PAX6通过MEK/ERK信号通路抑制肝星状细胞活化和增殖实验研究  

PAX6 inhibiting activation and proliferation of hepatic stellate cells through MEK/ERK signaling pathway in vitro

作　　者：白亮[1] 王宝太[1] 高志峰 蒋安[1] 梁金强 Bai Liang;Wang Baotai;Gao Zhifeng(Department of General Surgery,Second Affiliated Hospital,Jiaotong University,Xi'an 710000,Shaanxi Province,China)

机构地区：[1]西安交通大学第二附属医院普外科,西安市710000

出　　处：《实用肝脏病杂志》2022年第3期318-322,共5页Journal of Practical Hepatology

摘　　要：目的探讨PAX6通过丝裂原活化蛋白激酶(MEK)/细胞外调节蛋白激酶(ERK)信号通路抑制肝星状细胞活化和增殖的效果。方法取LX2肝星状细胞,分为对照组、PAX6 inhibitor组和PAX6 mimics组。采用CCK-8试剂盒测定细胞增殖,采用油红O染色测定细胞分化水平,使用流式细胞仪分析细胞凋亡,采用RT-PCR法和蛋白印迹法测定细胞PAX6、MEK和ERK mRNA和蛋白表达水平。结果PAX6 inhibitor组细胞PAX6 mRNA水平、凋亡率和G1期分别为(1.49±0.23)、(2.70±0.85)%和(59.02±1.25)%,显著低于LX2肝星状细胞组【分别为(1.85±0.19)、(3.40±0.47)%和(64.66±1.41)%,P<0.05】,而细胞增殖、存活率、分化率、MEK和ERK mRNA和蛋白水平分别为(0.79±0.03)、(73.35±9.74)%、(49.37±4.24)%、(2.55±0.43)、(3.90±0.49)、(0.89±0.15)和(1.17±0.17),显著高于LX2肝星状细胞组【分别为(0.58±0.05)、(60.74±9.24)%、(29.35±4.47)%、(2.67±0.47)、(4.55±0.50)、(0.74±0.14)和(1.35±0.16),P<0.05】;PAX6 mimics组细胞PAX6 mRNA水平、凋亡率和G1期分别为(2.67±0.20)、(6.70±1.04)%和(66.38±1.35)%,显著高于LX2肝星状细胞组【分别为(1.85±0.19)、(3.40±0.47)%和(64.66±1.41)%,P<0.05】,而细胞增殖、存活率、分化率、MEK和ERK mRNA和蛋白水平分别为(0.40±0.04)、(52.24±5.57)%、(13.85±3.35)%、(1.94±0.53)、(1.45±0.42)、(0.53±0.15)和(0.53±0.16),显著低于LX2肝星状细胞组【分别为(0.58±0.05)、(60.74±9.24)%、(29.35±4.47)%、(2.67±0.47)、(4.55±0.50)、(0.74±0.14)和(1.35±0.16),P<0.05】。结论PAX6过表达可抑制肝星状细胞活化和增殖,促进其凋亡,其机制可能与PAX6过表达可抑制肝星状细胞MEK和ERK表达进而抑制了MEK/ERK通路的激活有关。Objective The purpose of this experiment was to investigate the effect of PAX6 on inhibition of activation and proliferation of hepatic stellate cells(HSCs)by through MEK/ERK signaling pathway.Methods The LX2 HSCs were divided into control,PAX6 inhibitor and PAX6 mimics group and managed accordingly.The cell proliferation was determined by CCK-8 kit,the cell differentiation was detected by oil red O staining,the cell apoptosis was analyzed by flow cytometry,and PAX6,MEK and ERK mRNA and their protein expression were assayed by RT-PCR and Western blotting,respectively.Results The PAX6 mRNA level,apoptosis rate and G1 stage in PAX6 inhibitor-intervened group were(1.49±0.23),(2.70±0.85)%and(59.02±1.25)%,significantly lower than[(1.85±0.19),(3.40±0.47)% and(64.66±1.41)%,respectively,P<0.05]in LX2 HSCs,and the cell proliferation,survival rate,differentiation rate,MEK and ERK mRNA and their protein expression in PAX6 inhibitor-intervened group were(0.79±0.03),(73.35±9.74)%,(49.37±4.24)%,(2.55±0.43),(3.90±0.49),(0.89±0.15)and(1.17±0.17),significantly higher than[(0.58±0.05),(60.74±9.24)%,(29.35±4.47)%,(2.67±0.47),(4.55±0.50),(0.74±0.14)and(1.35±0.16),respectively,P<0.05]in the control;the PAX6 mRNA level,apoptosis rate and G1 stages in PAX6 mimics-intervened group were(2.67±0.20),(6.70±1.04)% and(66.38±1.35)%,significantly higher than[(1.85±0.19),(3.40±0.47)%and(64.66±1.41)%,respectively,P<0.05]in the control,and the cell proliferation,survival rate,differentiation rate,MEK and ERK mRNA and their protein expression in PAX6 mimics group were(0.40±0.04),(52.24±5.57)%,(13.85±3.35)%,(1.94±0.53),(1.45±0.42),(0.53±0.15)and(0.53±0.16),significantly lower than[(0.58±0.05),(60.74±9.24)%,(29.35±4.47)%,(2.67±0.47),(4.55±0.50),(0.74±0.14)and(1.35±0.16),respectively,P<0.05]in the control.Conclusion The PAX6 overexpression could inhibit the activation and proliferation of hepatic stellate cells,and promote cell apoptosis.The mechanism involved might be related to the inhibition of MEK and ERK e

关 键 词：肝星状细胞 PAX6 MEK/ERK信号通路 细胞活化 细胞增殖 体外 

分 类 号：R575.2[医药卫生—消化系统]