JNK信号通路对乙醛刺激的大鼠肝星状细胞增殖的影响

Effects of JNK on proliferation of acetaldehyde stimulated hepatic stellate cells in rats

目的: 初步探讨JNK信号通路调控乙醛刺激的大鼠肝星状细胞(HSC)增殖的分子机制.方法: 体外培养大鼠HSC株,用c Jun氨基末端激酶(c- JunN terminalkinase,JNK)特异性阻断剂SP600125对乙醛刺激的大鼠HSC株进行处理,以MTT比色法检测细胞增殖,Western blotting检测磷酸化JNK(p JNK)水平的变化.结果: 乙醛刺激后HSC内p -JNK的水平升高,增殖明显;SP600125对乙醛刺激的HSC对p -JNK的水平有明显下调作用,同乙醛组相比有显著性差异. {MVI比值[ (36. 0±3 .4 ), ( 24. 2±1 .8 ), ( 3. 2±0 .9 ) ]vs( 47. 6±8. 6),P<0. 05},强度呈剂量依赖关系;同时具有抑制细胞增殖的作用: 空白对照组及乙醛组中吸光度(A)为( 0. 07±0 .01)和(0. 16±0 .02),两者有显著性差异(P<0 .01 ),经不同剂量的SP600125 ( 20, 60, 100μg/L)处理后吸光度值下降[分别为(0 .13±0. 01), (0 .12±0. 01), (0 .12±0. 01) ],同乙醛组相比有显著性差异(P< 0 .05 ),细胞抑制率分别为19 .3%, 22. 4%, 28. 6%.结论: JNK活性变化与乙醛刺激的大鼠HSC的增殖呈正相关,抑制JNK活性可影响HSC的增殖,提示JNK信号传导通路在调节HSC的增殖中起着重要的作用.

AIM: To explore the regulating mechanism of c-Jun N-terminal kinase (JNK) signal transduction in the proliferation of hepatic stellate cells stimulated by acetaldehyde.METHODS: Rat hepatic stellate cells (HSCs) stimulated by acetaldehyde were incubated with different concentration of SP600125 in vitro.The cell proliferation was evaluated by MTT colorimetric assay and the variability of JNK was examined by Western Blotting.RESULTS: SP600125 at different concentrations (20 ng/mL,60 ng/mL and 100 ng/mL) suppressed the activity of JNK｛MVI [(36.0±3.4),(24.2±1.8),(3.2±0.9)] vs (47.6±8.6),P<0.05｝,but also inhibited the proliferation of HSCs stimulated by acetaldehyde in a dose-dependent manner [Optical density(A): (0.13±0.01),(0.12±0.01),(0.12±0.01),P<0.05]. CONCLUSION: The proliferation inhibition of HSCs is correlated with the decreased activity of JNK.