过表达Perilipin5和抑制PKA对大鼠肝星状细胞凋亡的影响  

Effects of PKA on apoptosis of rat hepatic stellate cells induced by Perilipin5

作　　者：殷雪翠 汤有才[2,3,4,5] 李哲[6] 李恩耀[7] 李元枭 李璇璇 郑鹏远[1] YIN Xuecui;TANG Youcai;LI Zhe;LI Enyao;LI Yuanxiao;LI Xuanxuan;ZHENG Pengyuan(Department of Gastroenterology,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Pediatrics,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Children′s Rehabilitation Medicine,the Fifth Affiliated Hospital,ZhengzhouUniversity,Zhengzhou 450052;Henan Key Laboratory of Rehabilitation Medicine,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Henan Joint International Research Laboratory of Chronic Liver Injury,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Rehabilitation,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Children with Cerebral Palsy,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第五附属医院消化内科,郑州450052 [2]郑州大学第五附属医院儿科,郑州450052 [3]郑州大学第五附属医院儿童康复科,郑州450052 [4]郑州大学第五附属医院河南省康复医学重点实验室,郑州450052 [5]郑州大学第五附属医院河南省慢性肝损伤国际联合实验室,郑州450052 [6]郑州大学第五附属医院康复科,郑州450052 [7]郑州大学第五附属医院儿童脑瘫科,郑州450052

出　　处：《郑州大学学报（医学版）》2020年第2期207-212,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金面上项目(31471330);河南省科技厅国际合作项目(172102410014);河南省教育厅重点科研项目(18A320011);郑州市协同创新重大专项(18XTZX12003)。

摘　　要：目的:探讨蛋白激酶A(PKA)信号通路是否参与Perilipin5(Plin5)抑制肝星状细胞(HSC)增殖和诱导其凋亡。方法:将大鼠原代HSC(HSC-Primary)和细胞株HSC-T6感染Plin5慢病毒,Western blot法检测细胞中Plin5、t-PKA、磷酸化PKA(p-PKA)的表达。将两种细胞分别分成4组:空白对照组(不感染)、过表达Plin5组(感染Plin5慢病毒)、H89组(用10 mg/L的PKA抑制剂H89处理),Plin5+H89共处理组(感染Pin5慢病毒后用H89处理)。分组处理48 h后,采用Western blot法检测细胞中Cyclin D1、P21、Bax和Bcl-2表达水平,MTT法检测细胞增殖率,检测细胞中Caspase-3活性。结果:两种细胞感染Plin5慢病毒后Plin5表达水平和PKA磷酸化水平均较未感染细胞增强(P<0.05)。与空白对照组比较,过表达Plin5组细胞增殖率降低,P21和Bax蛋白表达上调,Cyclin D1和Bcl-2蛋白表达下调,Caspase-3活性升高(P<0.05);上述变化均可被H89阻断(P<0.05)。结论:Plin5通过激活PKA通路,抑制HSC增殖并诱导其凋亡;Plin5可能成为抑制肝纤维化的作用靶点。Aim:To explore the role of Perilipin5(Plin5)on the proliferation inhibition and apoptosis induction in hepatic stellate cells(HSC)and its relation with PKA signaling pathway.Methods:Rat primary HSC(HSC-Primary)and cell line HSC-T6 were infected with Plin5 lentivirus,and Western blot was used to detect the expressions of Plin5,PKA(t-PKA),and phosphorylated PKA(p-PKA).HSC-Primary and HSC-T6 cells were allocated into 4 groups,respectively:blank control group(without infection),Plin5 overexpression group(infected with Plin5 lentivirus),H89 group(treated with 10 mg/L PKA inhibitor H89),Plin5+H89 co-processing group(treated with H89 after infection with Plin5 lentivirus).After grouping for 48 hours,the expressions of Cyclin D1,P21,Bax,and Bcl-2 proteins in the cells were detected by Western blot,the cell proliferation rate was detected by MTT method,and the Caspase-3 activity was detected by the Caspase-3 kit.Results:Plin5 expression and PKA phosphorylation level of the cells infected with Plin5 lentivirus were higher than those of uninfected cells(P<0.05).Compared with the blank control group,the cell proliferation rate of the Plin5 overexpression group was reduced,the expressions of P21 and Bax increased,the expressions of Cyclin D1 and Bcl-2 were down-regulated,and Caspase-3 activity increased(P<0.05);all of the above changes could be blocked by H89(P<0.05).Conclusion:Plin5 can inhibit the proliferation and induce apoptosis of HSC by activating the PKA signaling pathway;Plin5 may become a target for the inhibition of liver fibrosis.

关 键 词：肝星状细胞 肝纤维化 信号传导 Perilipin5 PKA 大鼠 

分 类 号：R575[医药卫生—消化系统]