羟基红花红色素A抑制氧化应激和细胞凋亡改善OGD/R诱导的HT22细胞损伤  

Hydroxysafflor yellow A protects against OGD/R-induced HT22 cell injury by suppressing oxidative stress and apoptosis

作　　者：袁美玲 张云 王艳 汪宁 YUAN Mei-ling;ZHANG Yun;WANG Yan;WANG Ning(Anhui University of Chinese Medicine,Hefei 230012;Anhui Key Laboratory of Research&Development of Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230012;Institute for Pharmacodynamics and Safety Evaluation of Chinese Medicine,Anhui Academy of Traditional Chinese Medicine,Hefei 230012)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]安徽省中药研究与开发重点实验室,合肥230012 [3]安徽省中医科学院中药药效与安全性评价研究所,合肥230012

出　　处：《中南药学》2022年第1期1-7,共7页Central South Pharmacy

基　　金：国家自然科学基金面上项目(No.81773933);安徽省高校自然科学研究重点项目(No.KJ2020A0410);安徽省高校自然科学研究重点项目(No.KJ2019A0434);安徽省第十三批115产业创新团队“针药结合防治脑病创新团队”项目(皖人才办[2020]4号)。

摘　　要：目的探讨羟基红花红色素A(HSYA)对氧糖剥夺/复糖复氧(OGD/R)损伤小鼠海马神经元(HT22)细胞的保护作用及其作用机制。方法体外培养HT22细胞,随机分为5组,分别为正常组,OGD/R组,HSYA低、中、高给药组(浓度分别为40、60、80μmol·L^(-1))。除正常组外,其余各组细胞在氧糖剥夺6 h后迅速复糖复氧12 h进行OGD/R造模并分组给药。采用CCK8法检测HT22细胞活力,倒置显微镜观察HT22细胞形态。试剂盒检测LDH、NO、GSH、MDA、SOD水平。采用Hoechst染色和流式细胞术检测HT22细胞凋亡情况,ELISA检测CytC释放情况,Western blot检测Bax、Bcl-2和cleaved-caspase3的蛋白表达情况,qRTPCR检测Bax和Bcl-2的mRNA水平。结果与OGD/R组相比,HSYA低、中、高给药组细胞存活率显著升高(P<0.05,P<0.01),细胞形态显著改变,HT22细胞LDH、NO、MDA及细胞凋亡率显著降低,SOD和GSH水平升高(P<0.05,P<0.01),胞浆内CytC释放量显著降低(P<0.01),Bax的蛋白表达和mRNA水平显著降低,Bcl-2的蛋白表达和mRNA水平显著升高,cleaved-caspase3的蛋白表达降低(P<0.05,P<0.01)。结论HSYA通过抑制氧化应激和细胞凋亡改善OGD/R诱导的HT22细胞损伤。Objective To determine the protective effect of hydroxysafflor yellow A(HSYA)on hippocampal neuron(HT22)cells injured by OGD/R and related mechanism.Methods HT22 cells were cultured in vitro and randomly divided into 5 groups:a normal group,an OGD/R group,and HSYA low,medium,and high dose groups(40,60,and 80μmol·L^(-1)).Except for the normal group,the cells in the other groups were quickly reoxygenated for 12 hours after oxygen and glucose deprivation for 6 hours.The OGD/R model was established and the drugs were given in groups.The viability of HT22 cells was detected by CCK8 method,and the morphology of HT22 cells was observed by inverted microscope.The levels of NO,GSH,MDA,LDH and SOD in the cells were detected by test kits.The apoptosis of HT22 cells was detected by Hoechst staining and flow cytometry,the release of Cytc was detected by ELISA;the protein expression of Bax,Bcl-2 and cleaved-caspase3 was detected by Western blot,and the mRNA levels of Bax and Bcl-2 were detected by qRT-PCR.Results Compared with the OGD/R group,the cell survival rates of the HSYA groups were significantly increased(P<0.05,P<0.01),and the cell morphology was significantly changed.LDH,NO,MDA and the apoptosis rates of HT22 cells were significantly decreased.The levels of SOD and GSH were significantly increased(P<0.05,P<0.01),and the Cytc release was significantly decreased(P<0.05,P<0.01).The protein expression and mRNA level of Bax were significantly decreased,while those of Bcl-2 increased.The protein expression of cleaved-cspase3 was decreased(P<0.05,P<0.01).Conclusion HSYA ameliorates OGD/R-induced HT22 cell injury by inhibiting oxidative stress and apoptosis.

关 键 词：羟基红花红色素A 氧糖剥夺/复糖复氧 HT22细胞 氧化应激 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]