独立生长因子1负调控GREM1基因抑制肝星状细胞增殖与活化实验研究  

Growth factor independence 1 negatively regulates GREM1 gene to inhibit proliferation and activation of hepatic stellate cells

作　　者：刘刚[1] 朱冬梅 赵芹[2] 刘晓玲 彭洪 LIU Gang;ZHU Dongmei;ZHAO Qin;LIU Xiaoling;PENG Hong(Department of Ultrasound,Nanchong Central Hospital,Nanchong 637000,China)

机构地区：[1]南充市中心医院,川北医学院第二临床医学院超声科,四川南充637000 [2]川北医学院,四川南充637000 [3]南充市中心医院普外科,四川南充637000

出　　处：《陕西医学杂志》2023年第6期655-659,共5页Shaanxi Medical Journal

基　　金：四川省中医药管理局科研专项课题(2020JC0078);四川省南充市市校合作科研专项基金资助项目(22SXQT0063)。

摘　　要：目的:探讨独立生长因子1(GFI1)负调控GREM1基因对肝星状细胞(HSC)增殖与活化的影响。方法:选择HSC细胞株LX-2培养并进行相关实验。采用CCK-8法检测GREM1基因和GFI1对LX-2细胞增殖的影响。采用Western blot分析GREM1基因和GFI1对α-平滑肌肌动蛋白(α-SMA)蛋白表达的影响。利用生物信息学方法分析GREM1基因的转录因子并通过染色质免疫共沉淀(CHIP)实验验证GREM1基因是否可与GFI1结合。采用实时荧光定量PCR(qRT-PCR)检测GFI1对GREM1 mRNA的影响。通过抑制GFI1并下调GREM1表达分析GFI1是否通过负调控GREM1抑制LX-2细胞增殖和α-SMA表达。结果:过表达GREM1可促进LX-2细胞增殖及上调α-SMA蛋白表达水平(均P<0.05)。GFI1可能是GREM1的上游转录因子,CHIP实验证实GFI1可与GREM1直接结合。下调GFI1表达可促进GREM1 mRNA表达和LX-2细胞增殖,并上调α-SMA蛋白表达水平(均P<0.05)。下调GREM1表达可以减弱GFI1表达(P<0.05)。结论:GREM1可促进HSC活化,而GFI1可通过负调控GREM1基因抑制HSC增殖与活化。Objective:To investigate the effect of growth factor independence 1(GFI1)on the proliferation and activation of hepatic stellate cells(HSC)by negative regulation of GREM1 gene.Methods:HSC cell line LX-2 was selected for culture and related experiments.The effects of GREM1 and GFI1 on the proliferation of LX-2 cells were detected by CCK-8 assay.The effects of GREM1 and GFI1 on the expression ofα-SMA protein were analyzed by Western blot.Bioinformatics method was used to analyze the transcription factors of GREM1 gene,and CHIP assay was used to verify whether GREM1 gene could bind to GFI1.The effect of GFI1 on GREM1 mRNA was detected by qRT-PCR.The inhibition of GFI1 and down-regulation of GREM1 were used to analyze whether GFI1 negatively regulated GREM1 to inhibit the proliferation andα-SMA expression of LX-2 cells.Results:Overexpression of GREM1 promoted the proliferation of LX-2 cells and up-regulated the expression ofα-SMA protein(all P<0.05).GFI1 might be an upstream transcription factor of GREM1,and CHIP assay confirmed that GFI1 could directly bind to GREM1.Down-regulation of GFI1 promoted GREM1 mRNA expression and proliferation of LX-2 cells,and up-regulatedα-SMA protein expression(all P<0.05).Down-regulation of GREM1 expression reduced the expression of GFI1(P<0.05).Conclusion:GREM1 can promote the activation of HSC,while GFI1 can inhibit the proliferation and activation of HSC by negatively regulating GREM1 gene.

关 键 词：肝星状细胞 独立生长因子1 GREM1基因 增殖 活化 机制 

分 类 号：R575.2[医药卫生—消化系统]