Effects of Proliferation of CEF with Ginsenoside and Its Derivatives  

中性红染色法检测人参皂苷及其衍生物对CEF增殖的影响(摘要)(英文)

作　　者：苏建青[1] 褚秀玲[1] 韦旭斌[2] 

机构地区：[1]聊城大学农学院,山东聊城252000 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《Agricultural Science & Technology》2009年第6期97-100,167,共5页农业科学与技术（英文版）

基　　金：Supported by the National Natural Science Foundation of China(No.30471272)~~

摘　　要：Objective The research aimed to provide theoretical basis for studying anti-MDV mechanism of ginsenoside and its derivatives in vitro. Method Effects of ginsenoside and its derivatives on proliferation activity of chick embryo fibroblast （CEF） in vitro were determined by using neutral red dye absorption method. Result The results showed the proliferation effects of different drugs are not completely same, and are more obvious with low toxic drugs. Detected at different action times, the differences of OD values was biggest at 72 h when compared with normal control group, while there was no significant difference at 24 h. Conclusion Ginsenoside and its derivatives could promote the proliferation of CEF cells in medium as low concentrations, which have time-dependent characteristic.[目的]为体外研制人参皂苷及其衍生物的抗MDV作用机制提供理论依据。[方法]分别吸取各试验药物(人参皂苷、人参皂苷衍生物1～8,阳性对照盐酸吗啉胍)溶液0.5 ml至24孔细胞培养板第1孔中,然后1～10孔每孔加入0,5 ml细胞维持液,再将第1孔内液体混匀,吸取0.5ml至第2孔内,再混匀,吸取0.5ml至第3孔内,如此反复至第10孔。则药物的稀释倍数分别为2~1～2^(10)。其浓度分别是3.000 mg/ml,1.500 mg/ml,0.750 mg/ml,0.375 mg/ml,187.500μg/ml,93.750μg/ml,46.880μg/ml,23.440μg/ml,11.720μg/ml,5.860μg/ml。采用中性红染料吸收法,检测人参皂苷及其衍生物对鸡胚成纤维细胞(CEF)体外培养中增殖活性的影响。试验中必须结合细胞病变观察法,通过在光镜下观察细胞的CPE,确定进行测定的最佳时间,以保证结果的准确性。具体操作如下:测定前2 h各孔加入50μl中性红染液,测定时倒掉孔内液体,用PBS洗细胞3次,每孔加入200μl脱色液,置微量振荡器上振荡10 min,使结晶溶解。然后用酶联免疫检测仪测定OD_(570)值,作为细胞增殖的指标。OD值与活细胞的增殖呈正比,OD值越大,表明细胞增殖越旺盛。[结果]人参皂苷在大于750μg/ml浓度下对细胞有毒性,在5.86～46.88μg/ml浓度下对细胞有促增殖作用。毒性作用在72 h下降,而促增殖作用在给药24 h后就表现出来。衍生物1在高浓度的4组均有毒性,OD值显著低于细胞对照组,但衍生物1的促增殖作用不明显。衍生物2在48～72 h分别有5～6组高浓度的OD值显著低于细胞对照组。衍生物3在高于187.5μg/ml的浓度时,OD值低于细胞对照组,并且在72 h时,有2组OD值(11.72和5.86μg/ml)高于细胞对照组,表现出促增殖作用。衍生物4的毒性较高,大部分孔的OD值显著低于正常对照组。衍生物5在浓度高于187.5μg/ml时,OD值低于正常细胞对照组(P<0.05),在24和72 h,分别有一组的OD值显著大于细胞对照组。衍生物6的毒性较�

关 键 词：GINSENOSIDE DERIVATIVES Chicken embryo fibroblast （CEF） Safe concentration 

分 类 号：S853.7[农业科学—临床兽医学]