膜性肾病尿蛋白对人肾小管上皮细胞凋亡的影响.docVIP

PAGE PAGE 5 膜性肾病尿蛋白对人肾小管上皮细胞凋亡的影响 冯胜军1，吴丽丹2,3，刘华锋2 （1广东医学院基础医学院微生物与免疫学教研室，广东 东莞 523808；2广东医学院附属医院肾病研究所，广东 湛江 524023；3广东省东莞市石龙人民医院肾内科，广东 东莞 523326） [摘 要] 目的 初步探讨膜性肾病尿蛋白对人肾小管上皮细胞凋亡存在剂量依赖关系。方法 提取膜性肾病患者尿蛋白，进行蛋白成份分析、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1.0mg/ml、2.0mg/ml、4.0 mg/ml、8.0 mg/ml蛋白浓度刺激人肾小管上皮细胞48h，空白组无蛋白，Hoechst33258染色法观察人肾小管上皮细胞凋亡情况。结果 膜性肾病尿蛋白由56.4％白蛋白、34.6％转铁蛋白、6.9％IgG组成。肌酐水平、内毒素均低于人体正常范围的下限。Hoechst33258染色法分析发现试验组与空白组之间，较大浓度尿蛋白组与较小浓度尿蛋白组之间人肾小管上皮细胞凋亡指数均有显著性差异（P0.05）。结论 肾小管上皮细胞凋亡程度与膜性肾病尿蛋白存在浓度依赖关系，本研究为深入揭示人类尿蛋白与肾小管-间质纤维化之间的机制提供重要依据。 [关键词] 膜性肾病；尿蛋白；肾小管上皮细胞；凋亡 Effects of urinary protein in patients with membranous nephropathy on apoptosis of human renal tubular epithelial cells FENG Sheng-jun1, WU Li-dan2,3, LIU Hua-feng2 1Department of Microbiology＆ Immunology，Guang Dong Medical College，Dongguan 523808，China;2Institute of Nephrology,Guangdong of Medical College, Zhanjiang 524001,China;3Nephrology department of Dongguan Shilong People’s Hospital, Dongguan 523326，China [Abstract]Objective：To study cell apoptosis on human renal tubular epithelial cells(hRTEC) induced by urinary protein from membranous nephropathy.Methods：Urinary protein in patients with membranous nephropathy was extracted, and analyzed of its for protein composition, bacterial endotoxin test and creatinine level. Test groups were different concentrations of urinary protein with 1.0mg/ml、2.0mg/ml、4.0 mg/ml、8.0 mg/ml respectively stimulating hRTEC for 48h,control group was no protein. hRTEC apoptosis index were detected by Hoechst33258 dying method to determine the influence of different concentration of urinary protein concentrations on hRTEC induced apoptosis. Results：Urinary protein of membranous nephropathy were consisted of 56.4% albumin, 34.6%transferrin, 6.9%IgG.and its endotoxin and creatinine were below the lower limit of normal range of human. Test groups and the control group there was significant difference(p0.05), and the larger group and the smaller the concentration of urinary protei