CAJ | 학술논문

目的探讨雷帕霉素对嘌呤霉素氨基核苷(PAN)肾病模型小鼠肾脏病变和肾小球VEGF及受体(VEGFR)表达的影响.方法 24只BALB/c小鼠随机分为PAN 组(单次尾静脉注射PAN造模,n=8)、雷帕霉素干预组(单次尾静脉注射PAN造模+雷帕霉素干预,n=8)和对照组(单次尾静脉注射PBS,n=8).各组动物留取24 h尿液,BCA蛋白定量试剂盒测定24 h尿蛋白排泄量.于造模后第10天处死动物取肾脏制备肾组织标本,透射电子显微镜观察各组肾小球足突结构改变;分别采用Real-time PCR、Western b1otting和免疫组织化学方法检测各组肾组织VEGF、VEGFRI、VEGFR2 mRNA以及VEGF、VEGFR2蛋白表达.结景24 h尿蛋白排泄量为PAN组>雷帕霉素干预组>对照组,组间差异均有统计学意义(P<0.05).电子显微镜观察发现PAN 组肾小球上皮细胞足突广泛融合.各组肾组织VEGF、VEGFRI、VEGFR2 mRNA及蛋白表达为PAN组>雷帕霉素干预组>对照组(P<0.05),且各组VEGF、VEGFRI、VEGFR2 mRNA及VEGF蛋白表达与24 h尿蛋白排泄量呈正相关(P<0.05).结论雷帕霉素能减轻PAN肾病小鼠蛋白尿的程度,作用机制可能与下调肾小球VEGF及其受体基因表达有关.
목적 탐토뢰파매소대표령매소안기핵감(PAN)신병모형소서신장병변화신소구VEGF급수체(VEGFR)표체적영향.방법 24지BALB/c소서수궤분위PAN 조(단차미정맥주사PAN조모,n=8)、뢰파매소간예조(단차미정맥주사PAN조모+뢰파매소간예,n=8)화대조조(단차미정맥주사PBS,n=8).각조동물류취24 h뇨액,BCA단백정량시제합측정24 h뇨단백배설량.우조모후제10천처사동물취신장제비신조직표본,투사전자현미경관찰각조신소구족돌결구개변;분별채용Real-time PCR、Western b1otting화면역조직화학방법검측각조신조직VEGF、VEGFRI、VEGFR2 mRNA이급VEGF、VEGFR2단백표체.결경24 h뇨단백배설량위PAN조>뢰파매소간예조>대조조,조간차이균유통계학의의(P<0.05).전자현미경관찰발현PAN 조신소구상피세포족돌엄범융합.각조신조직VEGF、VEGFRI、VEGFR2 mRNA급단백표체위PAN조>뢰파매소간예조>대조조(P<0.05),차각조VEGF、VEGFRI、VEGFR2 mRNA급VEGF단백표체여24 h뇨단백배설량정정상관(P<0.05).결론 뢰파매소능감경PAN신병소서단백뇨적정도,작용궤제가능여하조신소구VEGF급기수체기인표체유관.
Objective To investigate the effects of Rapamycin on nephropathy and expression of vascular endothelial growth factors (VEGF) and VEGF receptors ( VEGFR) of glomeruli in puromycin amino nuclear glucoside ( PAN) nephritic mice models. Methods Twenty-four BALB/c mice were randomly divided into PAN group ( model establishment by single injection of PAN via tail vein, n = 8), Rapamycin intervention group (model establishment by single injection of PAN via tail vein + Rapamycin intervention, n = 8) and control group ( single injection of PBS via tail vein, n = 8). Twenty-four-hour urine was obtained from each group, and urinary protein excretion was determined by BCA protein assay. Mice were sacrificed 10 days after model establishment, and renal tissue samples were prepared. The structural changes of foot process of glomeruli in each group was observed by transmission electron microscope, and the expression of VEGF, VEGFR1 and VEGFR2 mRNA and VEGF and VEGFR2 protein was detected by Real-time PCR, Western blotting and immunohistochemistry. Results There were significant differences in 24-h urinary protein excretion among groups (P <0. 05), with PAN group > Rapamycin intervention group > control group. Foot process infusion in glomeruli was observed by electron microscopy. The expression of VEGF, VEGFR1 and VEGFR2 mRNA and protein in PAN group was the highest, and that in control group was the lowest, with significant differences among groups (P<0. 05). The expression of VEGF, VEGFR 1 and VEGFR2 mRNA and protein was significantly correlated with 24-h urinary protein excretion in each group (P <0.05). Conclusion Rapamycin can reduce the proteinuria of PAN nephritic mice, and the mechanism may be related to the down-regulation of expression of VEGF and VEGFR of glomeruli.