内酯、伞形花内酯、异欧前胡素。本文就近年来对独活的化学成分及其抗肿瘤作用研究进展情况进行综述。     

中药蛇床子水溶性提取物中化学成分的诱变性研究

本文采用Ａｍｅｓ、活体小鼠骨髓细胞染色体畸变和骨髓多染红细胞微核试验对中药蛇床子水溶性提取物中９种化合物进行了诱变性研究。结果表明，这９种化合物既无移码突变和碱基换效应，亦无诱发小鼠骨髓细胞染色体损伤和骨髓细胞抑制作用。     

天然蛇床子素的抗肿瘤活性实验研究

背景与目的:蛇床子素是天然存在的香豆素化合物,本研究旨在检测天然蛇床子素在小鼠体外和体内的抗肿瘤活性.材料与方法:体外蛇床子素对人肺腺癌细胞A549和人肝癌细胞Bel-7402的抗肿瘤实验采用MTF法,并计算IC50;体内蛇床子素对小鼠肝癌H::实体瘤的抗肿瘤实验采用常规的抗肿瘤实验方法,用昆明种小鼠,雌雄各半,设空白对照组、顺铂(5 mg/kg)和香菇多糖(1 mg/kg)2个阳性对照组和1.11、1.67、2.50 mg/kg 3个剂量蛇床子素给药组,每组12只小鼠;用蛇床子素给小鼠灌胃后观察抑瘤率和胸腺、脾指数及肝重量变化,采用t检验进行数据的统计学处理.结果:蛇床子素体外对肺腺癌细胞A549和人肝癌细胞Bel-7402的半数抑制浓度IC50分别为:67.83、123.92 μg/ml;体内对小鼠肝癌H22实体瘤抑瘤率达62%～73%,各给药组与空白对照组比较差异均具有统计学意义(P＜0.01),脏器指数及重量与空白对照组比较差异均无统计学意义(P＞0.05),而与阳性对照顺铂组间的差异具有统计学意义(P＜0.01).结论:蛇床子素体外和体内对实验肿瘤均有明显的抗肿瘤活性,而且在给药剂量下实验动物未出现任何毒性反应.     

佛手柑内酯对鼻咽癌细胞凋亡的影响北大核心CSCD

目的探讨佛手柑内酯对鼻咽癌细胞凋亡的影响及可能的作用机制。方法 CCK-8法检测佛手柑内酯对鼻咽癌细胞CNE-2和HONE-1体外活性的抑制作用。Annexin V-FITC/PI双染法、JC-1染料法分别检测细胞的凋亡率及线粒体膜电位,并检测Caspase-3的活性水平。实时定量RT-PCR与Western blot检测BAX与BCL2基因的mRNA水平和蛋白表达量,并检测STAT3蛋白表达量及其Y705位点的磷酸化水平。结果在10μM和100μM两个浓度佛手柑内酯作用下,CNE-2和HONE-1细胞凋亡率及Caspase-3活性明显上升(P<0.05),线粒体膜电位明显下降(P<0.05);BCL2基因的mRNA及蛋白表达水平降低,BAX基因的mRNA及蛋白表达水平升高,STAT3蛋白表达量不变而其Y705位点的磷酸化水平下降。结论佛手柑内酯能破坏鼻咽癌细胞线粒体功能并诱导其发生凋亡,这一过程可能与下调BCL2基因、上调BAX基因及抑制STAT3磷酸化有关。     

沙棘油抗突变作用的实验研究

自然环境中存在多种致突变致畸，致癌物质，人们发掘它们．用以预防和治疗由此所引发的不良效应，这已成为国内外学者关注的热点．植物中主要的抗诱变成分有多酚、芳香异硫氨酸盐，香豆素．黄酮，双萜类、维生素A类、维生素C．α-生育酚，硒盐和植物固醇类等．沙棘属于胡类子科沙棘属植物，含有多种生物活性物质，它能提高机体免疫力的效应已为实验所证实．     

