CPT1A对糖尿病肾病肾间质纤维化的保护作用  

CPT1A protects against renal fibrosis in mouse model of diabetic kidney disease

作　　者：熊莉 赵景宏 何婷 肖堂利 刘赤 XIONG Li;ZHAO Jinghong;HE Ting;XIAO Tangli;LIU Chi(Department of Nephrology,PLA Center for Nephrology,Chongqing Key Laboratory for Prevention and Treatment of Chronic Kidney Diseases,Chongqing Clinical Research Center of Kidney and Urology Diseases,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China)

机构地区：[1]陆军军医大学(第三军医大学)第二附属医院肾内科,全军肾脏病中心,重庆市慢性肾脏病防治重点实验室,重庆市肾病与泌尿系统疾病临床医学研究中心,重庆400037

出　　处：《陆军军医大学学报》2023年第8期810-816,共7页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(82270762)。

摘　　要：目的探讨肉碱棕榈酰转移酶1A(palmitoyl-transferase 1A,CPT1A)在糖尿病肾病(diabetic kidney disease,DKD)肾间质纤维化中的作用。方法16只4周龄雄性C57BL/6小鼠随机分为(n=8):对照组(Ctrl组,给予普通饲料喂养)和DKD组[通过高脂饮食联合链脲佐菌素(streptozotocin,STZ)制备DKD模型]。每4周收集小鼠24 h尿液,检测空腹血糖和体质量。建模16周(小鼠20周龄)后取材并检测2组小鼠血肌酐水平、尿蛋白/尿肌酐比值,PAS和Masson染色观察肾小球病理改变和肾间质纤维化程度,免疫组化染色、Western blot观察CPT1A及纤维化标志物的表达。体外培养人近端肾小管上皮细胞(proximal tubular cell line,HK-2)后分3组:正常对照(Ctrl)组、30 mmol/L高糖(high glucose,HG)组、30 mmol/L甘露醇(mannitol,M)组,孵育48 h后观察CPT1A、纤维化标志物的表达。CPT1A过表达质粒转染HK-2细胞后再予以30 mmol/L高糖处理,观察各组细胞形态和纤维化标志物的表达。结果与Ctrl组小鼠相比,DKD组小鼠肾小球体积增大、系膜基质增多和出现肾间质纤维化,同时血肌酐、尿蛋白/尿肌酐比值显著升高(P<0.05),肾组织CPT1A表达明显降低,而纤维化标志物表达显著升高。体外实验中:高糖导致HK-2细胞失去原有形态、呈长梭形改变,CPT1A表达降低和纤维化标志物表达升高,并与渗透压改变无关。而CPT1A过表达能抑制高糖诱导的HK-2细胞形态改变和纤维化标志物高表达。结论上调CPT1A表达可能对糖尿病肾病肾间质纤维化具有潜在的抑制作用。ObjectiveTo determine the effect of carnitine palmitoyl-transferase 1A(CPT1A)on renal interstitial fibrosis in diabetic kidney disease(DKD).MethodsA total of 164-week-old male C57BL/6 mice were randomly divided into Control(Ctrl)group(n=8)and DKD group(n=8).The mice of Ctrl group were fed with normal diet,and those from the DKD group were inflicted with intraperitoneal streptozotocin injection plus high fat diet(STZ/HFD)to induce DKD model.Their 24-hour urine were collected,and fasting blood glucose level and body weight were measured every 4 weeks.After 16 weeks of modeling,the mice(aged 20 weeks)were sacrificed,and the levels of serum creatinine,urine creatinine and urine albumin were measured.PAS and Masson stainings were used to analyze the renal glomerulus morphology and interstitial fibrosis.Immunohistochemical assay and Western blotting were performed to detect the expression of CPT1A and fibrosis markers.In in vitro study,human proximal tubular epithelial HK-2 cells were divided into Ctrl group,HG group(30 mmol/L glucose treatment),and M group(30 mmol/L mannitol treatment).After treatment for 48 h,the protein levels of CPT1A and fibrosis markers were detected with Western blotting.After HK-2 cells were transfected with CPT1A overexpression plasmid and then treated with 30 mmol/L glucose,cell morphological changes and the expression of CPT1A and fibrosis markers were observed.ResultsThe mice of the DKD group had obvious glomerulus hypertrophy,mesangial expansion and renal interstitial fibrosis,increased levels of serum creatinine and urine albumin/creatinine ratio(P<0.05),decreased CPT1A expression,and elevated protein levels of fibrosis markers when compared with the mice of the Ctrl group.In in vitro experiment,HG treatment induced the morphological changes of HK-2 cells into long spindle-shape,decreased level of CPT1A,and increased levels of fibrosis markers,which were not affected by alterations of osmotic pressure.However,CPT1A overexpression significantly reversed these HG-induced morphological chang

关 键 词：肉碱棕榈酰转移酶1A 糖尿病肾病 肾间质纤维化 人近端肾小管上皮细胞 

分 类 号：R363.22[医药卫生—病理学] R587.2[医药卫生—基础医学] R692