【摘要】 目的:探讨内质网应激对肝星状细胞分泌肝细胞生长因子(hypatocyte growth factor,HGF)的影响及其可能机制。方法 :通过在培养的大鼠肝星状细胞T6中分别加入5.0μg/m L衣霉素或0.2μmol/L毒胡萝卜素建立肝星状细胞内质网应激模型,4.0 mmol/L 4-苯基丁酸钠(4-phenylbutyrate,4-PBA)和200.0μmol/L salubrinal作为内质网应激抑制剂对T6细胞进行预处理,重组慢病毒LV-e If2α-sh RNA-GFP敲减T6细胞中的e If2αm RNA表达,并提取m RNA和全蛋白进行RT-PCR和Western blot实验检测HGF、分子伴侣重链结合蛋白(glucose-regulated protein 78,GRP78)、真核翻译起始因子2α(eukaryotic translation initiation factor 2α,e If2α)、磷酸化e If2α、激活转录因子4(activating transcription factor 4,ATF4)以及凋亡信号分子C/EBP同源蛋白(C/EBP homologous protein,CHOP)。结果:衣霉素、毒胡萝卜素可诱导T6细胞GRP78升高,激活内质网应激状态的同时抑制HGF表达,4-PBA和salubrinal可阻止内质网应激引起的HGF降低,但对ATF4和CHOP的表达作用不同。慢病毒转染T6降低细胞e If2α表达的同时成比例降低HGF表达。结论:ERS激活后可通过影响e If2α表达从而抑制HGF表达。更多还原

【Abstract】 Objective: To detect whether suppression of endoplasmic reticulum(ER) stress maintains hepatocyte growth factor(HGF) expression in hepatic stellate cells(HSCs) and its potential mechanism. Methods: Rat hepatic stellate cell line HSC-T6 was treated with the ER stress agonists 5.0 μg / m L tunicamycin and 0.2 μmol / L thapsigargin, and ER stress inhibitors sodium 4-phenylbutyrate(4-PBA) 4.0 mmol / L and salubrinal 200.0 μmol / L were used as pretreatment. Recombinant lentivirus LV-e If2α-sh RNAGFP was produced to block e If2α activated by ER stress. Levels of HGF, glucose-regulated protein 78(GRP78), eukaryotic translation initiation factor 2α(e If2α), phospho-e IF2α, activating transcription factor 4(ATF4) and C / EBP(CCAAT / enhancer binding protein)homologous protein(CHOP) in vitro were measured by quantitative RT-PCR and Western blot. Results: Our results demonstrated that tunicamycin or thapsigargin stimulated GRP78 expression, and activation of ER stress inhibited HGF expression in HSC-T6 cells. The inhibition of HGF could be partly prevented in the presence of 4-PBA or salubrinal, but their effects on ATF4 and CHOP expression were different. Interfering e If2α m RNA proportionately down-regulated HGF expression. Conclusion: The activation of ER stress inhibits HGF expression of HSCs through decreasing e IF2α expression.更多还原