蛇床子素对肺腺癌、肺鳞癌生长抑制作用的实验研究

目的: 观察蛇床子素对人肺鳞癌和肺腺癌的抑制作用。方法:建立BALB/ C 裸鼠的人肺腺癌和肺鳞癌模型,给予蛇床子素,剂量为1. 5μg/ (g·d) ,观察瘤体的大小、重量和动物血清中肺癌标志物DR270 的水平,以此评价蛇床子素的抑癌作用。结果:蛇床子素对肺鳞癌的抑瘤率为69. 5 % ,对肺腺癌的抑瘤率为50. 0 % ,对DR270 也有显著降低作用。结论:蛇床子素对肺鳞癌和肺腺癌的瘤体生长有一定的抑制作用,尤其是肺鳞癌。     

中药独活及其单体蛇床子素体外抗血管生成的实验研究

目的：探讨独活醇提物及其单体蛇床子素对体外血管生成的抑制作用及其可能的机制.方法：采用MTT法观察中药独活醇提物及蛇床子素对人脐静脉血管内皮细胞（human umbilical vein endothelial cell.HUVEC）和人肠癌LoVo细胞增殖的影响,Transwell小室趋化实验、体外小管形成实验以及流式细胞术,观察并比较独活醇提物及蛇床子素对HUVEC迁移、小管形成、凋亡及周期的影响.结果：3.75-30μg/ml的独活醇提物及蛇床子素作用48h时对HUVEC的细胞增殖抑制率分别在5.16％-10.15％和22.64％-65.56％之间,对LoVo细胞增殖抑制率分别在2.86％-7.29％和5.15％-24.39％之间.体外小管及小管迁移实验显示,3.75-30μg/ml的蛇床子素作用24h时HUVEC小管形成数目减少,且管腔不完整.3.75-30μg/ml的独活醇提物和蛇床子素处理12h对HUVEC迁移抑制率分别在-2.16％至8.00％和13.70％至63.04％之间.3.75-30μg/ml的独活醇提物和蛇床子素诱导HUVEC细胞凋亡率分别在6.1％-14.4％和18.8％-89.5％之间.独活醇提物和蛇床子素作用HUVEC 24h后,使内皮细胞周期主要阻滞在G0-G1期,蛇床子素对细胞周期影响强于独活醇提物.结论：蛇床子素在体外抑制血管生成作用强于独活醇提物,说明蛇床子素可能是独活醇提物中发挥抗血管生成作用的主要成分,其作用机制可能与抑制HUVEC增殖、迁移和小管形成,诱导HUVEC凋亡,阻滞HUVEC细胞周期有关.     

调味品花椒的诱变性研究

花椒是人们常用的调味品,鉴于其与人民生活关系密切,应慎重对待。故我们用果蝇伴性隐性致死(SLRL)小鼠骨髓多染性红细胞(PCE)微核和精子畸形等测试作进一步的验证。实验结果表明:花椒对果蝇SLRL的总诱变率为0.2％,低于自发突变率0.4％     

独活醇提物及其单体蛇床子素体外抗肿瘤活性的实验研究

目的：探讨独活醇提物及其单体蛇床子素在体外对人胃癌细胞株MKN-45、BGC-823、人肺腺癌细胞株A549、人乳腺癌细胞株MCF-7及人结肠癌细胞株LOVO生长的影响.方法：将不同浓度中药独活醇提物及其单体蛇床子素作用于人胃癌细胞株MKN-45、BGC-823、人肺腺癌细胞株A549、人乳腺癌细胞株MCF-7及人结肠癌细胞株LOVO,倒置显微镜下观察细胞形态变化,MTT法测定独活醇提物及其单体蛇床子素对5株肿瘤细胞的增殖抑制率.结果：蛇床子素浓度在3.75μg/ml-120 μg/ml之间对MKN-45、BGC-823、A549、MCF-7及LOVO均有明显的抑制作用,且呈剂量-效应关系,IC50值分别为43.299μg/ml、56.790μg/ml、37.444 μg/ml、38.397μg/ml、36.231 μg/ml;相同条件下,独活醇提物对5株肿瘤细胞的抑制作用都较弱.结论：蛇床子素在体外能够抑制MKN-45、BGC-823、A549、MCF-7和LOVO的增殖,且LOVO细胞对蛇床子素最敏感,蛇床子素对5株肿瘤细胞的抑制作用均强于独活醇提物,说明蛇床子素有可能是独活醇提物中抗肿瘤作用的主要成分.     

