PI3K/Akt信号传导通路在乙醛刺激下大鼠肝星状细胞株HSC-T6增殖中的作用

Effects of PI3K/Akt signaling pathway on acetaldehyde-stimulated proliferation of hepatic stellate cells HSC-T6

目的探讨磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号传导通路在乙醛刺激的大鼠肝星状细胞株HSC-T6增殖中的作用。方法将大鼠HSC-T6随机分为6组:空白对照组(A组):采用10%胎牛血清的DMEM培养液培养;乙醛组(B组):在A组基础上加乙醛,使终浓度为200μmol/L;实验组在B组基础上加入不同浓度(25、50、75、100μmol/L)的PI3K/Akt通路抑制剂LY294002,分别称为C、D、E、F组。用CCK-8法检测大鼠HSC-T6增殖活力(用A值表示)。再随机将HSC-T6分为3组:空白对照组(10%胎牛血清的DMEM培养液培养)、乙醛组(在空白对照组基础上加乙醛,使终浓度为200μmol/L)、乙醛+LY294002组(在乙醛组的基础上加入50μmol/L LY294002),用Western blotting法检测大鼠HSC-T6中PI3K、p-Akt蛋白的表达情况。结果 A、B、C、D、E、F组A值分别为0.545 0±0.024 4、0.952 3±0.045 5、0.927 3±0.041 9、0.865 8±0.028 9、0.814 3±0.024 7、0.759 3±0.220 0。与A组比较,其他各组A值均升高;与B、C组比较,D、E、F组降低(P均<0.01)。与空白对照组比较,乙醛组、乙醛+LY294002组PI3K、p-AKT蛋白相对表达量高(P均<0.01),与乙醛组比较,乙醛+LY294002组降低(P<0.05)。结论 PI3K/Akt信号传导通路部分参与调控乙醛刺激的大鼠HSC-T6增殖。

Objective To investigate the effects of phosphatidylinositol 3-kinase（ PI3K）/Akt signaling pathway specific inhibitor LY294002 on the proliferation of rat hepatic stellate cells（ HSC-T6） stimulated by acetaldehyde. Methods The rat HSC-T6 cells were randomly divided into six groups： the blank control group（ group A） ： we used DMEM culture solution with 10% fetal bovine serum（ FBS） to culture,the acetaldehyde group（ group B） ： we added acetaldehyde on the basis of the blank control group and made the final concentration be 200 μmol/L,the experimental group was added with different concentrations of PI3 K/Akt pathway specific inhibitor LY294002（ 25,50,75 and 100 μmol/L） on the basis of acetaldehyde group（ group B）,and then they were named groups C,D,E and F,respectively. CCK-8 was applied to detect the proliferation activity of rat HSC-T6 cells. And then HSC-T6 cells were randomly divided into three groups： the blank control group（ added with DMEM culture solution with 10% fetal bovine serum）,acetaldehyde group（ added with acetaldehyde on the basis of blank control group and made the final concentration be 200 μmol/L）,and acetaldehyde ＋LY294002 group（ added with 50 μmol/L PI3 K/Akt pathway specific inhibitor LY294002 on the basis of acetaldehyde group）. Western blotting was used to detect the expression of PI3 K and p-Akt protein in rat HSC-T6. Results The A values of groups A,B,C,D,E,and F were 0. 545 0 ± 0. 024 4,0. 952 3 ± 0. 045 5,0. 927 3 ± 0. 041 9,0. 865 8 ± 0. 0289,0. 814 3 ± 0. 024 7 and 0. 759 3 ± 0. 220 0,respectively. Compared with group A,the A values of other groups all increased; compared with groups B and C,the A values of groups D,E and F all decreased（ all P〈0. 01）. Compared with the blank control group,the protein expression of PI3 K and p-Akt in acetaldehyde group and acetaldehyde ＋ LY294002 group were increased（ all P〈0. 01）. Compared with the acetaldehyde group,the value of acetaldehyde ＋ LY294002 group decreased（ P〈0. 05）. Conclusion PI3 K/Akt signaling pathway may partly adjust and control the proliferation of rat HSC-T6 stimulated by acetaldehyde.