摘要
目的探究丹酚酸B(salvianolic acid B,Sal B)减轻缺氧/复氧(H/R)诱导的大鼠肝细胞损伤及潜在的分子机制。方法体外培养BRL-3A大鼠肝细胞株,制备BRL-3A的H/R模型,予以Sal B干预。CCK-8检测细胞活力;微板法测转氨酶含量;ELISA测炎性因子的含量;流式细胞术测细胞凋亡水平;Western blot和实时荧光定量PCR(q PCR)检测SIRT1、NF-κB p65、p53、Bax、Bcl-2蛋白及mRNA表达水平。结果 H/R诱导的大鼠肝细胞活力下降;细胞上清液中ALT、AST、TNF-α、IL-1β含量明显增多;细胞凋亡率明显升高;SIRT1和Bcl-2在蛋白及mRNA水平的表达下调,而NF-κB p65、p53和Bax在蛋白及mRNA水平的表达上调。在Sal B预处理后,细胞活力升高;细胞液中ALT、AST、TNF-α及IL-β的含量下降;细胞凋亡率降低;SIRT1和Bcl-2在蛋白水平及mRNA水平的表达升高,NF-κB p65、p53和Bax在蛋白水平及mRNA水平的表达降低。结论丹酚酸B可有效减轻H/R诱导的大鼠肝细胞损伤,其机制可能与SIRT1/NF-κB/p53通路有关系。
Aim To investigate the effect of salvianolic acid B(Sal B)on the attenuation of rat hepatocyte injury induced by hypoxia/reoxygenation(H/R)and its possible molecular mechanism.Methods Rat hepatocytes BRL-3A were cultured in vitro.H/R injury model was established and then BRL-3A cells were pretreated with Sal B.The viability of cells was measured by CCK-8 assay;the expression of ALT and AST was detected by microplate assay;the levels of TNF-αand IL-1βwere determined by ELISA;the apoptosis was detected by flow cytometry;the protein and mRNA levels of SIRT1,NF-κB p65,p53,Bax and Bcl-2 were measured by Western blot and qPCR.Results H/R intervention decreased the viability and increased the apoptosis of cells;the production of ALT,AST,TNF-αand IL-1βwas elevated;the protein and mRNA levels of SIRT1,Bcl-2 were reduced,but the levels of NF-κB p65,p53 and Bax increased.After pretreated with Sal B,the viability of cells increased while the apoptosis decreased;the expression of ALT,AST,TNF-αand IL-1βwas inhibited;moreover,the protein and mRNA levels of SIRT1,Bcl-2 were enhanced,and the levels of NF-κB p65,p53 and Bax decreased significantly.Conclusion Sal B may attenuate rat hepatocyte injury induced by H/R via the SIRT1/NF-κB/p53 pathway.
作者
万磊
陈青松
周壮
周翔宇
郑道峰
吴忠均
WAN Lei;CHEN Qing-song;ZHOU Zhuang;ZHOU Xiang-yu;ZHENG Dao-feng;WU Zhong-jun(Dept of Hepatobiliary Surgery,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第5期680-685,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81672959)
重庆市科学技术委员会基金项目(No cstc2015shmszx120019)
