CD36在糖尿病肾病患者肾组织中的表达及调控Wnt/β-catenin信号通路对人肾小管上皮细胞增殖凋亡的实验研究-东南大学学报医学版

CD36在糖尿病肾病患者肾组织中的表达及调控Wnt/β-catenin信号通路对人肾小管上皮细胞增殖凋亡的实验研究

单位：枣庄矿业集团中心医院内分泌科, 山东枣庄 277000

关键词：CD36基因糖尿病肾病人肾小管上皮细胞 Wnt/β-catenin信号通路增殖凋亡

分类号：Q255;R692.6

出版年·卷·期（页码）：2017·36·第五期（704-710）

摘要：

目的：探讨CD36在糖尿病肾病患者肾组织中的表达及调控Wnt/β-catenin信号通路对人肾小管上皮细胞增殖凋亡的影响。方法：Western blot检测CD36在糖尿病肾病中的表达；CD36小干扰RNA （CD36-siRNA）和阴性对照（siRNA-NC）转染人肾小管上皮细胞HKC，以空脂质体转染的细胞作为对照组，Western blot检测转染48 h后各组细胞中CD36蛋白表达；将后续实验分为低糖组、高糖组、siRNA-NC+高糖组、CD36-siRNA+高糖组，各实验组细胞培养48 h后，CCK8试验检测细胞增殖，流式细胞术检测细胞凋亡，Western blot检测B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl2-Associated X的蛋白质（Bax）、p53、β-连环蛋白（β-catenin）、细胞周期素D1（CyclinD1）蛋白表达。结果：CD36在糖尿病肾病的表达显著高于正常肾组织（P<0.01）；CD36-siRNA组CD36蛋白表达水平显著低于对照组（P<0.01）；高糖组细胞存活率及Bcl-2、β-catenin、CyclinD1蛋白表达显著低于低糖组，细胞凋亡率及Bax、p53蛋白表达显著高于低糖组（P<0.01），siRNA-NC+高糖组细胞存活率、细胞凋亡率及Bcl-2、Bax、p53、β-catenin、CyclinD1蛋白表达与高糖组比较差异无统计学意义（P>0.05）；CD36-siRNA组细胞存活率及Bcl-2、β-catenin、CyclinD1蛋白表达显著高于高糖组，细胞凋亡率及Bax、p53蛋白表达显著低于高糖组（P<0.01）。结论：CD36在糖尿病肾病中高表达，高糖能抑制人肾小管上皮细胞HKC的增殖，促进细胞凋亡，而沉默CD36基因的表达能减弱这种效果，其机制与Wnt/β-catenin信号通路的调控有关。

Objective:To investigate the expression of CD36 in diabetic nephropathy and regulation of Wnt/β-catenin signaling experimental study on proliferation and apoptosis of human renal tubular epithelial cells. Methods:The expression of CD36 in diabetic nephropathy was detected by Western blot; CD36 small interfering RNA (CD36-siRNA) and negative control (siRNA-NC) were transfected into human renal tubular epithelial cells HKC, and empty liposome transfected cells as the control group; the expression of CD36 protein after transfected for 48 h was detected by Western blot; the subsequent experiments were divided into low glucose group, high glucose group, siRNA-NC+high glucose group, and CD36-siRNA+high glucose group, the experimental group cell were cultured for 48 h, cell proliferation was detected by CCK8 test, apoptosis was detected by flow cytometry, the expression of p53, Bax, Bcl-2, β-catenin and CyclinD1 protein were detected by Western blot. Results:The expression of CD36 in diabetic nephropathy was significantly higher than that in normal renal tissue (P<0.01); the expression level of CD36 protein in CD36-siRNA group was significantly lower than the control group (P<0.01); cell survival rate and Bcl-2, β-catenin and CyclinD1 protein expression in high glucose group was significantly lower than in low glucose group, while the apoptosis rate and the expression of Bax and p53 protein was significantly higher than low sugar group (P<0.01); cell survival rate, apoptosis rate and Bcl-2, Bax, p53, β-catenin and CyclinD1 protein expression in siRNA-NC+high glucose group compared with high glucose group had no significant difference (P>0.05); the survival rate and Bcl-2, β-catenin and CyclinD1 protein expressionin in CD36-siRNA+high glucose group was significantly higher than in high glucose group, while the expression of Bax and p53 protein and the apoptosis rate was significantly lower than the high glucose group (P<0.01). Conclusion:The expression of CD36 in diabetic nephropathy is higher, high glucose can inhibit cell proliferation of human renal tubular epithelial cells HKC, promote cell apoptosis, and Silencing the expression of the CD36 gene could attenuate this effect, and its mechanism is related to the regulationof Wnt/β-catenin signaling pathway.

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