CAJ | 학술논문

目的探讨静脉注射小鼠脂联素基因重组慢病毒对糖尿病肾病(DN)模型小鼠的肾脏保护作用及其机制.方法 40只C57BL/6小鼠按数字随机法分为正常对照组(NC组)、糖尿病肾病组(DN组)、阴性对照病毒(Lenti-IRES-EGFP)治疗组(DL组)、脂联素基因重组慢病毒( Lenti-Acdc-IRES-EGFP)治疗组(DA组),每组10只.应用链脲菌素(SFZ)结合高脂饮食诱导建立DN模型.重组慢病毒注射8周后,检测各组小鼠血浆脂联素水平、肾功能、尿白蛋白排泄率、肾组织病理改变.用免疫组织化学法原位检测肾组织增殖细胞核抗原( PCNA)表达.用Western印迹检测肾组织腺苷酸活化蛋白激酶α(AMPKα)、哺乳动物雷帕霉素靶蛋白( mTOR)水平.结果成功构建脂联素超表达DN动物模型.与NC组比较,第12周末DA组小鼠肾质量指数(肾质量/体质量,KWI)、平均肾小球体积(MGV)、系膜面积比(FMA)、24h尿蛋白(UTP)均明显升高(P＜0.05),但低于DN组、DL组(P＜0.05).DA组肾组织PCNA阳性细胞数显著低于DN组、DL组(P＜0.01).与DN组、DL组相比,DA组小鼠肾组织AMPKα磷酸化水平显著升高(P＜0.01),mTOR磷酸化水平降低(P＜0.01).结论脂联素通过激活AMPK信号通路,抑制mTOR信号活化,进而抑制早期DN时肾组织异常增殖.
목적 탐토정맥주사소서지련소기인중조만병독대당뇨병신병(DN)모형소서적신장보호작용급기궤제.방법 40지C57BL/6소서안수자수궤법분위정상대조조(NC조)、당뇨병신병조(DN조)、음성대조병독(Lenti-IRES-EGFP)치료조(DL조)、지련소기인중조만병독( Lenti-Acdc-IRES-EGFP)치료조(DA조),매조10지.응용련뇨균소(SFZ)결합고지음식유도건립DN모형.중조만병독주사8주후,검측각조소서혈장지련소수평、신공능、뇨백단백배설솔、신조직병리개변.용면역조직화학법원위검측신조직증식세포핵항원( PCNA)표체.용Western인적검측신조직선감산활화단백격매α(AMPKα)、포유동물뢰파매소파단백( mTOR)수평.결과 성공구건지련소초표체DN동물모형.여NC조비교,제12주말DA조소서신질량지수(신질량/체질량,KWI)、평균신소구체적(MGV)、계막면적비(FMA)、24h뇨단백(UTP)균명현승고(P＜0.05),단저우DN조、DL조(P＜0.05).DA조신조직PCNA양성세포수현저저우DN조、DL조(P＜0.01).여DN조、DL조상비,DA조소서신조직AMPKα린산화수평현저승고(P＜0.01),mTOR린산화수평강저(P＜0.01).결론 지련소통과격활AMPK신호통로,억제mTOR신호활화,진이억제조기DN시신조직이상증식.
Objective To investigate the renal protective effect of recombinant lentivirus encoding adiponectin gene on streptozocin-induced early diabetic nephropathy (DN) mice,and to explore its potential mechanism. Methods Forty C57BL/6 mice were randomly divided into normal control group (NC group,n=10),diabetic nephropathy group (DN group,n=10),LentiIRES-EGFP treatment group (DL group,n=10) and Lenti-Acdc-IRES-EGFP treatment group (DA group,n=10).After 8 weeks of recombinant lentivirus injection,kidney to body weight ratio (KW/BW),mean glomerular volume (MGV),fractional mesangial area (FMA),24 h urinary protein excretion (UTP),Scr,BUN,serum albumin and adiponectin were measured.Renal pathological changes were evaluated by electron microscopy.Proliferation of glomendar and tubulointerstitial cells was assessed by immunohistochemistry using PCNA antibody.The phosphorylation of AMP-activated protein kinase (AMPK) and mammalian target of rapamycin protein (mTOR) were detected by Western blotting. Results Adiponectin was successfully over-expressed in STZ-induced DN mice after lentivirus injection.KW/BW,MGV,FMA and UTP were significantly decreased in DA group as compared to DN group and DL group (P＜0.05),but were increased as compared to NC group (P＜0.05).DA group animals had significantly fewer PCNA-positive cells than DN group and DL group (P＜0.01).DA group mice had higher p-AMPK level and lower p-mTOR level as compared to DN group and DL group (P＜0.01). Conclusion Over-expression of adiponectin has beneficial effect on early DN and attenuates aberrant proliferation of renal cells via AMPKmTOR pathway.