CAJ | 학술논문

旨在探讨叉头状转录因子O1(FoxO1)对糖尿病大鼠足细胞Ⅳ型胶原和结蛋白(desmin)表达的影响.建立糖尿病大鼠模型,随机分为糖尿病+空慢病毒载体(LV-pSC-GFP)感染组(LV-NC组)、糖尿病+大鼠结构性活性FoxO1慢病毒载体(LV-FoxO1-AAA)感染组(LV-CA组)、糖尿病组(DM组),另设对照组.于慢病毒感染后的2、4、8周末,测大鼠尿白蛋白、体重、血糖、血肌酐、尿素氮,光镜和透射电镜下观察肾小球及其足细胞结构变化,实时定量PCR和Western印迹法检测大鼠肾皮质中FoxO1、Ⅳ型胶原α3链(COL4A3)、Ⅳ型胶原α5链(COL4A5)、desmin mRNA和蛋白的表达.与LV-NC、DM组相比,LV-CA组大鼠肾脏FoxO1 mRNA、蛋白表达水平明显升高(P＜0.05),尿白蛋白、血肌酐、尿素氮明显降低(2周除外)(P＜0.05),肾脏中COL4A3、COL4A5、desmin mRNA和蛋白水平也明显降低(P＜0.05),肾脏病理学变化有明显改善.通过靶向注射慢病毒载体来上调FoxO1的表达可改善糖尿病大鼠足细胞Ⅳ型胶原和desmin的异常表达.
지재탐토차두상전록인자O1(FoxO1)대당뇨병대서족세포Ⅳ형효원화결단백(desmin)표체적영향.건립당뇨병대서모형,수궤분위당뇨병+공만병독재체(LV-pSC-GFP)감염조(LV-NC조)、당뇨병+대서결구성활성FoxO1만병독재체(LV-FoxO1-AAA)감염조(LV-CA조)、당뇨병조(DM조),령설대조조.우만병독감염후적2、4、8주말,측대서뇨백단백、체중、혈당、혈기항、뇨소담,광경화투사전경하관찰신소구급기족세포결구변화,실시정량PCR화Western인적법검측대서신피질중FoxO1、Ⅳ형효원α3련(COL4A3)、Ⅳ형효원α5련(COL4A5)、desmin mRNA화단백적표체.여LV-NC、DM조상비,LV-CA조대서신장FoxO1 mRNA、단백표체수평명현승고(P＜0.05),뇨백단백、혈기항、뇨소담명현강저(2주제외)(P＜0.05),신장중COL4A3、COL4A5、desmin mRNA화단백수평야명현강저(P＜0.05),신장병이학변화유명현개선.통과파향주사만병독재체래상조FoxO1적표체가개선당뇨병대서족세포Ⅳ형효원화desmin적이상표체.
To study the effects of forkhead transcription factor O1 (FoxO1) on the expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.Streptozotocin-induced diabetic rats were divided into three groups:diabetic rats (DM group),rats transfected with blank lentiviral vectors (diabetes mellitus plus LV-pSC-GFP group,LV-NC group),and rats which were transfected with lentiviral vectors carrying constitutively active FoxO1 (diabetes mellitus plus LV-FoxO1-AAA group,LV-CA group).Rats which received an injection of diluent buffer served as normal control.At 2,4,and 8 weeks after transfection,the levels of urine albumin,blood glucose,blood urea nitrogen,and serum creatinine were measured.Realtime PCR and Western blotting were performed to measure the mRNA and protein levels of FoxO1,COL4A3,COL4A5,and desmin in the renal cortex.Moreover,light microscope and electron microscope were used to observe the structural changes in glomerulus and podocytes.Compared with LV-NC and DM group,in LV-CA group,there was a significant increase in the mRNA and protein levels of FoxO1,and a distinct decrease in the levels of urine albumin,blood urea nitrogen and serum creatinine of rats (except at the twoweek time point) (all P＜0.05),the mRNA and protein levels of COL4A3,COL4A5,and desmin were all decreased (all P＜0.05),and pathological changes in kidney were also improved.Upregulating the expression of FoxO1 by transfecting with constructed lentiviral vectors can definitely improve the abnormal expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.