Oral administration of naturally occurring coumarins leads to altered phase I and II enzyme activities and reduced DNA adduct formation by polycyclic aromatic hydrocarbons in various tissues of SENCAR mice

Several naturally occurring coumarins, to which humans are routinely exposed in the diet, were previously found to inhibit P450-mediated metabolism of benzo[a]pyrene (B[a]P) and 7,12-dimethylbenz[a]anthracene (DMBA) in vitro, block DNA adduct formation in mouse epidermis and inhibit skin tumor initiation by B[a]P and/or DMBA when applied topically to mice. The present study was designed to investigate the effects of two of these compounds, of the linear furanocoumarin type, when given orally (70 mg/kg per os, four successive daily doses), on P450 and glutathione S-transferase (GST) activities and DNA adduct formation by B[a]P and DMBA in various mouse tissues. Imperatorin and isopimpinellin significantly blocked ethoxyresorufin O-deethylase (EROD) and pentoxyresorufin O:-dealkylase (PROD) activities in epidermis at 1 and 24 h after oral dosing. Imperatorin and isopimpinellin modestly inhibited EROD activities in lung and forestomach at 1 h and significantly inhibited PROD activities in lung and forestomach at 1 h after the final oral dose. Twenty-four hours after the final oral dose of imperatorin or isopimpinellin EROD and PROD activities remained inhibited in epidermis and lung. However, forestomach P450 activity had returned to control levels. Interestingly, imperatorin and isopimpinellin treatment inhibited liver EROD activity at 1 h, had no effect on PROD activity at this time point, but elevated both these enzyme activities at 24 h. Elevated EROD and PROD activities coincided with elevated hepatic P450 content. Imperatorin and isopimpinellin treatment also increased liver cytosolic GST activity at both 1 and 24 h after the final oral dose by 1.6-fold compared with corn oil controls. Oral administration of imperatorin and isopimpinellin also had a protective effect against DNA adduct formation by B[a]P and DMBA. Imperatorin pretreatment decreased formation of DNA adducts by DMBA in forestomach. Pretreatment with isopimpinellin led to reduced DNA adduct levels in liver (B[a]P), lung (B[a]P) and mammary epithelial cells (DMBA). These results suggest that imperatorin and isopimpinellin may have potential chemopreventive effects when administered in the diet.     

Oral administration of the citrus coumarin,isopimpinellin, blocks DNA adduct formation and skin tumor initiation by 7,12-dimethylbenz[a]anthracene in SENCAR mice

The current study was designed to evaluate the effects of oral administration of the citrus coumarin, isopimpinellin, on skin tumor initiation by topically applied benzo[a]pyrene (B[a]P) and 7,12-dimethylbenz[a]anthracene (DMBA). To evaluate the effects of orally administered isopimpinellin on skin tumor initiation by B[a]P and DMBA, its effects on DNA adduct formation were first evaluated. Female SENCAR mice were pre-treated twice with corn oil, or isopimpinellin (70 mg/kg body wt per os) at 24 h and 2 h prior to topical treatment with B[a]P or DMBA. Another citrus coumarin, imperatorin, was also included in these experiments for comparison. Orally administered isopimpinellin and imperatorin significantly inhibited B[a]P-DNA adduct formation by 37 and 26%, respectively. Imperatorin also blocked DMBA-DNA adduct formation by 43%. In a second dose-response study, orally administered isopimpinellin (35, 70 and 150 mg/kg) blocked DMBA-DNA adduct formation by 23, 56 and 69%, respectively. For the tumor study, mice were pretreated orally with corn oil or isopimpinellin at 24 and 2 h prior to initiation with DMBA, and 2 weeks later promotion began with 12-O-tetradecanoylphorbol-13-acetate (TPA). Isopimpinellin significantly reduced the mean number of papillomas per mouse by 49, 73 and 78% compared to corn oil controls at 30, 70 and 150 mg/kg body wt, respectively. Orally administered isopimpinellin also significantly reduced the percentage of mice with papillomas at the highest dose tested (150 mg/kg). The effectiveness of isopimpinellin given topically over a broad dose range against DMBA tumor initiation was also evaluated for comparison. As part of this study, several parameters of systemic toxicity were evaluated following oral dosing with isopimpinellin and imperatorin. Mice were treated orally with corn oil, isopimpinellin or imperatorin (35, 70 and 150 mg/kg body wt per os) once daily for four consecutive days, killed at 24 h after the last dose, and livers, lungs, and kidneys evaluated histologically. In addition, urinary parameters of nephrotoxicity, blood parameters of liver and kidney function, and thrombin clotting time were assayed. No significant changes in blood clotting, or renal or hepatic function were observed. There was, however, a significant increase in liver wt accompanied by cytoplasmic vacuolation of hepatocytes. There were no histopathological changes in lungs or kidneys. Overall, these data indicate that isopimpinellin (and imperatorin) have chemopreventive effects when administered orally on skin tumor initiation by DMBA.     

