咖啡因对乙醛诱导的HSC-T6中TGF-β1,CTGF信号转导通路的影响

目的探讨咖啡因(caffeine)对乙醛诱导的大鼠肝星状细胞系(Hepatic Stellate Cell-T6,HSC-T6)中转化生长因子-β1(Trans-forming Growth Factor-β1,TGF-β1),结缔组织生长因子(Connective Tissue Growth Factor,CTGF)信号转导通路的影响。方法实验设正常组(常规培养),模型组及腺苷受体(Adenosine Receptor,AR)调节剂干预组。分别给予caffeine(4 mmol.L-1)1-2],腺苷A2A受体拮抗剂ZM241385(1μmol.L-1)3],腺苷A2A受体激动剂CGS21680(1μmol.L-1)3],caffeine+CGS21680,ZM241385+CGS21680与HSC-T6共同培养,1 h后加入终浓度200μmol.L-1的乙醛刺激(每12 h补充1次),继续培养48 h。采用免疫细胞化学法检测HSC-T6中α-平滑肌肌动蛋白(α-SMA)的表达,RT-PCR法检测TGF-β1和CTGF mRNA水平,Western blot方法检测各组HSC-T6中CTGF蛋白表达。结果与模型组比较,caffeine及ZM241385均显著降低HSC-T6中α-SMA,TGF-β1和CTGF的表达,而caffeine及ZM241385合用CGS21680上述作用有所逆转。结论 Caffeine能够显著降低乙醛诱导的HSC-T6活化,并且显著抑制HSC-T6中TGF-β1和CTGF的表达水平,其机制可能与拮抗腺苷A2A受体介导的信号通路有关。

Effect of caffeine on signaling transduction of TGF-β1 and CTGF in hepatic stellate cell-T6 stimulated by acetaldehyde

Objective To explore the effect of caffeine on proliferation in hepatic stellate cell stimulated by acetaldehyde and its signaling pathway of TGF-β1 and CTGF.Methods Trials are divided into normal group(regular training),model group,adenosine receptor group,which were given caffeine(4 mmol·L-1),adenosine A2A receptor antagonist ZM241385(1 μmol·L-1),adenosine A2A receptor agonists CGS21680(1 μmol·L-1),caffeine+CGS21680,ZM241385+CGS21680 and HSC-T6 respectively,stimulated by acetaldehyde after 1 h,before continueing to cultivate 48 h.The protein expression of α-SMA was analyzed by immunocytochemistry methods.The mRNA expressions of TGF-β1 and CTGF were measured by RT-PCR.The protein expression of CTGF was analyzed by western blot methods.Results Caffeine or ZM241385 treatment inhibited the increase expressions of TGF-β1,CTGF,α-SMA in the HSC-T6,likewise,as with CGS21680 plus caffeine or ZM241385 groups,caffeine or ZM241385 prevented the adenosine A2A receptor agonist from stimulating an increase in hepatic stellate cell.Conclusion Caffeine can suppress the activation of α-SMA in HSC-T6 stimulated by acetaldehyde.Caffeine treatment inhibited the increase expressions of TGF-β1 and CTGF in the HSC-T6,the mechanisms of which may be related to the expression of adenosine A2A receptor.