Lyu X T, Song Y Z, Zhang F J
Beijing Tongren Eye Center, Beijing Tongren Hospital of Capital Medical University, Beijing Key Laboratory of Ophthalmology & Visual Sciences, Beijing 100730, China.
Zhonghua Yan Ke Za Zhi. 2021 Apr 11;57(4):277-283. doi: 10.3760/cma.j.cn112142-20200825-00553.
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. 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- test. 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 (=-16.641, -21.729; <0.05). There was no statistical difference between the negative control group and the control group (=1.689, >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 (=-3.921, -4.851; <0.05). There was no significant difference among the wild group, negative control group and control group (>0.05). 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. (.
通过腺病毒介导的亮氨酸富含蛋白聚糖(Lumican)基因突变,研究其对人巩膜成纤维细胞(HSFs)中碱性成纤维细胞生长因子(bFGF)和转化生长因子(TGF)-β2的调控作用,并阐明该突变对近视的影响。实验研究。从人巩膜组织中分离培养HSFs。将第3至5代HSFs分别用Lumican突变体(c.596T>C)腺病毒、Lumican野生型腺病毒和缺陷型腺病毒转导,作为突变组、野生组和阴性对照组。未转导的HSFs定义为对照组。每组进行3次操作。采用qPCR检测Lumican、bFGF和TGF-β2的表达水平。通过倍数变化对组间基因表达差异进行统计分析。采用单因素方差分析结合LSD检验分析差异。与对照组相比,突变组和野生组中Lumican的表达分别增加了103.146倍和398.646倍,差异有统计学意义(=-16.641,-21.729;<0.05)。阴性对照组与对照组之间无统计学差异(=1.689,>0.05)。与对照组相比,突变组中bFGF和TGF-β2的表达分别增加了2.812倍和2.346倍,差异有统计学意义,且高于其他组(=-3.921,-4.851;<0.05)。野生组、阴性对照组和对照组之间无显著差异(>0.05)。Lumican突变(c.596T>C)增加了HSFs中bFGF和TGF-β2的表达。这表明Lumican突变(c.596T>C)可能通过调节bFGF和TGF-β2改变巩膜细胞外基质的代谢,从而参与近视过程中的巩膜重塑。(.