CAJ | 학술논문

目的探讨内质网应激在慢性蛋白尿模型大鼠肾小管上皮细胞凋亡中的作用及赖诺普利的干预效果.方法 18只雄性Wistar大鼠随机分为正常对照组(正常组,n=6)、慢性蛋白尿模型组(模型组,n=6)和赖诺普利治疗组(治疗组,n=6),模型组和治疗组一次性尾静脉注射阿霉素2.0 mg/kg诱导慢性蛋白尿模型,正常组注射等量生理盐水.造模后第1天开始,治疗组予赖诺普利10 mg· kg-1·d-1灌胃,正常组和模型组予等量蒸馏水灌胃,共12周.留取各组大鼠第4、8、12周24 h尿标本行尿总蛋白检测,留取12周时各组大鼠血清及肾脏组织标本,检测肾功能、血清总蛋白及白蛋白等生化指标,使用TUNEL方法检测肾小管上皮细胞凋亡,免疫组化及Western印迹方法检测肾组织葡萄糖调节蛋白78 (GRP78)、CCAAT增强子结合蛋白同源蛋白(CHOP)表达.结果第4、8、12周时,与正常组相比,模型组大鼠24 h尿蛋白量明显升高(P＜0.05),治疗组较模型组有明显降低(P＜0.05).第12周时,与正常组相比,模型组血清白蛋白明显降低(P＜0.05),血清总蛋白、血肌酐、血尿素氮变化差异无统计学意义,TUNEL阳性肾小管上皮细胞明显增多(P＜0.05),GRP78、CHOP表达明显上调(P＜0.05).与模型组相比,治疗组血清白蛋白明显升高(P＜0.05),血清总蛋白、血肌酐、血尿素氮变化差异无统计学意义,TUNEL阳性肾小管上皮细胞明显减少(P＜0.05),GRP78、CHOP表达明显下调(P＜0.05).结论慢性蛋白尿模型大鼠肾小管上皮细胞存在内质网应激激活,并可能通过促凋亡因子CHOP通路引起肾小管上皮细胞凋亡,赖诺普利可能通过抑制内质网应激信号通路减轻肾小管上皮细胞凋亡.
목적 탐토내질망응격재만성단백뇨모형대서신소관상피세포조망중적작용급뢰낙보리적간예효과.방법 18지웅성Wistar대서수궤분위정상대조조(정상조,n=6)、만성단백뇨모형조(모형조,n=6)화뢰낙보리치료조(치료조,n=6),모형조화치료조일차성미정맥주사아매소2.0 mg/kg유도만성단백뇨모형,정상조주사등량생리염수.조모후제1천개시,치료조여뢰낙보리10 mg· kg-1·d-1관위,정상조화모형조여등량증류수관위,공12주.류취각조대서제4、8、12주24 h뇨표본행뇨총단백검측,류취12주시각조대서혈청급신장조직표본,검측신공능、혈청총단백급백단백등생화지표,사용TUNEL방법검측신소관상피세포조망,면역조화급Western인적방법검측신조직포도당조절단백78 (GRP78)、CCAAT증강자결합단백동원단백(CHOP)표체.결과 제4、8、12주시,여정상조상비,모형조대서24 h뇨단백량명현승고(P＜0.05),치료조교모형조유명현강저(P＜0.05).제12주시,여정상조상비,모형조혈청백단백명현강저(P＜0.05),혈청총단백、혈기항、혈뇨소담변화차이무통계학의의,TUNEL양성신소관상피세포명현증다(P＜0.05),GRP78、CHOP표체명현상조(P＜0.05).여모형조상비,치료조혈청백단백명현승고(P＜0.05),혈청총단백、혈기항、혈뇨소담변화차이무통계학의의,TUNEL양성신소관상피세포명현감소(P＜0.05),GRP78、CHOP표체명현하조(P＜0.05).결론 만성단백뇨모형대서신소관상피세포존재내질망응격격활,병가능통과촉조망인자CHOP통로인기신소관상피세포조망,뢰낙보리가능통과억제내질망응격신호통로감경신소관상피세포조망.
Objective To investigate the role of endoplasmic reticulum stress in tubular epithelial cell apoptosis in chronic proteinuria rat model and the effect of lisinopril intervention.Methods Adriamycin nephropathy was induced in male Wistar rats (n=12) by a single injection of adriamycin at 2 mg/kg body weight.Rats were then randomly assigned to model group or treatment group,to which distilled water or lisinopril were administered respectively for 12 weeks.Six normal rats serving as controls were administered distilled water.24 h urine samples were collected at week 4,8,12 and the urine protein was measured.At the end of study,serum was obtained and physiological parameters (serum creatinine,urea,total protein and albumin) were measured.Renal tubular epithelial cell apoptosis was assessed by TUNEL.GRP78,CHOP protein expression in kidney was quantified by immunohistochemistry and Western blotting.Results Compared to control group rats,increased proteinuria was observed in model group rats at week 4,8,12 (P＜0.05).Lisinopril treatment attenuated urine protein excretion significantly (P ＜ 0.05).At week 12,hypoalbuminemia was detected in model group rats (P ＜ 0.05),whereas the condition was alleviated by lisinopril (P ＜ 0.05).There were no significant differences of serum creatinine,urea and total protein in each group (P ＞ 0.05).Compared to control group rats,increased TUNEL positive tubular epithelial cells and tubular GRP78 and CHOP expression were also observed in model group rats (P ＜ 0.05);however,these conditions in the kidney were significantly decreased in treatment group (P ＜ 0.05).Conclusions Endoplasmic reticulum stress may be involved in the process of tubular epithelial cell apoptosis induced by proteinuria.Lisinopril may attenuate tubular epithelial cell apoptosis through regulating this signal pathway.