Naturally occurring coumarins inhibit 7,12-dimethylbenz[a]anthracene DNA adduct formation in mouse mammary gland

Naturally occurring coumarins (NOCs) are anti-carcinogenic in the mouse skin model. To characterize the chemopreventive potential of NOCs against breast cancer, we first examined their effects on 7,12-dimethylbenz[a]anthracene (DMBA)-DNA adduct formation in mouse mammary gland. We hypothesized that those NOCs that both inhibited cytochrome P450 1A1/1B1 and induced hepatic glutathione S-transferases (GSTs) would be the most effective in blocking DMBA-DNA adduct formation in mouse mammary gland. To address this hypothesis, simple coumarins (e.g. coumarin and limettin, which induced mouse hepatic GSTs but had little effect on P4501A1/1B1) and linear furanocoumarins (e.g. imperatorin and isopimpinellin, which induced hepatic GSTs and were potent inhibitors of P4501A1/1B1) were compared. Mice were pretreated with NOCs (150 mg/kg body wt, by gavage) prior to either a single dose of DMBA (50 microg) or multiple doses of DMBA (20 microg daily for 3 and 6 weeks). Mammary DMBA-DNA adduct formation was quantitated by the nuclease P1-enhanced 32P-postlabeling assay. With the single dose of DMBA, coumarin, limettin, imperatorin and isopimpinellin inhibited DMBA-DNA adduct formation by 50, 41, 79 and 88%, respectively. Coumarin, limettin and imperatorin blocked DMBA-DNA adduct formation by 36, 60, and 66% at 3 weeks, and by 0, 49 and 55% at 6 weeks of DMBA dosing, respectively. In a 6 week dose-response study of select NOCs and 7,8-benzoflavone (a potent P4501 inhibitor that had little effect on GSTs), DMBA-DNA adduct formation was inhibited by 0, 43 and 24% in the limettin groups; by 26, 26 and 69% in the isopimpinellin groups; and by 80, 96 and 97% in the 7,8- benzoflavone groups at 35, 70 and 150 mg/kg, respectively. Taken together, these results suggest that linear furanocoumarins had a greater inhibitory effect on DMBA-DNA adduct formation in mouse mammary glands compared with simple coumarins, and that the predominant effect may be P4501 inhibition.     

Antimutagenicity of selenium-enriched rice on mice exposure to cyclophosphamide and mitomycin C

The in vivo antimutagenicity of Se-enriched rice was evaluated by bone marrow micronucleus and testicle chromosomal aberrations assay in mice exposed to cyclophosphamide (CP) and mitomycin C (MMC). Regular rice did not alter the occurrence of chemical-induced mutation. However, the addition of Se-enriched rice or selenite significantly inhibited the incidence of CP-induced micronuclei and MMC-induced chromosomal aberration in mice and the effect was dose-dependent. Providing selenite or Se-enriched rice also significantly increased the activity of glutathione peroxidase in liver and the selenium concentration in blood compared to regular rice. No significant differences were found in mice body weight gain. These results revealed the antimutagenic potential of Se-enriched rice against chemical-induced mutation.     

