Shh在糖尿病肾病患者肾组织中的表达及对肾小管上皮细胞凋亡的影响研究

Expression of Shh in renal tissue of patients with diabetic nephropathy and its effect on apoptosis of renal tubular epithelial cells

目的探讨音猥因子(Shh)在糖尿病肾病患者肾组织中的表达水平及对肾小管上皮细胞凋亡的影响。方法Western blot检测糖尿病肾病患者肾组织和肾癌患者正常肾组织中Shh的水平。实验分为5组,依次为低糖组、对照组、PCDNA3.1组、PCDNA3.1-Shh组、抑制剂组。低糖组用葡萄糖浓度为5.8 mmol/L的细胞培养液培养细胞,PCDNA3.1组、PCDNA3.1-Shh组、抑制剂组、对照组均用葡萄糖浓度为30 mmol/L的细胞培养液培养细胞,PCDNA3.1-Shh组细胞转染Shh过表达载体PCDNA3.1-Shh,PCDNA3.1-Shh组细胞转染空载体PCDNA3.1,抑制剂组细胞与JNK信号通路抑制剂作用。Western blot检测对照组、PCDNA3.1组、PCDNA3.1-Shh组细胞中Shh水平。流式细胞术检测各组细胞凋亡情况。Western blot检测各组细胞中Bcl-2、Bax、JNK、p-JNK表达水平。结果 Shh在糖尿病肾病患者肾组织中的表达水平明显低于正常肾组织(P<0.05)。PCDNA3.1-Shh组细胞中Shh表达水平明显高于对照组和PCDNA3.1组(P<0.05)。对照组细胞凋亡率明显高于低糖组(P<0.05)。PCDNA3.1-Shh组细胞凋亡率明显低于对照组和PCDNA3.1组(P<0.05)。对照组Bax和p-JNK水平明显高于低糖组,而Bcl-2水平明显低于低糖组(P<0.05)。PCDNA3.1-Shh组细胞Bax和p-JNK水平明显低于对照组,而Bcl-2水平明显高于对照组(P<0.05)。抑制剂组细胞凋亡趋势与PCDNA3.1-Shh组细胞相一致。结论 Shh在糖尿病肾病患者肾组织中表达下调。Shh能够明显改善高糖诱导的肾小管上皮细胞凋亡,作用机制与JNK信号通路有关。

Objective To explore the expression of Shh in renal tissue of patients with diabetic nephropathy and its effect on apoptosis of renal tubular epithelial cells.Methods The expression of Shh in renal tissue of patients with diabetic nephropathy and normal renal tissue in patients with renal carcinoma were detected by Western blot.The experiment was divided into 5 groups,followed by low glucose group and the control group,PCDNA3.1 group,PCDNA3.1-Shh group,inhibitor group,low glucose group cultured cells with glucose concentration of 5.8 mmol/L,PCDNA3.1 group,PCDNA3.1-Shh group,inhibitor group and control group cultured cells with glucose concentration cultured cells were30 mmol/L,PCDNA3.1-Shh group transfected with PCDNA3.1-Shh,PCDNA3.1-Shh group transfected with PCDNA3.1.Inhibitor group effected with JNK signaling pathway inhibitor.Western blot was used to detect the Shh levels in control group,PCDNA3.1 group,PCDNA3.1-Shh group.Cell apoptosis was detected by flow cytometry.Detected the levels of Bax,Bcl-2,JNK and p-JNK by Western blot.Results The expression level of Shh in renal tissues of patients with diabetic nephropathy was significantly lower than that of normal renal tissues(P < 0.05).The expression level of Shh in PCDNA3.1-Shh group was significantly higher than that in control group and PCDNA3.1 group(P < 0.05).The apoptosis rate of control group was significantly higher than that of the low sugar group(P < 0.05).The apoptosis rate of PCDNA3.1-Shh group was significantly lower than that of control group and PCDNA3.1 group(P < 0.05).The levels of Bax and p-JNK in the control group were significantly higher than those in the low sugar group,while the Bcl-2 level was significantly lower than that of the low sugar group(P < 0.05).The levels of Bax and p-JNK in PCDNA3.1-Shh group were significantly lower than those in the control group,while the Bcl-2 level was significantly higher than that in the low sugar group(P < 0.05).The apoptosis trend of the inhibitor group was consistent with that of the PCDNA3.1-Shh group.Conclusion Shh expression is down regulated in renal tissue of diabetic nephropathy patients.Shh can obviously improve the apoptosis of renal tubular epithelial cells induced by high glucose,and the mechanism of action is related to the JNK signaling pathway.