δ阿片受体激活通过蛋白激酶C途径抑制人肝细胞凋亡  

δ-opioid receptor activation by protein kinase C pathway inhibits acid-induced liver cell apoptosis

作　　者：唐博[1] 张易[1] 梁锐[1] 隋吉栋[1] 金学利[1] 袁鹏[2] 王立明[1] 

机构地区：[1]大连医科大学附属第二医院,116027 [2]天津医学高等专科学校人体解剖学教研室

出　　处：《中华肝胆外科杂志》2012年第3期211-214,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：国家高技术研究发展计划（863计划）资助（2006AA02A309）

摘　　要：目的研究δ阿片受体激活对人肝细胞凋亡的抑制作用并探讨其与蛋白激酶C（PKC）途径的关系。方法体外培养人肝细胞，用MTT法检测细胞存活率；AnnexinV—FITC／PI双标记法检测细胞凋亡率；流式细胞仪分析细胞周期；RT—PCR检测PKC的mRNA表达；Westernblot分析PKC、Caspase-3蛋白的表达。结果体外培养的人肝细胞经血清剥夺48h后可发生明显的凋亡。激活δ阿片受体可显著抑制这一现象，细胞凋亡率降低（8．37±0．47）、Caspase-3蛋白表达减少（0．147±0．042）、PKC蛋白表达增加（O．698±0．130）。给予PKC拮抗剂GFl09203X后，δ阿片受体抑制细胞凋亡的作用显著下降（20．66±0．60）。结论激活肝细胞膜上的δ阿片受体对血清剥夺导致的人肝细胞凋亡具有抑制作用，其作用机制与PKC通路相关。Objective To study the inhibitory effects of δ-opioid receptor activation in serum deprivation induced apoptosis of human liver cells and the proposed protein kinase C （PKC） pathway mechanism. Methods MTT assay was used to detect the survival rate of human liver cells in vitro and Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate. Flow cytomctry was used to analyze cell cycle, RT PCR used to analyze the PKC mRNA and Western Blot analysis was used for detecting the protein expression of PKC and Caspase-3. Results After serum-deprivation for 48h of cultured human liver cells in vitro, significant liver cell apoptosis occurred. The apoptosis was suppressed by 3-opioid receptor activation, which manifested as a slower rate of apoptosis, decreased expression of Caspase-3and increased expression of PKC. After GF109203X was added, the inhibitory effects of DADLE decreased markedly. Conclusion Activation of δ-opioid receptor on the membrane of human liver cells has inhibitory effects on serum-deprivation induced apoptosis of liver cells. The underlying mechanism may be associated with PKC pathway activation.

关 键 词：阿片受体 肝细胞 凋亡 CASPASE-3 蛋白激酶C 

分 类 号：R285.5[医药卫生—中药学]