二氢姜黄素体外处理BRL-3A细胞对细胞凋亡通路的影响  

Effects of dihyd rocurcumin on apoptotic pathway in palmitic acid-induced BRL-3A cell apoptosis in vitro

作　　者：吴刚 陈训军 张万里[2] 李芬[2] Wu Gang;Chen Xunjun;Zhang Wanli(Department of Pharmacy,People’s Hospital,Jingshan 431899,Hubei Province,China)

机构地区：[1]湖北省京山市人民医院药剂科,431899 [2]华中科技大学同济医学院附属协和医院肿瘤中心

出　　处：《实用肝脏病杂志》2020年第5期622-625,共4页Journal of Practical Hepatology

基　　金：湖北省自然科学基金资助项目(编号:WJ2015MB184)。

摘　　要：目的研究二氢姜黄素(DHC)对棕榈酸(PA)诱导的大鼠肝BRL-3A细胞线粒体凋亡通路的影响。方法建立PA诱导的非酒精性脂肪性肝炎(NASH)细胞模型,将细胞分为对照组(正常培养基)、模型组(0.25 mM PA处理)和DHC处理组(16、32和64μM DHC处理),检测细胞培养上清乳酸脱氢酶(LDH)含量,采用流式细胞仪检测细胞凋亡,检测细胞线粒体膜电位、凋亡相关蛋白酶Caspase-3和Caspase-9活性,采用Western blotting法检测线粒体凋亡相关蛋白Bcl-2和Bax蛋白表达量。结果模型组细胞上清LDH、Caspase-3和Caspase-9活性分别为(100.1±4.2)%、(1.0±0.2)%和(1.0±0.1)%,均显著高于对照组[分别为(65.2±3.5)%、(0.3±0.1)%和(0.5±0.1)%,P<0.05],而线粒体膜电位为(0.5±0.1)%,显著低于对照组[(0.7±0.1)%,P<0.05];32μM DHC处理组LDH、Caspase-3和Caspase-9活性分别为(84.4±2.4)%、(0.8±0.1)%和(0.7±0.1)%,均显著低于模型组(P<0.05),而线粒体膜电位为[(0.6±0.1)%,显著高于模型组(P<0.05);与对照组比,模型组细胞凋亡率显著增加,而经DHC处理组,细胞凋亡率比模型组显著降低;模型组Bcl-2蛋白相对表达量为(1.0±0.1),显著低于对照组[(2.2±0.5),P<0.05],而Bax蛋白相对表达量为(1.1±0.1),显著高于对照组[(0.8±0.1),P<0.05];32μM DHC处理组和64μM DHC处理组Bcl-2蛋白相对表达量分别为(1.3±0.1)和(1.9±0.2),均显著高于模型组(P<0.05),而Bax蛋白相对表达量分别为(0.7±0.1)和(0.6±0.1),均显著低于模型组(P<0.05)。结论DHC可能通过抑制线粒体介导的凋亡通路,缓解由PA诱导的BRL-3A细胞凋亡。Objective The aim of this study was to investigate the effects of dihyd rocurcumin(DHC)on apoptotic pathway in palmitic acid(PA)-induced BRL-3A cell apoptosis in vitro.Methods The non-alcoholic steatohepatitis(NASH)cell model was established by PA induction in BRL-3A cells.The survival rate of BRL-3A cells was detected by MTT assay.The cells were divided into control with normal medium,model(0.25 mM PA)and DHC intervention group with 16,32,64μM DHC culture.The content of lactate dehydrogenase(LDH)in cell culture supernatant was detected.The apoptosis was detected by flow cytometry.The cell mitochondrial membrane potential(MMP),activity of apoptosis-related protease Caspase-3 and Caspase-9 were detected and the protein expression of mitochondrial apoptosis-related proteins Bcl-2 and Bax was detected by Western blotting.Results The supernatant LDH level,activities of Caspase-3 and Caspase-9 in model group were(100.1±4.2)%,(1.0±0.2)%and(1.0±0.1)%,much higher than[(65.2±3.5)%,(0.3±0.1)%and(0.5±0.1)%,P<0.05]in the control,while the MMP was lower than that in control group[(0.5±0.1)%vs.(0.7±0.1)%,P<0.05];the LDH,Caspase-3 and Caspase-9 in 32μM DH-intervened cells were(84.4±2.4)%,(0.8±0.1)%and(0.7±0.1)%,significantly lower than those in the model(P<0.05),whilethe MMP was(0.6±0.1)%,much higher than that in the model(P<0.05);the apoptotic rate in the model group was significantly increased as compared to that in the control,while it decreased greatly as compared with model in DHC-intervened group;the relative expression of Bcl-2 protein in model group was lower than that in control group[(1.0±0.1)vs.(2.2±0.5),P<0.05],while the relative expression of Bax protein was higher than that in control group[(1.1±0.1)vs.(0.8±0.1),P<0.05],but the relative expression of Bcl-2 protein in 32μM DHC-treated group and 64μM DHC-treated group were(1.3±0.1)and(1.9±0.2),much higher than those in the model group(P<0.05),and the relative expression of Bax protein were(0.7±0.1)and(0.6±0.1),much lower than those in the mod

关 键 词：BRL-3A细胞 二氢姜黄素 非酒精性脂肪性肝炎 线粒体 凋亡通路 

分 类 号：R285.5[医药卫生—中药学]