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H2S调节miR-21表达抑制内质网应激介导肺成纤维化细胞凋亡 被引量:6

Hydrogen sulfide inhibits endoplasmic reticulum stress-mediated apoptosis of pulmonary fibroblasts by regulating expression of microRNA-21
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摘要 目的:探讨H_2S是否可调节mi R-21表达抑制内质网应激介导肺成纤维细胞凋亡。方法:体外培养小鼠成纤维细胞(NIH3T3),随机分为对照组、TGF-β1组和Na HS干预组。采用CCK-8分别检测细胞增殖,流式细胞仪检测NIH3T3凋亡率; q RT-PCR法和Western blot分别分析葡萄糖调节蛋白78(GRP78)、增强子结合蛋白同源蛋白(CHOP)和caspase-12的表达;检测mi R-21是否参与H_2S抑制ERS介导的肺成纤维化细胞凋亡,将其随机分为3组:对照组、mi R-21激动剂(mi R-21 agomir)组和mi R-21拮抗剂(mi R-21 antagomir)组。mi R-21激动剂组和mi R-21拮抗剂组分别给予40μmol/L的Na HS预处理,再进行TGF-β1处理,检测GRP78、CHOP及caspase-12的表达。结果:与对照组比较,Na HS干预组小鼠肺成纤维化细胞的增殖率、细胞凋亡率、mi R-21蛋白及m RNA的表达均明显降低;与TGF-β1组比较,Na HS可明显降低内质网应激相关因子GRP78、CHOP及caspase-12蛋白和m RNA表达;与阴性对照组比较,mi R-21 agomir组GRP78、CHOP及caspase-12的蛋白及m RNA表达均显著升高。而与mi R-21 agomir组比较,mi R-21 antagomir组结果则与之相反。结论:H_2S可通过调节mi R-21的表达,调控TGF-β1诱导小鼠肺成纤维化细胞的ERS稳态减少细胞凋亡,发挥其内源性的保护作用。 Objective:To explore whether H 2S regulating the expression of miR-21 and then inhibition endoplasmic reticulum stress-mediated apoptosis of pulmonary fibroblasts.Methods:Mice pulmonary fibroblasts NIH3T3 were cultured in vitro.Then the cells were divided into a control group,a TGF-β1 group,and an H 2S protection group.The rate of cell proliferation was monitored by CCK-8,and the apoptosis rate of the pulmonary fibroblasts by flow cytometry.The mRNA and protein expressions of GRP78,CHOP and caspase-12 were monitored by real-time RT-PCR and Western blot.To verify whether miR-21 was involved in the ERS-mediated apoptosis of the pulmonary fibroblasts,these cells were divided into three groups in random including the control group,miR-21 agomir group and miR-21 antagomir group.Both miR-21 agomir and miR-21 antagomir group were subjected to pretreat with NaHS at 40μmol/L at 60 min,whereafter receive TGF-β1 after transfected,followed by detection of the expression of GRP78,CHOP and caspase-12.Results:Compared with the control,the rate of cell proliferation and apoptosis significantly reduced in the H 2S protection group,similarly to the expression of miR-21.Compared with the TGF-β1 group,NaHS could obviously reduce the expression of GRP78,caspase-12 and CHOP.Compared with the negative group,the expression of GRP78,CHOP and caspase-12 significantly upregulated in miR-21 agomir group.However,the results was opposite in the miR-21 antagomir group.Conclusion:H 2S may inhibit pulmonary fibroblasts by regulating the expression of miR-21 and reducing ERS-mediated cell apoptosis.
作者 赵红 姚平波 ZHAO Hong;YAO Ping-Bo(School of Nursing of University of South China,Hengyang 421001,China)
出处 《中国免疫学杂志》 CAS CSCD 北大核心 2018年第11期1612-1616,共5页 Chinese Journal of Immunology
基金 本文为国家自然科学基金项目(No.81603108) 湖南省自然科学基金项目(2018JJ2339) 湖南省科卫联合项目(2018JJ6122) 湖南省教育科学规划资助项目(XJK17BGD070)。
关键词 硫化氢 肺成纤维化细胞 MICRORNA-21 内质网应激 凋亡 Hydrogen sulfide Pulmonary fibroblasts microRNA-21 ERS Apoptosis
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