地黄活性成分梓醇对转基因CHO细胞M_2受体的调节作用  

Regulatory effect of catalpol from Radix Rehmanniae on M_2 receptor density in M_2 receptor transfected CHO cells

作　　者：王金红[1] 孙启祥[1] 夏宗勤[1] 胡雅儿[1] 

机构地区：[1]上海交通大学医学院细胞调控研究室,上海200025

出　　处：《中国药理学通报》2006年第12期1462-1466,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30371636)

摘　　要：目的探讨地黄活性成分梓醇对转染M2受体亚型基因的CHO细胞系(CHOm2细胞)M2受体密度作用。方法采用衰老CHOm2细胞,常规培养后加入不同浓度梓醇及对照生理盐水,放射配基结合分析测定M2受体密度,Lowry法测定蛋白含量;同时观察了梓醇与M受体结合位点的关系及对乙酰胆碱酯酶的作用。结果梓醇在终浓度10-5、10-4mol.L-1明显升高衰老CHOm2细胞M2受体密度(P<0.01),但不占领M受体结合位点,也不抑制乙酰胆碱酯酶活性(P>0.05)。结论梓醇能上调衰老CHOm2细胞M2受体密度,可能是地黄治疗AD的主要有效成分。Aim To investigate the effect of catalpol from Radix Rehmanniae on M2 receptor density in CHO ceils transfected with gene of M2 receptors （CHO m2）. Methods Cultured CHOm2 ceils were randomly divided into 4 groups： three concentrations of catalpol ： 10^-6,10^-5,10^-4 mol·L^-1 ,and saline control. After addition of catalpol and saline for 72 h, M2 Receptor density was measured by single point 3H-NMS binding assay, the content of protein was measured with Lowry＇s method. Competitive binding assay using the binding system of 3H-NMS was performed to address the question about whether catalpol could occupy the M receptor binding site. Addition of catapol to brain homogenate and measuring the enzyme activity with the Ellman＇s colorimetric method were performed to address the question about whether catalpol could inhibit acetylcholinesterase activity. Results Catalpol can elevate the M2 receptor density in CHO m^2 cells significantly at the doses of 10 ^-5,10^-4 mol · L^-1 （P 〈 0. 01 ） , but not at 10^-6 mol·L^-1. On the other hand, catalpol did not occupy the M receptor binding site, nor did it inhibit the activity of acetylcholinesterase.Conclusion Catalpol has up-regulation effect on M2 Receptor density in a dose-dependent manner, and may be the active component of Radix Rehmanniae in treating diseases with memory impairment.

关 键 词：梓醇 CHOm2细胞 M2受体密度 

分 类 号：R-332[医药卫生] R282.71