不同病理类型肾病综合征患者尿蛋白对人肾小管上皮细胞凋亡的影响

Effects of Proteinuria on Apoptosis of Human Renal Tubular Epithelial Cells in Patients with Different Pathological Types of Renal Syndrome

目的探讨不同病理类型肾病综合征患者尿蛋白对人肾小管上皮细胞(hrTESc)凋亡的影响。方法提取人类微小病变肾病综合征、膜性肾病综合征、局灶-节段性肾病综合征患者尿蛋白,进行蛋白成分分析、细菌内毒素试验、蛋白溶液的肌酐水平等质量鉴定后,试验组分别按1、2、4、8 g/L尿蛋白浓度刺激hrTECs 48 h,空白组无尿蛋白,Hoechst33258染色法观察hrTECs凋亡情况。结果尿蛋白组分构成:微小病变肾病综合征清蛋白83.8%、转铁蛋白11.9%、IgG 3.4%;膜性肾病综合征清蛋白56.4%、转铁蛋白34.6%、IgG 6.9%;局灶-节段性肾病综合征清蛋白59.9%、转铁蛋白15.3%、IgG 13.8%。尿蛋白浓度1、2、4、8 g/L时,膜性肾病综合征组较微小病变肾病综合征组细胞凋亡率均升高(P<0.05);局灶-节段性肾病综合征组较膜性肾病综合征组和微小病变肾病综合征组细胞凋亡率均升高(P<0.05)。3组尿蛋白浓度1、2、4、8 g/L时较空白时,2、4、8 g/L时较1 g/L时,4、8 g/L时较2g/L时,8 g/L时较4 g/L时细胞凋亡率均升高(P<0.05)。结论 hrTECs的异常凋亡与尿蛋白具有浓度依赖关系,同时可能与尿蛋白中IgG成分密切相关,而尿清蛋白可能不是最重要的致病因子。

Objective To explore the effects of proteinuria on the apoptosis of human renal interstitial cells in patients with different pathological types of renal syndrome. Methods Urinary protein was extracted from patients with minimal change disease（MCD）,membranous nephropathy（MN）and focal-segmental renal synthesis（FSGS）. The protein composition analysis,bacterial endotoxin test and protein creatinine level test were conducted. The urinary protein with concentration of 1,2, 4 and 8 g/L respectively were used to stimulate human renal tubular epithelial cells（ hrTECs）for 48 h,and the control group was without urinary protein. The results were observed by Hoechst 33258 staining. Results Composition of urinary protein：MCD：albumin 83. 8%,transferrin 11. 9% and IgG 3. 4%;MN：albumin 56. 4%,transferrin 34. 6% and IgG 6. 9%;FSGS：albumin 59. 9%,transferrin 15. 3% and IgG 13. 8%. At different concentrations of urinary protein at 1,2,4 and 8 g/L,the apoptosis rate in MN group was higher than MCD group（P〈0. 05）. The apoptosis rate in FSGS group was higher than both the MN group and MCD group（P〈0. 05）. In addition,dose-dependent relationship existed in same pathological types of renal syndrome（P〈0. 05）. Conclusion There is dose-dependent relationship between the degree of apoptosis in hrTECs and the concentration of urine protein,which may be closely related to the IgG in urinary protein. However,the urinary albumin may not be the most crucial pathogenic factor.