Smad泛素化调节因子2对人胚胎眼巩膜成纤维细胞TGF-β1/Smads信号通路和I型胶原表达的影响  

Effect of Smad ubiquitination regulator factor 2(Smurf 2)on TGF-β1/Smads signal pathway andexpression of type I collagen in human fetal scleral fibroblasts(HFSFs)cells

作　　者：陈婷[1] 陈丽娟[2] 陈银河 CHEN Ting;CHEN Lijuan;CHEN Yinhe(Department of Ophthalmology,the Second Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian 362000,China)

机构地区：[1]福建医科大学附属第二医院眼科,泉州362000 [2]漳州卫生职业学院医学技术系眼视光教研室 [3]泉州市妇幼保健院·儿童医院眼科

出　　处：《福建医药杂志》2022年第2期110-113,F0004,共5页Fujian Medical Journal

基　　金：福建省教育厅中青年教师教育科研项目(JAT170249)。

摘　　要：目的探讨Smad泛素化调节因子2(Smurf2)在转化生长因子-β1(TGF-β1)诱导后对人胚胎眼巩膜成纤维细胞(HFSFs)分泌Smads蛋白、I型胶原α1(COLIA1)的影响,探讨泛素化在巩膜重塑中的作用。方法HFSFs复苏并稳定传代后,免疫组化(IHC)法检测细胞中Smurf2的表达。将HFSFs细胞分别设空白对照组、TGF-β1组(加入10 ng/mL TGF-β1)、Control siRNA转染组(10 ng/mL TGF-β1+Control siRNA)及Smurf2 siRNA转染组(10ng/mL TGF-β1+Smurf2 siRNA)。Western Blot法检测COLIA1蛋白表达水平。RT-PCR法检测各组细胞中Smurf2、Smad2、Smad3和Smad7的mRNA表达水平。结果IHC结果显示,HFSFs中有Smurf2蛋白表达。TGF-β1诱导HFSFs中COLIA1蛋白显著性高表达,而当Smurf2被抑制时,COLIA1蛋白表达受抑制。与空白对照组比较,Smad7 mRNA表达水平呈上升趋势,差异有统计意义(P<0.05),但Smad7蛋白表达水平与PCR结果不完全相同;各培养组中Smad2和Smad3 mRNA和蛋白表达水平,与对照组比较,差异均无统计学意义(P>0.05)。结论Smurf2表达于HFSFs细胞中,且随着Smurf2表达减少,HFSFs细胞中Smad7蛋白含量呈上升趋势,COLIA1含量呈下降趋势。提示Smurf2可能通过作用于TGF-β1/Smads信号通路参与了巩膜重塑过程。Objective To explore the effect of Smad ubiquitination regulator factor 2(Smurf2)on the secretion of Smads protein and Collagen type Iα1(COLIA1)by human fetal scleral fibroblasts(HFSFs)after induction by transforming growth factor-β1(TGF-β1).Methods After recovery and stable passage of HFSFs,expression of Smurt2 in cells was detected by immunohistochemistry(IHC).The HFSFs were set up as control group,TGF-β1 group(10ng/mL TGF-β1),control siRNA transfection group(10 ng/mL TGF-β1+control siRNA)and Smurf2 siRNA transfection group(10 ng/mL TGF-β1+Smurf2 siRNA).Western Blot method was used to detect the expression level of COLIA1.The mRNA expression levels of Smurf2,Smad2,Smad3 and Smad7 in each group were detected by RT-PCR.Results IHC showed the expression of Smurf2 in HFSFs.The COLIA1 of HFSFs induced by TGF-β1 was highly expressed.When Smurf2 was inhibited,the expression of COLIA1 decreased significantly.Compared with the control group,the mRNA expression level of Smad7 showed an upward trend,and the difference was statistically significant(P<0.05).However,the Smad7 protein expression level was not exactly the same as the PCR result.Compared with the control group,the mRNA and protein expression levels of Smad2 and Smad3 in the other three groups were not statistically significant(P>0.05).Conclusion Smurf2 is expressed in HFSFs,and as the expression of Smurf2 decreases,the protein content of Smad7 in HFSFs shows an upward trend,while the content of COLIA1 shows a downward trend.The Smurf2 may be involved in the pathological process of scleral remodeling by acting on the TGF-β1/Smads signaling pathway.

关 键 词：Smad泛素化调节因子2(Smurf2) 人胚胎眼巩膜成纤维细胞(HFSFs细胞) TGF-β1/Smad通路 近视 

分 类 号：R778.1[医药卫生—眼科]