Activity Determination of 8 Chinese Herbs against Hepatoma Cell SMMC-7721 in Vitro by MTT Method  

MTT法测定8种中草药体外抗肝癌细胞SMMC-7721活性(摘要)(英文)

作　　者：林董[1] 何爱明[1] 吴丽萍[2] 吴祖建[3] 

机构地区：[1]福建师范大学福清分校生化系,福建福清350300 [2]南昌大学生命科学学院,江西南昌330031 [3]福建农林大学植物保护学院,福建福州350001

出　　处：《Agricultural Science & Technology》2009年第6期111-113,118,共4页农业科学与技术（英文版）

基　　金：Supported by Class A Project of Fujian Educational Committee(JA08054)~~

摘　　要：[Objective] The aim was to build up a set of efficient and rapid models for laboratory to screen anti-hepatocellular carcinoma active substance in vitro. [ Method] By using MTT method, the activities of anti-hepatocellular carcinoma SMMC-7721 in vitro from Cymbopogon distans, Lobelia chinensis, Buddleja offlcinalis, Glycyrrhiza uralensis, Sanguisorba officinalis, Bupleurum chinense, Apium graveolen and Curuma zedoaria were tested. The growth curve of hepatoma cell was described, and the growth status in different periods were observed by inverted microscope. [ Result] Cells induced by active substance would be condensing, clear brim, which have significant differences from normal SMMC- 7721 cells. The results suggested that ESCG, ESCC, ESCB could inhibit proliferation of SMMC-7721 cells at the concentration of 1.0 -1.5 mg/ml, and the inhibition rate were 51.6%, 48.5%, 52.9% respectively. With the increasing of concentration, the inhibition strengthened. [ Conclusion] MTT method could be used as a basic model for screening important anti-hepatoma.[目的]建立一套适合于实验室的高效、快速的体外抗肝癌活性物质筛选模型。[方法]首先,选用乙醇为溶剂对芸香草、半边莲、蜜蒙花、甘草、地榆、柴胡、旱芹和莪术8种中草药植物材料进行浸提。然后通过MTT法对8种中草药体外抗肝癌细胞SMMC-7721的活性进行初测。试验采用二甲基亚砜(DMSO)与甘氨酸-NaOH缓冲液作为溶剂,消除剩余培养基对吸光值的影响。为确保细胞株最大吸收峰位置不变,酶标仪采用570 nm波长测定OD值。为验证筛选出的3种物质对癌细胞的抑制作用,以活性物质浓度为横坐标,细胞生长抑制率为纵坐标,描绘肿瘤细胞生长曲线,倒置显微镜观察细胞不同阶段的生长状况。[结果]倒置显微镜下观察3种细胞阶段的形态:正常状态下,细胞的透明度大、折光性强、轮廓不清;当细胞受污染时,折光性变弱,胞质中常出现空泡、脂滴、颗粒样物质,细胞之间空隙加大,细胞变得不规则,失去原有特点;而受活性物质作用细胞尽管仍然贴壁,但其萎缩,边缘清晰化;三者之间存在一定区别。MTT法培养板定性检测显示,柴胡、莪术、甘草细胞培养孔内的液体颜色都较浅且比较均匀,可见,它们对细胞生长具有明显的抑制作用。而半边莲培养孔内液体颜色深浅不一。还有一些培养孔内液体颜色较深,如卤地菌、蜜蒙花、地榆和旱芹,表明它们对细胞生长没有明显的抑制作用。进一步定量计算表明,浓度为1.0～1.5 mg/ml的甘草乙醇浸提液(ESCG)、莪术乙醇浸提液(ESCC)和柴胡乙醇浸提液(ESCB)对体外培养的肝癌细胞SMMC-7721有较强的抑制增殖作用,抑制率分别为51.6%、48.5%和52.9%。甘草、柴胡和莪术3种物质的抑制作用曲线结果也验证了它们对细胞生长有明显的抑制作用,并且抑制程度与浓度呈正相关。[结论]MTT法筛选可作为重要抗肿瘤的1个基本筛选模型。

关 键 词：MTT method Hepatoma cell SMMC-7721 SCREENING Inhibition rate 

分 类 号：R285[医药卫生—中药学]