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目的:建立高糖诱导的肾小管上皮细胞转分化模型,检测肾小管上皮细胞转分化过程中Smad锚着蛋白(Smad anchor for receptor activation,SARA)的表达变化。方法:采用Western印迹和实时定量PCR检测细胞波形蛋白(vimentin)、紧密连接蛋白(Zona occludens-1,ZO-1)、SARA蛋白及其mRNA表达水平。结果:30mmol/LD-葡萄糖刺激后,HK-2细胞vimentin蛋白及其mRNA水平呈时间依赖性升高,而ZO-1蛋白及其mRNA水平呈时间依赖性下降,此变化在高糖刺激48h时最为显著。同时,SARA蛋白及其mRNA表达水平呈时间依赖性下降。结论:高糖可诱导HK-2细胞发生转分化,SARA可能作为保护因子参与了这一过程。
OBJECTIVE: To establish a model of renal tubular epithelial cell transdifferentiation induced by high glucose and to detect the expression of Smad anchor for receptor activation (SARA) during the process of renal tubular epithelial cell transdifferentiation. Methods: The expressions of vimentin, ZO-1 and SARA protein and their mRNA were detected by Western blotting and real-time PCR. Results: After stimulation with 30 mmol / L glucose, the vimentin protein and mRNA levels in HK-2 cells increased in a time-dependent manner, while the expression of ZO-1 protein and mRNA decreased in a time-dependent manner. The most significant. Meanwhile, the expression of SARA protein and its mRNA decreased in a time-dependent manner. Conclusion: High glucose can induce HK-2 cells to undergo transdifferentiation. SARA may participate in this process as a protective factor.