有机稀土化合物对SOS反应的抑制作用①

本文应用ＳＯＳ显色法研究了三种有机稀土化合物及其相应鳌合剂的抗诱变作用。抗坏血酸稀土、柠檬酸稀土及乙二胺四乙酸稀土浓度分别在０．５～２．５ｍｇ／ｍｌ、１～２Ｏｍｇ／ｍｌ及５～４Ｏｍｇ／ｍｌ时，对已知致癌物亚硝基胍和３，４—苯并芘诱导的ＳＯＳ反应具有抑制作用，且有较好的剂量效应关系。而单纯的螫合剂抗坏血酸抗诱变效果很好，柠檬酸显示微弱的抗诱变性，乙二胺四乙酸二钠则无抗诱变作用。     

Chemopreventive Activity of Porphyrin Derivatives Against 6-Sulfooxymethylbenzo[a]pyrene Mutagenicity

Porphyrins exhibit potent antimutagenic activity in the range of assays in vitro and in vivo. The antimutagenic ‍activities of porphyrin derivatives including phthalocyanines (Pcs) were investigated using 6- ‍sulfooxymethylbenzo[a]pyrene (SMBP) and its proximate metabolite 6-hydroxymethylbenzo[a]pyrene (HMBP) in ‍the Ames assay and hypoxanthine:guanine phosphoribosyl transferase (hprt) point mutation assay in V79 cells. Pcs, ‍irrespective of coordinated metal, showed highly antimutagenic activity against both HMBP and SMBP in the Ames ‍assay. However, their inhibitory effects against HMBP were in general less dramatic than against SMBP. Treatment ‍with chlorophyllin (CHL) and protoporphyrin reduced the mutation frequency to 24.8% and 19.1% of that with ‍SMBP, and 56.5% and 40.7% of that with HMBP, respectively. Hemin, biliverdin and chlorophylls had a lower ‍antimutagenic activity compared with other porphyrin derivatives although they still retained inhibitory capacity ‍against SMBP. The results of the Ames test for SMBP were extended to the mammalian system to confirm the ‍antimutagenicity of porphyrin derivatives. The antimutagenic strength of porphyrin derivatives was in order of ‍hemin, Pcs, CHL and protoporphyrin. Biliverdin, chlorophyll a and chlorophyll b had a little effect against HMBP ‍and SMBP. Among the compounds tested here, hemin had a strong inhibitory effect against SMBP-induced mutation ‍in V79 cells while Pcs were most effective in the microbial system. It is assumed that these discrepancies are partly ‍due to differences in membrane permeability to the chemicals, their metabolisms and chromosomal organization. ‍Furthermore, carcinogen-DNA binding in calf thymus DNA and plasmid was carried out with a series of porphyrin ‍derivatives to understand their protective mechanism. Most of the porphyrin derivatives exhibited inhibitory activity ‍against SMBP with a variety of degrees. A significant reduction of SMBP-mediated DNA damage by Pcs was confirmed ‍on agarose electrophoresis. There was little correlation between the levels of SMBP-DNA adducts and their modulation ‍in mutation frequencies, indicating that porphyrin compounds somehow affect the antioxidant function and the ‍reactivity of SMBP to cellular components.     

4 种水果的致突变及抗突变同步试验

 目的　为筛选有抗突变作用的天然水果。方法　采用抗突变和致突变同步快速试验对猕猴桃、石榴籽、冬雪蜜桃及野茄果 4种可食性水果进行了试验。做加和不加大鼠肝脏微粒体酶(S9)的两种试验。结果　全部受试物均未发现致突变毒性 ;猕猴桃及石榴籽显示对丝裂霉素C引起的致突变作用有拮抗效应 ;冬雪蜜桃有间接抗突变作用 ;野茄果未显示抗突变性。结论　猕猴桃、石榴籽及冬雪蜜桃是抗突变剂。     

Evaluation of Anticarcinogenic and Antimutagenic Potential of Bauhinia variegata Extract in Swiss Albino Mice

