氯化锂通过TGFBI促进角膜基质成纤维细胞增殖和自噬

Lithium chloride promotes the proliferation and autophagy of corneal stromal fibroblasts by TGFBI

摘要:

目的：研究TGFBI和微管相关蛋白轻链3(LC3)在角膜营养不良患者中的表达，及氯化锂(LiCl)通过TGFBI对角膜基质成纤维细胞增殖能力的影响。

方法：用免疫组化和Western-blot方法检测角膜营养不良及正常角膜组织中TGFBI和LC3的表达。实验构建了TGFBI过表达载体并转染角膜基质成纤维细胞，分别以5、10、20、40mmol/L LiCl作用于突变型TGFBI转染的角膜基质成纤维细胞，检测不同时间(0、1、6、12h)后，TGFBI与LC3蛋白表达变化，并用CCK-8法检测细胞增殖活性。

结果：TGFBI和LC3在角膜营养不良患者角膜组织中显著高表达。TGFBI过表达抑制角膜基质成纤维细胞增殖活性(P<0.05)。LiCl抑制突变型TGFBI转染的角膜基质成纤维细胞中TGFBI和LC3蛋白表达，并增强其细胞增殖活性(P<0.05)。

结论：LiCl可以促进角膜基质成纤维细胞增殖活性和自噬，其作用机制与下调TGFBI和LC3的表达有关。

Abstract:

AIM: To study the expressions of TGFBI and microtubule-associated protein 1 light chain 3 alpha(LC3)in granular corneal dystrophy, and the influences of lithium chloride(LiCl)on corneal stromal fibroblast cell proliferation by TGFBI.

METHODS: Immunohistochemistry and Western-blot assays were used to detect the expression levels of TGFBI and LC3 in corneal dystrophy and normal corneal tissues. TGFBI overexpression vector was transfected into corneal stromal fibroblasts, and then the cells were treated with 5, 10, 20, 40mmol/L LiCl for different times(0, 1, 6, 12h), and Western-blot assay was performed to evaluate the expression levels of TGFBI and LC3, and CCK-8 assay was carried out to assess cell proliferation activity.

RESULTS: TGFBI and LC3 were highly expressed in corneal tissues of patients with corneal dystrophy. TGFBI overexpression inhibited the proliferation ability of corneal stromal fibroblasts(P<0.05). LiCl inhibited the expression levels of TGFBI and LC3, and enhanced the cell proliferation activity in corneal stromal fibroblasts transfected with mutant TGFBI(P<0.05).

CONCLUSION: LiCl promoted the proliferation and autophagy of corneal stromal fibroblasts, and its mechanism may be related to down regulated expressions of TGFBI and LC3.