血管内皮生长因子C对大鼠近端肾小管上皮细胞凋亡的保护作用

Preventive effects of vascular endothelial growth factor-C on apoptosis of rat renal tubular epithelial cells

目的 探讨肿瘤坏死因子α（TNF-α）对肾小管上皮细胞生成血管内皮生长因子C（ VEGF-C）的影响,及VEGF-C对TNF-α诱导的肾小管上皮细胞凋亡的作用.方法 体外培养大鼠近端肾小管上皮细胞（NRK52E）,加入不同浓度的TNF-α作用不同时间,用实时定量PCR和Western blot方法检测VEGF-C mRNA和蛋白的表达.用不同浓度TNF-α与放线菌素D（ActD）刺激细胞24 h,流式细胞学方法检测细胞凋亡.再加入不同浓度的VEGF-C,观察其对凋亡的抑制情况.结果 TNF-α促进NRK52E细胞VEGF-C的表达,呈剂量和时间依赖性.TNF-α（加入ActD 30ng/ml）促进细胞凋亡,凋亡率随TNF-α浓度增高而增加.VEGF-C能够抑制凋亡,50 ng/ml时对细胞凋亡有抑制作用,100 ng/ml时抑制作用最强.结论 VEGF-C对大鼠近端肾小管上皮细胞的凋亡具有保护作用.

Objective To investigate the role of TNF-α in the production of VEGF-C and the impact of VEGF-C on the TNF-a/ActD-induced apoptosis of renal tubular epithelial cells. Methods NRK52E cells were cultured with different concentrations of TNF-α and different durations. Real-time PCR and Western blotting were used to examine the mRNA and protein expression of VEGF-C. After treatment with TNF-α/ActD for 24 h, cell apoptosis was detected by flow cytometry. Different concentrations of VEGF-C were added to evaluate its protective role in cell apoptosis. Results After treatment with TNF-α, the expression of VEGF-C was increased in a dose- and time-dependent man- ner. TNF-α combined with ActD could promote apoptosis in a dose-dependent manner. The apoptosis could by inhibited by VEGF-C. Conclusions VEGF-C could protect NRK52E ceils from apoptosis.