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目的:探讨胰岛素样生长因子-1(IGF-1)诱发离体培养的白内障患者晶状体上皮分泌趋化性细胞因子(MCP-1)的作用。方法:将白内障手术中分离糖尿病性白内障患者晶状体上皮和非糖尿病性白内障患者晶状体上皮细胞分别离体培养。加入重组IGF-1作用后,以RT-PCR方法检测MCP-1mRNA,夹心ELISA法检测MCP-1蛋白分泌水平。进行核提取后以Westernblot检测转录因子Sp-1核转移水平。利用Sp-1特异性阻抑剂米拉霉素共同孵育后收集上清观察MCP-1水平。结果:成功建立晶状体上皮细胞体外培养体系。IGF-1诱导伴发糖尿病性白内障患者晶状体上皮分泌MCP-1水平明显高于无糖尿病组。IGF-1作用于晶状体上皮细胞后能够活化Sp-1,Sp-1的特异性抑制能够明显降低MCP-1的表达。结论:IGF-1通过Sp-1途径诱导MCP-1的分泌表达。与非糖尿病性白内障患者相比,糖尿病性白内障患者晶状体上皮细胞产生更多的MCP-1。
Objective: To investigate the role of chemokine MCP-1 secreted by lens epithelial cells induced by insulin-like growth factor-1 (IGF-1) in cataract patients. Methods: The lens epithelial cells of patients with cataract and non-diabetic cataract isolated from cataract surgery were separately cultured. After addition of recombinant IGF-1, MCP-1 mRNA was detected by RT-PCR and the secretion of MCP-1 by sandwich ELISA. After nuclear extraction, the level of Sp-1 nuclear translocation was detected by Western blot. MCP-1 levels were observed by collecting the supernatants after incubation with sub-specific Sprague-Dawley (MDR) mithramycin. Results: The lens epithelial cells culture system was established successfully. IGF-1-induced diabetic cataract patients with lens epithelial secretion of MCP-1 was significantly higher than the non-diabetic group. The specific inhibition of Sp-1 can significantly reduce the expression of MCP-1 by IGF-1 activation of Sp-1 after lens epithelial cells. Conclusion: IGF-1 induces the secretion of MCP-1 through the Sp-1 pathway. Compared with non-diabetic cataract patients, lens epithelial cells in diabetic cataract produced more MCP-1.