论文部分内容阅读
目的:探讨缺氧在肝星状细胞(HSC)激活中的作用机制。方法:缺氧培养大鼠HSC株HSC-T6,逆转录PCR检测HIF-1α、TGF-β1、I型胶原mRNA的表达,Westren blot检测α-SMA表达。缺氧/常氧下,加TGF-β1中和抗体、HIF-1α诱导剂氯化钴(CoCl2)或抑制剂2-甲氧雌二醇(2ME2),逆转录PCR检测HSC-T6表达I型胶原mRNA的变化。结果:缺氧能诱导HSC-T6表达HIF-1α、TGF-β1、I型胶原mRNA及α-SMA蛋白;常氧下CoCl2能诱导HSC-T6表达I型胶原mR-NA,TGF-β1中和抗体及2ME2能减弱缺氧诱导HSC-T6表达I型胶原mRNA,但表达量仍高于常氧培养。结论:缺氧能激活HSC,分泌I型胶原,增加细胞外基质沉积,其作用机制既依赖于TGF-β的胞内信号传导机制,也与HIF-α介导的信号传导有关,在肝纤维化形成中起着举足轻重的作用。
Objective: To explore the mechanism of hypoxia in the activation of hepatic stellate cells (HSC). Methods: HSC-T6 was cultured in hypoxia, the expression of HIF-1α, TGF-β1 and collagen I was detected by RT-PCR and the expression of α-SMA was detected by Westren blot. Under hypoxia / normoxia, TGF-β1 neutralizing antibody, HIF-1αinducing agent cobalt chloride (CoCl2) or 2ME2 inhibitor 2ME2 were detected. The expression of HSC-T6 was detected by reverse transcription polymerase chain reaction Collagen mRNA changes. RESULTS: Hypoxia could induce HSC-T6 to express HIF-1α, TGF-β1, type I collagen mRNA and α-SMA protein. CoCl2 could induce mR-NA, TGF- Antibody and 2ME2 attenuated hypoxia-induced HSC-T6 expression of type I collagen mRNA, but the expression level was still higher than that of normoxic culture. Conclusion: Hypoxia activates HSC, secretes type I collagen and increases extracellular matrix deposition. The mechanism of action is dependent on the intracellular signal transduction mechanism of TGF-β and on the signal transduction mediated by HIF-α. The formation of play a decisive role.