IL-6对体外培养的牛眼小梁细胞中纤维连接蛋白的影响

Effect of IL-6 on fibronectin in bovine trabecular meshwork cells cultured in vitro

目的:探讨不同浓度白细胞介素-6(IL-6)刺激下体外培养的牛眼小梁细胞中纤维连接蛋白的表达变化。方法:采用组织块培养法取新鲜牛眼的小梁网组织,提取并培养第3代牛眼小梁细胞,采用细胞形态学对细胞进行鉴定。经终浓度为0、0.1、0.5、1ng/mL的IL-6药物刺激24h后,采用荧光定量PCR和蛋白质免疫印迹法检测各浓度IL-6刺激下牛眼小梁细胞中FNmRNA和蛋白的表达。结果:培养出的牛眼小梁细胞符合第3代牛眼小梁细胞形态特征。实时荧光定量PCR和蛋白质免疫印迹法显示,不同浓度IL-6刺激下的牛眼小梁细胞所产生的FNmRNA量分别为1.000±0.000、0.213±0.004、0.056±0.001、0.019±0.002,FN蛋白表达量分别为1.167±0.012、0.662±0.009、0.238±0.011、0.061±0.011,均呈下调趋势(rs=-0.713、-0.901,均P<0.05),4组间FNmRNA和蛋白表达均有差异(P<0.05)。结论:体外培养的牛眼小梁细胞在外源性IL-6刺激下影响FNmRNA和蛋白的表达,且IL-6浓度与蛋白表达呈负相关性,推测IL-6可能通过影响FN基因与蛋白的表达,进而改变小梁网组织结构。

AIM: To investigate the changes of fibronectin expression in cultured bovine trabecular meshwork cells when cells were stimulated by different concentrations of IL-6 to provide evidence for early diagnosis and new therapy of POAG. METHODS: We identify third-generation bovine trabecular meshwork cells which were got from tissue mass culture method. Then the relative expression of FN gene and protein in cells were detected by Real-Time PCR and Western-blot after 24h stimulation with 0ng/mL, 0.1ng/mL , 0.5ng/mL IL-6. RESULTS: The cultured bovine trabecular cells are coincident with what recorded in the book. Real-time PCR and Western blot showed that the amount of FN mRNA produced by cells was 1.000±0.000, 0.213±0.004, 0.056±0.001, 0.019±0.002 respectively, and the protein expression was 1.167±0.012, 0.662±0.009, 0.238±0.011, 0.061±0.011 respectively. There was a significant difference among four groups ( P <0.05). CONCLUSION: Cultured bovine trabecular meshwork cells have a negative correlation with the expression of FN protein after being stimulated by exogenous IL-6, and the results are consistent with the actual state of the disease. We speculate that the IL-6 participate in the pathogenesis and progression of POAG by affecting the expression of FN gene and protein and changing the structure of trabecular meshwork.