阿霉素肾病大鼠不同病理时期自噬体的形成及自噬相关蛋白的表达及分布情况及其与肾组织损伤、肾脏疾病进展的关系分析  

Autophagy formation and expression and distribution of autophagy-related proteins in adriamycin nephropathy rats at different pathological stages

作　　者：刘钰 曾光[1] 杨舸[2] LIU Yu;ZENG Guang;YANG Ge(Department of Rheumatology,Nejiang second people's Hospital,Neijiang,Sichuan,641000;Department of transformed Medicine,Neijiang second people's Hospital,Neijiang,Sichuan 641000)

机构地区：[1]内江市第二人民医院肾病风湿免疫科,四川内江641000 [2]内江市第二人民医院转化医学科,四川内江641000

出　　处：《临床和实验医学杂志》2019年第13期1350-1354,共5页Journal of Clinical and Experimental Medicine

基　　金：四川省医学会四川省医学科研项目(编号:S16043)

摘　　要：目的探讨阿霉素肾病(AN)大鼠自噬体在不同病理时期的形成和自噬相关蛋白表达、分布及其与大鼠肾脏疾病进展、肾组织损伤程度的相关性。方法将40只健康雄性SD大鼠随机分为对照组(于鼠尾静脉注入生理盐水)与实验组(于鼠尾静脉注入阿霉素6. 5 mg/kg),两组各20只。密切监测并比较两组大鼠15 d、30 d、45 d、60 d时血清尿素氮(BUN)、血肌酐(SCr)、总胆固醇(TC)、白蛋白(Alb)及24 h尿蛋白定量(24 h UPQ)水平的变化情况,观察两组大鼠足细胞结构和自噬体的形成及肾脏组织病理学变化情况。采用免疫荧光染色法对自噬相关蛋白Beclin-1、LC3-Ⅰ及LC3-Ⅱ在两组大鼠肾组织中的分布情况进行检测,并用Western蛋白印迹法对三种蛋白表达水平进行检测。结果实验组第15天时血清Alb明显降低,24 h UPQ显著增高,第30天时TC显著增高,与对照组相比存在显著性差异(P <0. 05);实验组第30天时血清BUN和SCr出现轻度增高,第45天时肾功能损伤加重,且出现进行性加重,与对照组相比存在显著性差异(P <0. 05)。光镜下观察实验组大鼠肾脏组织病理改变可见早期由微小病变肾病逐渐进展为局灶节段性肾小球硬化。透射电镜下观察大鼠早期存在系膜细胞增生,细胞器(如溶酶体等)分布和形态异常改变,足突增宽且出现弥漫性融合的现象,晚期观察可见足突消失和核固缩现象。免疫荧光染色结果可见对照组大鼠肾组织自噬能力较差,但在实验组中,随着肾脏疾病的进展,大鼠自噬能力进一步提高,且保持在较高水平。经Western蛋白印迹法检测结果所示,随着肾脏疾病的进展,实验组大鼠自噬相关蛋白Beclin-1、LC3-Ⅰ及LC3-Ⅱ蛋白表达明显提高,且与对照组相比存在显著性差异(P <0. 05)。结论随着AN病情程度的进展,大鼠肾功能损伤加重,且出现进行性加重,同时自噬能力进一步提高,自噬相关蛋白Beclin-1、LC3-Ⅰ及LC3-Ⅱ蛋�Objective To investigate the formation of autophages and the expression and distribution of autophagy-related protein in rats with adriamycin nephropathy (AN) at different pathological stages and their correlation with the progression of kidney disease and the degree of renal tissue injury in rats. Methods 40 healthy male SD rats were randomly divided into control group (injecting normal saline into rat tail vein) and experimental group (injecting adriamycin 6.5 mg/kg into rat tail vein),with 20 rats in each group. The changes of blood urea nitrogen (BUN),serum creatinine (SCr),total cholesterol (TC),albumin (Alb) and 24-hour urinary protein quantity (24-hour UPQ) levels were closely monitored and compared between the two groups at 15,30,45 and 60 days. The podocyte structure,autophagy formation and renal histopathological changes of the two groups were observed. The distribution of autophagy-related proteins Beclin-1,LC3-I and LC3-II in kidney tissues of rats in two groups was detected by immunofluorescence staining,and the expression levels of the three proteins were detected by Western blotting. Results Serum Alb decreased significantly at 15 days,24 h UPQ increased significantly,TC increased significantly at 30 days,and there was a significant difference compared with the control group ( P <0.05);serum BUN and SCR increased slightly at 30 days,renal function injury aggravated at 45 days,and progressively aggravated compared with the control group,there was a significant difference ( P <0.05). The histopathological changes of the kidneys in the experimental group were observed under light microscopy. It was found that the early stage of the disease progressed from minimal change nephropathy to focal segmental glomerulosclerosis. Under transmission electron microscopy,mesangial cell proliferation,abnormal distribution and morphology of organelles (such as lysosomes) were observed in the early stage of rats. The podocyte processes widened and diffuse fusion occurred. The podocyte processes disappeared and nuclea

关 键 词：大鼠 阿霉素肾病 自噬 自噬相关蛋白 肾组织损伤 疾病进展 

分 类 号：R692[医药卫生—泌尿科学]