足细胞自噬在膜性肾病经典动物模型中的作用及其激活可能的细胞内机制研究  

ROLE OF AUTOPHAGY IN THE CLASSIC ANIMAL MODEL OF MEMBRANOUS NEPHROPATHY AND ITS POSSIBLE MECHANISM OF CELL ACTIVATION

作　　者：鲍丽[1] 么冬爱[1] Bao Li;Yao Dongai(Physical Examination Center,Central South Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院体检中心

出　　处：《中国煤炭工业医学杂志》2019年第3期291-295,共5页Chinese Journal of Coal Industry Medicine

基　　金：湖北省自然科学基金资助项目(编号:2014CFB748)

摘　　要：目的探讨足细胞自噬在膜性肾病经典动物模型中的作用及其激活可能的细胞内机制。方法将50只SPF级健康雄性SD大鼠随机平均分为被动肾炎(PHN)2d组、PHN4d组、PHN7d组、PHN21d组以及对照组,建立大鼠PHN模型。观察足细胞形态的变化和自噬泡,并计算单个肾小球足细胞的数量。采用免疫组织化学染色法对肾小球内膜攻击复合物C5b-9的沉淀和半胱氨酸天冬氨酸蛋白酶(caspase)-3的表达进行检测,用TUNEL法对肾小球内细胞凋亡进行检测,Western印迹法对肾小球自噬相关蛋白LC3、GRP78、p-PERK、p-JNK、ATF6α的表达进行检测。结果至注射抗体后第21天可见典型模性肾病改变,而其尿蛋白量自建模3d起有显著上升,至21 d可达到(51.4±5.9)mg/24h。P62蛋白并无在PHN模型大鼠肾小球内发生堆积,故可确定LC3Ⅱ表达的提高非自噬流受阻导致。可观察到造模早期ATF6α、p-PERK、p-JNK有逐步上升的趋势,但21 d(即后期)又开始回落。结论膜性肾病免疫介导的损伤可通过激活内质网应激从而增强足细胞自噬作用以保持其内环境的稳定及生存。Objective To explore the role of autophagy in the classic animal model of membranous nephropathy and its possible mechanism of cell activation.Methods Fifty healthy maleSD rats of SPF grade were randomly and averagely divided into PHN 2 d group,PHN4 d group,PHN 7 d group,PHN21 d group and control group,and the PHN model was established.The morphological changes of podocytes and autophagic vesicles were observed,and the number of podocytes in a single glomerulus was calculated.The expression of C5 b-9 in the renal tissue and the expression of caspase-3 was detected by immunohistochemistry staining.Apoptosis of glomerular cells was detected by TUNEL method,and expression of GRP78,p-PERK,p-JNK,p-JNK,ATF6α,and LC3 were detected by Western blot.Results Typical model nephropathy was observed on the 21 st day after injection of antibody,and the urinary protein level increased significantly from the 3 d after injection,reaching(51.4+5.9)mg/24 h on the 21 st day.P62 protein did not accumulate in the glomeruli of PHN model rats,so it can be determined that the increase of LC3 II expression is caused by non-autophagic flow blockage.It can be observed that ATF6 a,p-PERK and p-JNK increased gradually in the early stage of modeling,but began to decline again in 21 days(i.e.later stage).Conclusion Immune-mediated injury in membranous nephropathy can enhance podocyte autophagy by activating endoplasmic reticulum stress to maintain the stability of its internal environment and survival.

关 键 词：内质网应激 足细胞 自噬 肾小球肾炎 膜性 

分 类 号：R692[医药卫生—泌尿科学]