近视眼相关Lumican基因突变对人巩膜成纤维细胞中bFGF和TGF-β2的影响  

Regulation of bFGF and TGF-β2 in human scleral fibroblasts by the lumican gene mutation associated with myopia

作　　者：吕晓彤 宋彦铮[1] 张丰菊[1] Lyu Xiaotong;Song Yanzheng;Zhang Fengju(Beijing Tongren Eye Center,Beijing Tongren Hospital of Capital Medical University,Beijing Key Laboratory of Ophthalmology&Visual Sciences,Beijing 100730,China)

机构地区：[1]首都医科大学附属北京同仁医院北京同仁医院眼科中心,北京市眼科学与视觉科学重点实验室,100730

出　　处：《中华眼科杂志》2021年第4期277-283,共7页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金(81873682,81271041);北京市卫生系统高层次卫生技术人才学科带头人基金(2013-2-023);首都医科大学附属北京同仁医院青年人才培养计划种子基金(2018-YJJ-ZZL-029)。

摘　　要：目的探讨腺病毒介导的Lumican基因突变(c.596T>C)对人巩膜成纤维细胞(HSF)中碱性成纤维细胞生长因子(bFGF)和转化生长因子β2(TGF-β2)的影响,明确该突变在近视眼发生和发展中的作用。方法实验研究。应用组织块培养法分离培养HSF。取3~5代细胞转导病毒,转导突变型Lumican(c.596T>C)腺病毒的HSF为突变组、转导野生型Lumican腺病毒的HSF为野生组、转导缺陷型腺病毒的HSF为阴性对照组,未转导病毒的HSF为空白对照组。每组重复3次。采用实时荧光聚合酶链式反应(qPCR)检测各组细胞Lumican、bFGF、TGF-β2基因的表达水平。对比相同时间各组间的基因相对表达差异,差异以倍数变化法表示,采用单因素方差分析联合LSD-t检验进行统计学分析。结果突变组和野生组的Lumican基因表达量分别为空白对照组的103.146和398.646倍,高于阴性对照组,差异有统计学意义(t=-16.641,-21.729;P<0.05),阴性对照组与空白对照组差异无统计学意义(t=1.689,P>0.05)。突变组的bFGF基因表达量为空白对照组的2.812倍,TGF-β2基因表达量为空白对照组的2.346倍,均高于野生组及阴性对照组,差异有统计学意义(t=-3.921,-4.851;P<0.05),野生组、阴性对照组及空白对照组间差异无统计学意义(P>0.05)。结论Lumican基因突变(c.596T>C)使HSF中bFGF和TGF-β2的表达增加,提示Lumican基因突变(c.596T>C)可能通过调控bFGF和TGF-β2的表达改变巩膜组织的细胞外基质代谢,从而参与近视眼发生和发展过程中的巩膜重塑。Objective To investigate the regulation of basic fibroblast growth factor(bFGF)and transforming growth factor(TGF)-β2 in human scleral fibroblasts(HSFs)by the adenovirus-mediated Lumican gene mutation,and to illustrate the effect of this mutation on myopia.Methods Experimental study.The HSFs were isolated and cultured from human scleral tissues.The 3rd to 5th generation HSFs were transduced with Lumican mutant(c.596T>C)adenovirus,Lumican wild-type adenovirus,and defective adenovirus as the mutant group,wild group,and negative control group,respectively.Untransduced HSFs were defined as control group.The operation was conducted three times in each group.The expression levels of Lumican,bFGF and TGF-β2 were detected by qPCR.Statistical analysis of gene expression differences between groups was performed by fold changes.The differences were analyzed by one way ANOVA combined with LSD-t test.Results The expressions of Lumican in the mutant group and the wild group were 103.146-fold and 398.646-fold increased compared to the control group with significant difference(t=-16.641,-21.729;P<0.05).There was no statistical difference between the negative control group and the control group(t=1.689,P>0.05).The expressions of bFGF and TGF-β2 in the mutant group were 2.812-fold and 2.346-fold increased compared to the control group with significant difference,and higher than the other groups(t=-3.921,-4.851;P<0.05).There was no significant difference among the wild group,negative control group and control group(P>0.05).Conclusions The Lumican mutation(c.596T>C)increased the expressions of bFGF and TGF-β2 in HSFs.It indicates that the Lumican mutation(c.596T>C)may change the metabolism of extracellular matrix in the sclera by regulating bFGF and TGF-β2 to participate in scleral remodeling during the process of myopia.

关 键 词：近视 光蛋白聚糖 突变 巩膜 成纤维细胞生长因子2 转化生长因子Β2 

分 类 号：R778.11[医药卫生—眼科]