葛根异黄酮抗缺血性脑卒中的活性筛选与研究  

Screening and Study on the Anti-ischemic Stroke Activity of Pueraria Isoflavone

作　　者：陆瑶 周密 鲁婷婷 郑梅竹[2] LU Yao;ZHOU Mi;LU Ting-ting;ZHENG Mei-zhu(College of Life Sciences,Changchun Normal University,Changchun 130032,China;Central Laboratory,Changchun Normal University,Changchun 130032,China)

机构地区：[1]长春师范大学生命科学学院,吉林长春130032 [2]长春师范大学中心实验室,吉林长春130032

出　　处：《长春师范大学学报》2023年第2期77-83,共7页Journal of Changchun Normal University

基　　金：国家自然科学基金资助项目“药用植物中异黄酮类成分抗缺血性脑卒中的活性筛选及作用机制研究”(31670358);吉林省科技厅基金资助项目“葛花中异黄酮类成分抗缺血性脑卒中的活性筛选及作用机制研究”(20210101031JC);吉林省教育厅基金资助项目“葛花中鸢尾苷抗缺血性脑卒中的活性评价及作用机制研究”(JJKH20220832KJ);长春师范大学研究生科研创新项目“基于网络药理学和分子对接研究射干中野鹫尾苷抗缺血性脑率中的活性评价及作用机制”(〔2022〕第102号)。

摘　　要：利用PC12细胞建立糖氧剥夺(OGD)体外脑卒中损伤模型,采用该模型对葛根异黄酮抗缺血性脑卒中活性进行筛选和验证。评价指标包括:细胞存活率检测、细胞内活性氧(ROS)含量检测、乳酸脱氢酶(LDH)释放率检测、细胞内钙离子(Ca^(2+))浓度检测和细胞凋亡程度检测,并利用蛋白免疫印迹法(Western Blot)检测凋亡蛋白B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)和B淋巴细胞瘤-2基因相关启动子(Bcl-xL/Bcl-2asociated death promoter,BAD)的表达水平。结果显示,与模型组比较,葛根异黄酮保护组能够抑制细胞凋亡,降低细胞内ROS水平,降低LDH释放、抑制细胞内Ca^(2+)累积,并且Bcl-2表达水平升高,BAD表达水平降低,Bcl-2与BAD蛋白表达量比值上调。表明葛根异黄酮能减轻氧糖剥夺/复氧(OGD/R)造成的PC12细胞损伤,提升细胞存活率,葛根异黄酮可能具有一定的抗缺血性脑卒中活性。其作用机制可能与抑制神经细胞的凋亡,减少神经元丢失相关。PC12 cells were used to establish an OGD-induced stroke injury model in vitro,and the anti-ischemic stroke activity of Pueraria Isoflavones was screened and verified by this model.The evaluation indexes include cell survival rate,intracellular reactive oxygen species(ROS)content,lactate dehydrogenase(LDH)release rate,intracellular calcium ion(Ca^(2+))concentration and apoptosis degree.The apoptotic protein B-cell lymphoma-2(Bcl-2)gene and B-cell lymphoma-2 were detected by Western Blot.The results showed that,compared with the model group,pueraria isoflavones protection group and edaravone positive drug control group significantly reduced cell apoptosis,LDH release,intracellular Ca^(2+)accumulation and intracellular ROS level,the level of Bcl-2 increased and the level of BAD decreased,the level of Bcl-2/BAD increased.The results indicated that Pueraria Isoflavones could alleviate the injury of PC12 cells caused by oxygen-glucose deprivation/reoxygenation(OGD/R)and improve the cell survival rate.Pueraria isoflavone may have certain anti-ischemic stroke activity.Its mechanism may be related to the inhibition of neuronal apoptosis and the reduction of neuronal loss.

关 键 词：葛根异黄酮 脑卒中 PC12细胞 

分 类 号：R285.5[医药卫生—中药学]