Background: Infusions from the bark of Bauhinia is used to treat various diseases in the traditional medicalsystem of India and decoction of the roots is used in dyspepsia and act as an antidote to snake poison. Itschemopreventive potential for cancer was the subject of the present study. Materials and methods: To evaluatethe anticarcinogenicity and antimutagenicity of Kachanar extract a skin carcinogenesis and melanoma tumourmodel was used, along with micronucleus and chromosomal aberration tests, in Swiss albino mice. Results: Inthe skin papilloma model, significant prevention, with delayed appearance and reduction in the cumulative no.of papillomas was observed in the DMBA + Kachanar + croton oil treated group as compared to the DMBA +Croton Oil group. C57 Bl mice which received a 50 % methanolic extract of Kachanar extract at the doses of500 and 1000 mg/ kg body weight for 30 days showed increase in life span and tumour size was significantlyreduced as compared to controls. In antimutagenicity studies,a single application of Kachanar extract at dosesof 300, 600 and 900 mg/kg dry weight, 24 hours prior the i.p. administration of cyclophosphamide (at the 50 mg/kg) significantly prevented micronucleus formation and chromosomal aberrations in bone marrow cells ofmice, in a dose dependent manner. Conclusions: Our results suggest that Kachanar extract exerts anticarcinogenicand antimutagenic activity.     

欧前胡素对体外培养的人表皮黑素细胞Rab27a和酪氨酸酶的作用

目的： 观察欧前胡素对体外培养的人表皮黑素细胞转运相关蛋白Rab27a和酪氨酸酶的作用。方法：体外分离、纯化培养人表皮黑素细胞。随机分为空白对照组和欧前胡素处理组,以50或25 μmol/L欧前胡素作用48 h后观察黑素细胞形态,免疫荧光法检测Rab27a 和酪氨酸酶在黑素细胞中的定位及变化,应用透射电镜观察黑素细胞黑素小体的超微结构。结果：欧前胡素作用48 h后,黑素细胞的树状突起缩短,免疫荧光检测显示,黑素细胞Rab27a和酪氨酸酶蛋白在细胞内定位未见明显变化,但欧前胡素作用后Rab27a蛋白的荧光强度轻度增强(P<0.05),而酪氨酸酶荧光强度降低(P<0.05)。透射电镜结果可见黑素细胞中的黑素小体数量减少。结论：欧前胡素对体外培养的人表皮黑素细胞Rab27a表达呈促进作用,对酪氨酸酶活性及黑素合成呈抑制作用。     

蛇床子素对人前列腺癌LNCaP细胞衰老的影响及其机制研究

目的:探讨天然化合物蛇床子素对人前列腺癌LNCaP细胞衰老的影响及分子机制。方法:采用CCK8法检测蛇床子素对LNCaP细胞增殖的影响；用EdU细胞增殖检测试剂盒检测细胞增殖情况；流式细胞仪检测蛇床子素对其周期阻滞情况；β-半乳糖苷酶染色法检测蛇床子素对LNCaP细胞衰老的影响；Western blot检测衰老相关蛋白SKP2、p27的表达情况。结果:CCK8结果显示蛇床子素对LNCaP细胞的增殖有明显的抑制作用，且呈现一定剂量依赖性（P＜0.05）；EdU结果发现药物处理组细胞在荧光电镜下观察增殖细胞数较空白对照组逐渐减少，当蛇床子素浓度为150 μmol/L时，增殖细胞数较空白对照组下降30%（P＜0.01）；流式细胞仪检测结果显示当蛇床子素浓度为100、150 μmol/L时，阻滞于G2/M期的细胞数明显增多，并且呈现一定的浓度依赖（P＜0.05）；药物处理组细胞于倒置显微镜观察可见，蛇床子素浓度为150 μmol/L时，β-半乳糖苷酶阳性染色细胞数较对照组明显增加（P＜0.01）；免疫印迹法结果显示随着蛇床子素浓度的增加，衰老相关蛋白SKP2的表达减少、p27的表达增加（P＜0.05）。结论:蛇床子素可以抑制前列腺癌LNCaP细胞增殖并进一步诱导其衰老。