丹参通络解毒汤对缺氧/复氧大鼠心肌微血管内皮细胞自噬的影响  

Effects of Danshen Tongluo Jiedu Decoction on Autophagy of Cardiac Microvascular Endothelial Cells in Hypoxia/Reoxygenation Rats

作　　者：王婷婷 李鑫辉 戴超男 李彩云 WANG Tingting;LI Xinhui;DAI Chaonan;LI Caiyun(School of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208

出　　处：《中国中医药信息杂志》2023年第4期65-70,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82074392);湖南省自然科学基金(2019JJ40210)。

摘　　要：目的 观察丹参通络解毒汤对缺氧/复氧大鼠心肌微血管内皮细胞(CMECs)自噬的影响,探讨其保护CMECs的机制。方法 体外培养CMECs,复制缺氧/复氧损伤细胞模型,通过慢病毒转染沉默MALAT1,将细胞分为过表达SRPK1组、过表达SRPK1联合含药血清组、空载组、空载联合含药血清组,Western blot检测自噬相关蛋白Beclin1、LC3B及凋亡相关蛋白Bcl-2、Bax表达,RT-PCR检测LC3B mRNA表达,免疫荧光染色检测LC3B表达。结果 与过表达SRPK1组比较,过表达SRPK1联合含药血清组和空载组细胞Bcl-2蛋白表达显著升高,Bax和Beclin1蛋白表达、LC3BⅡ/LC3BⅠ比值及LC3B mRNA和蛋白表达显著降低(P<0.05,P<0.01);与过表达SRPK1联合含药血清组比较,空载联合含药血清组细胞Bcl-2蛋白表达显著升高,Bax和Beclin1蛋白表达、LC3BⅡ/LC3BⅠ比值及LC3B mRNA和蛋白表达显著降低(P<0.01);与空载组比较,空载联合含药血清组细胞Bcl-2蛋白表达显著升高,Bax和Beclin1蛋白表达、LC3BⅡ/LC3BⅠ比值及LC3B mRNA和蛋白表达显著降低(P<0.01)。结论 丹参通络解毒汤能抑制缺氧/复氧损伤引起的CMECs自噬,进而保护CMECs,其机制可能与下调MALAT1表达,抑制SRPK1表达有关。Objective To observe the effects of Danshen Tongluo Jiedu Decoction on autophagy of cardiac microvascular endothelial cells(CMECs) in hypoxia/reoxygenation rats;To explore the mechanism of its protection of CMECs.Methods CMECs was cultured in vitro to replicate the hypoxia/reoxygenation injury model.MALAT1was silenced by lentiviral transfection,and the cells were divided into overexpression SRPK1 group,overexpression SRPK1 combined with drug-containing serum group,empty-loading group,and empty-loading combined with drugcontaining serum group.Western blot was used to detect the autophagy-related protein Beclin1,LC3B and apoptosisrelated protein Bcl-2,Bax expression;RT-PCR was used to detect the expression of LC3B mRNA;immunofluorescence staining was used to detect the expression of LC3B.Results Compared with the overexpression SRPK1 group,the Bcl-2 protein expression in overexpression SRPK1 combined with the drug-containing serum group and the empty-loading group significantly increased,the protein expressions of Bax and Beclin1,the ratio of LC3BⅡ/LC3BⅠ,and the mRNA and protein expression of LC3B significantly decreased(P<0.05,P<0.01).Compared with the overexpression SRPK1 combined with drug-containing serum group,the expression of Bcl-2 protein in empty-loading combined with drug-containing serum group significantly increased,the protein expressions of Bax and Beclin1,the ratio of LC3BⅡ/LC3BⅠ,and the mRNA and protein expressions of LC3B significantly decreased(P<0.01).Compared with the empty-loading group,the expression of Bcl-2 protein in empty-loading combined with drug-containing serum group significantly increased,the protein expressions of Bax and Beclin1,the ratio of LC3BⅡ/LC3B Ⅰ,and the mRNA and protein expressions of LC3B significantly decreased(P<0.01).Conclusion Danshen Tongluo Jiedu Decoction containing serum can inhibit autophagy of CMECs induced by hypoxia/reoxygenation,then protect CMECs,and its mechanism may be related to down-regulating MALAT1 expression and inhibition SRPK1expression.

关 键 词：丹参通络解毒汤 缺氧/复氧 MALAT1 自噬 SRPK1 心肌微血管内皮细胞 

分 类 号：R285.5[医药卫生—中药学]