曲尼司特对青光眼患者体外培养的眼部Tenon囊成纤维细胞增殖和移行的影响

目的 探讨曲尼司特(tranilast)对青光眼患者体外培养的眼部Tenon囊成纤维细胞增殖及移行的影响。方法 取青光眼患者滤过术中剪下的Tenon囊组织,在体外进行成纤维细胞原代及传代培养。分别采用MTT法、细胞计数法、免疫组化加图像分析法及划线法,研究不同浓度的曲尼司特对体外培养的成纤维细胞增殖及移行的影响及其与蛋白激酶C(PKC)表达的关系。结果 当浓度在12．5～100．0mg／L之间变化时,曲尼司特能明显抑制成纤维细胞的增殖,强度呈剂量依赖性;50．0mg／L和100．0mg／L的曲尼司特能明显抑制成纤维细胞的移行,并下调细胞内PKC的表达。结论 曲尼司特能抑制成纤维细胞的增殖及移行,这种作用可能与下调细胞内PKC的表达有关。

The investigation of tranilast on the proliferation and migration of human Tenon's capsule fibroblasts

OBJECTIVE: To investigate the effect of tranilast, N-(3,4-dimethoxycinnamoyl) anthramilic acid, on the proliferation and migration of human Tenon's capsule fibroblasts. METHODS: fibroblasts were cultured from Tenon's tissue of a glaucoma patient after trabeculectomy. The subcultured cells were incubated with DMEM medium containing different concentrations of tranilast for 72 h. The growth of fibroblasts was measured by methyl thiazlyl tetrazolium (MTT) assay and the cell count, and migration was evaluated by crutch method. The expression level of Protein kinase C (PKC) in fibroblasts was tested by immunohistochemical associated with image biological analysis (IBAS) methods. RESULTS: Treated with tranilast varying from 12.5 mg/L to 100.0 mg/L concentration, the proliferation of fibroblasts declined in a dose dependent manner. The migration of fibroblasts decreased from 40.20 +/- 5.83 to 22.50 +/- 4.21 and 9.80 +/- 2.14 cells/per field (P < 0.05) at 50.0 and 100.0 mg/L, respectively, and PKC expression was suppressed by tranilast from 0.2591 +/- 0.0038 to 0.2375 +/- 0.0106 and 0.1273 +/- 0.0573 Absorption value (P < 0.05) at 50.0 and 100.0 mg/L, respectively. CONCLUSION: Tranilast inhibited the proliferation as well as migration of fibroblasts in vitro, at least in part, by downregulation of PKC expression.