ROCK抑制剂对青光眼术后瘢痕形成的作用机制

Mechanism of ROCK inhibitor on scar formation after glaucoma surgery

目的:探讨Rho相关卷曲螺旋形成蛋白激酶(ROCK)抑制剂对青光眼手术后瘢痕形成的影响,分析ROCK抑制剂对瘢痕修复的作用机制。方法:健康大鼠分为模型组和ROCK抑制剂组(RR组),采用MTT法测定瘢痕组织中成纤维细胞增殖情况,RT-PCR检测滤过道瘢痕组织转化生长因子-β1(TGF-β1)、α平滑肌肌动蛋白(α-SMA)、ROCK mRNA表达,免疫组织化学检测白细胞介素(IL)-1β(IL-1β)、α-SMA蛋白阳性表达,免疫印迹检测IL-1β、IL-6、血管内皮生长因子(VEGF)、α-SMA蛋白表达。结果:模型组成纤维细胞的抑制率低于RR组的抑制率,差异有统计学意义。RT-PCR检测结果显示模型组TGF-β1、ROCK、α-SMA mRNA的表达高于RR组。免疫组织化学检测结果显示RR组IL-1β、α-SMA的阳性表达显著低于模型组。免疫印迹检测结果显示模型组IL-1β、IL-6、VEGF、α-SMA的表达水平高于RR组,差异均有统计学意义。结论:ROCK抑制剂可通过降低TGF-β1 mRNA、ROCK mRNA、IL-1β、IL-6、VEGF、α-SMA蛋白的表达进而抑制成纤维细胞的增殖,阻碍青光眼术后瘢痕的形成。

Objective:To explore the effects of ROCK inhibitors on scar formation after glaucoma surgery,and analyze the mechanism of ROCK inhibitors on scar repair.Methods:Healthy rats were divided into the model group and the ROCK inhibitor group(RR group).The proliferation of fibroblasts in the scar tissue was measured by the MTT method.The expression of transforming growth factor-β1(TGF-β1),αsmooth muscle actin(α-SMA),and ROCK mRNA in the filter tract scar tissue was detected by RT-PCR.Immunohistochemistry was used to detect positive expression of interleukin(IL)-1β(IL-1β)andα-SMA protein,and immunoblotting was used to detect expression of IL-1β,IL-6,vascular endothelial growth factor(VEGF),andα-SMA protein.Results:The inhibition rate of fibroblasts in the model group was lower than that in the RR group,and the difference was statistically significant.RT-PCR assay showed that the expression of TGF-β1,ROCK,andα-SMA mRNA in the model group was higher than that in the RR group.Immunohistochemical assay results showed that the positive expression of IL-1βandα-SMA in the RR group was significantly lower than that in the model group.The results of immunoblotting showed that the expression levels of IL-1β,IL-6,VEGF,andα-SMA in the model group were higher than those in the RR group,and the above differences were statistically significant.Conclusion:ROCK can inhibit the proliferation of fibroblasts by reducing the expressions of TGF-β1 mRNA,ROCK mRNA,IL-1β,IL-6,VEGF,andα-SMA,thereby hindering the formation of postoperative scars of glaucoma.