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目的模拟糖尿病条件,观察人肾小管上皮细胞(HK-2)生长情况和Smad1表达的变化,探讨高糖对肾小管上皮细胞的损伤机制。方法用DMEM/F12细胞液培养HK-2细胞,分为对照组,中糖组(12.5mmol/L葡萄糖),高糖组(25mmol/L葡萄糖),分别培养24h、48h、72h后倒置显微镜下观察HK-2的形态和数量变化,采用蛋白印迹技术检测Smad1的蛋白表达。结果高糖组刺激72h后,与对照组相比HK-2细胞数为(105.333±6.429)×105/m L,而对照组HK-2细胞数为(50.333±2.51)×105/m L,差异有统计学意义(P<0.01);高糖组HK-2细胞形态改变,Smadl蛋白表达明显增加。结论高糖能够诱导肾小管上皮细胞数量及形态发生改变,并且这一作用可能与Samd1表达增加有关。
Objective To simulate the condition of diabetes mellitus, observe the growth of human renal tubular epithelial cells (HK-2) and the expression of Smad1, and explore the mechanism of high glucose on renal tubular epithelial cells. Methods HK-2 cells were cultured in DMEM / F12 cells and divided into control group, moderate glucose group (12.5mmol / L glucose) and high glucose group (25mmol / L glucose), cultured for 24h, 48h and 72h respectively. The morphological and quantitative changes of HK-2 were observed and the protein expression of Smad1 was detected by Western blotting. Results Compared with the control group, the number of HK-2 cells was (105.333 ± 6.429) × 105 / m L and the number of HK-2 cells in the control group was (50.333 ± 2.51) × 105 / m L after 72 h of stimulation with high glucose. (P <0.01). The morphological changes of HK-2 cells in high glucose group and the increase of Smadl protein expression were observed. Conclusion High glucose can induce changes of renal tubular epithelial cell number and morphology, and this effect may be related to the increased expression of Samd1.