【作者】 张峰；

【导师】 雷秀玲； 李玛琳；

【作者基本信息】 昆明医学院 ， 药理学， 2004， 硕士

【摘要】 目的：测定一种中药含药血清及五种杀微生物中药制剂抗HIV-1活性，并对含药血清药理学研究方法及药物直接灭活HIV活性的检测方法进行探讨和评价。方法：采用合胞体形成抑制实验检测中药含KDX血清及KDX粗制剂抗HIV-1活性；采用MTT法测定KZL-1、2、3，SSJK及表没食子儿茶素没食子酸酯（EGCG）对人T淋巴细胞系（C8166）的细胞毒性，并以PEG6000的盐溶液浓缩病毒，结合合胞体形成抑制实验检测所选样品对HIV-1的直接灭活作用。结果：（1）未经灭活处理的大鼠含KDX血清空白组、低剂量组、高剂量组及KDX粗制剂抑制半数合胞体形成的浓度(EC₅₀)分别为1／60（v／v），1／106（v／v），1／108（v／v），0.02mg/ml；经灭活处理的各组含KDX血清对HIV-1诱导合胞体形成的抑制率均小于50％，无法计算EC₅₀。（2）KZL-1、2、3，SSJK及EGCG的半数细胞毒性浓度(CC₅₀)分别为1／34（v／V）、1／41（V／V）、1／1698（V／v）、1／633（v／v）、10.27μg／ml；灭活50％HIV-1病毒的浓度(INC₅₀)分别为1／96（v／v）、1／513（v／v）、1／14（v／v）、1／50（v／v）、7.29μg／ml。结论：（1）在所选KDX剂量范围内，大鼠含药血清未表现出抗HIV-1活性，而粗制剂有明显的抗HIV-1活性，EC₅₀为0.02mg/ml。（2）KZL-2对HIV-1有很好的灭活作用，KZL-1次之，而KZL-3、SSJK及EGCG在对HIV-1产生灭活作用的剂量范围内均表现出明显的毒性。（3）中药含药血清与粗制剂实验结果并不总是一致，因而，不能简单地依据中药粗制剂体外实验结果来判断药效。（4）中药含药血清药理学实验更接近体内实验，较直接用粗制剂检测更合理。（5）采用PEG6000的盐溶液浓缩病毒，通过合胞体形成抑制实验检测残留病毒感染性是一种简易、快速、敏感的检测杀微生物侯选物潜在灭活HIV-1活性的实验方法。更多还原

【Abstract】 Objectives: The anti-HIV-1 activity of a kind of drug-containing serum and five kinds of traditional herbal mixtures was detected. Besides, the serologic pharmacologic method and viral inactivation assay were analysed and assessed. Methods: Anti-HIV-1 activity was detected by syncytial formation inhibition assay. KDX direct addition and rats’ KDX-containing serum addition were used respectively. In addition, cytotoxicity of KZL-1,2,3, SSJK, EGCG to Human T-lymphocyte lines(C8166) was detected by MTT assay; PEG6000 solution in saline was used to concentrate viruses. Simultaneously, syncytial formation inhibition assay was used to detect the effect of HIV-1 inactivation. Results: (1) The 50% effective concentration(EC5o) of rats’ KDX-containing sera (not inactivated) control group, low dose group, high dose group and KDX original reagent are l/60(v/v), l/106(v/v), l/108(v/v), 0.02mg/ml, respectively. The 50% effective concentration(EC50) of all groups of rats’ KDX-containing sera (inactivated) are less than 50%, EC650 can not be calculated. (2)The 50% cytotoxic concentration(CC50) of KZL-1,2,3, SSJK and EGCG are l/34(v/v), l/41(v/v), l/1698(v/v), 1/633(v/v) , 10.27μg/ml, respectively; and the 50% HIV-inactivated concentration(INC50) are l/96(v/v),l/513(v/v), l/14(v/v), l/50(v/v), 7.29 (μg/ml, respectively. Conclusions: (l)The KDX original reagent showed obvious anti-HIV activities, EC50 is 0.02mg/ml. However, the rats’ KDX-containing serum didn’t showed anti-HIV activities at the dose adopted in this study. (2)The HIV-1 inactivation effect of KZL-2 was more powerful than KZL-1, and other samples all appeared to obvious cytotoxicity around the HIV-inactivated concentration range. (3)The experiment results of drug-containing serum and original reagent aren’t always consistent,so we can’t draw a conclusion merely according tothe result of the latter. (4)The method of Serologic pharmacologic experiment is more similar to in vivo than that of original reagent direct addition. Apparently, it is more reasonable. (5)PEG6000 solution in saline was used to precipitate viruses, and Syncytial formation inhibition assay was adopted to detect the infection activity of residual viruses,which is a simple, quick and sensitive method to evaluate potential virucidal effect of most microbicide candidates.更多还原