CAJ | 학술논문

目的探讨胰岛移植改善糖尿病肾病(DN)大鼠足细胞损害的作用及其机制.方法采用腹腔注射单剂链脲菌素建立SD大鼠DN模型(简称:建模).实验分正常对照组、DN组和胰岛移植组.胰岛移植组在建模后8周行右侧肾背膜下胰岛移植.于建模第12周(移植后4周),测定各组大鼠尿微量白蛋白与尿肌酐比值,监测各组大鼠血糖水平;收集各组大鼠肾脏行病理组织检查和免疫荧光检查及肾脏电镜检测;采用免疫组织化学法观察各组肾小球中突触极蛋白(synaptopodin)和转化生长因子β1(TGF-β1)蛋白的表达.结果建模后8周(移植前),DN组和胰岛移植组大鼠血糖水平分别为(24.62±3.31) mmol/L和(25.23±2.61) mmol/L,均较正常对照组显著升高(P＜0.05);胰岛移植组大鼠在移植后平均3.3d,血糖下降至正常,并均可维持正常;而此时DN组大鼠的血糖水平未出现明显变化,两组差异有统计学意义(P＜0.05).建模第8周(移植前),DN组和胰岛移植组24 h尿蛋白水平分别为(46.82±5.87)mg和(51.26±4.69)mg,均显著高于正常组的(8.07±1.17)mg(P＜0.05).建模第12周(移植后4周),胰岛移植组尿微量白蛋白与尿肌酐比值水平为(0.33±0.04) mg/mmol,显著低于DN组的(2.36±0.73)mg/mmol,差异有统计学意义(P＜0.001).HE染色和免疫荧光染色显示,胰岛团周围有新生血管,胰岛素分泌旺盛.电镜下可见,DN组足细胞局部足突融合,基底膜节段增厚;胰岛移植组足突排列整齐,基底膜结构清晰,无增厚.DN组和胰岛移植组肾小球内synaptopodin蛋白表达较正常对照组明显减弱(P＜0.05);胰岛移植组synaptopodin 蛋白表达较DN组显著增强(P＜0.05).DN组和胰岛移植组TGF-β1蛋白表达较正常对照组显著升高(P＜0.05);DN组TGF-β1蛋白表达较胰岛移植组显著升高(P＜0.05).结论胰岛移植可以通过抑制TGF-β1通路,改善DN大鼠足细胞损伤,减少甚至逆转蛋白尿的产生.
목적 탐토이도이식개선당뇨병신병(DN)대서족세포손해적작용급기궤제.방법 채용복강주사단제련뇨균소건립SD대서DN모형(간칭:건모).실험분정상대조조、DN조화이도이식조.이도이식조재건모후8주행우측신배막하이도이식.우건모제12주(이식후4주),측정각조대서뇨미량백단백여뇨기항비치,감측각조대서혈당수평;수집각조대서신장행병리조직검사화면역형광검사급신장전경검측;채용면역조직화학법관찰각조신소구중돌촉겁단백(synaptopodin)화전화생장인자β1(TGF-β1)단백적표체.결과 건모후8주(이식전),DN조화이도이식조대서혈당수평분별위(24.62±3.31) mmol/L화(25.23±2.61) mmol/L,균교정상대조조현저승고(P＜0.05);이도이식조대서재이식후평균3.3d,혈당하강지정상,병균가유지정상;이차시DN조대서적혈당수평미출현명현변화,량조차이유통계학의의(P＜0.05).건모제8주(이식전),DN조화이도이식조24 h뇨단백수평분별위(46.82±5.87)mg화(51.26±4.69)mg,균현저고우정상조적(8.07±1.17)mg(P＜0.05).건모제12주(이식후4주),이도이식조뇨미량백단백여뇨기항비치수평위(0.33±0.04) mg/mmol,현저저우DN조적(2.36±0.73)mg/mmol,차이유통계학의의(P＜0.001).HE염색화면역형광염색현시,이도단주위유신생혈관,이도소분비왕성.전경하가견,DN조족세포국부족돌융합,기저막절단증후;이도이식조족돌배렬정제,기저막결구청석,무증후.DN조화이도이식조신소구내synaptopodin단백표체교정상대조조명현감약(P＜0.05);이도이식조synaptopodin 단백표체교DN조현저증강(P＜0.05).DN조화이도이식조TGF-β1단백표체교정상대조조현저승고(P＜0.05);DN조TGF-β1단백표체교이도이식조현저승고(P＜0.05).결론 이도이식가이통과억제TGF-β1통로,개선DN대서족세포손상,감소심지역전단백뇨적산생.
Objective To investigate the mechanism of islet transplantation lessening renal damage of diabetic nephropathy (DN) rat model.Method Rat DN model was established by intraperitoneal injection of a single-dose streptozotocir.The rats were divided into normal control group, DN group and islet transplant group.Islet transplantation was taken on the right renal capsule in transplantation group at 8th week after the modeling.At week 12 after the modeling, the urinary protein, urinary creatinine and blood glucose in each group were determined.The kidneys were collected, and transplant islet HE staining, immunofluorescence, kidney electron microscopy were done.Synaptopodin and transforming growth factor-β1 (TGFβ1) protein expression was observed in renal tissues of each group by immunohistochemical staining.Result The urine protein and urinary creatinine ratio in islet transplant group was significantly lower than in DN group (P＜0.001).Blood glucose level in islet transplant group had no significant difference (P＞0.05) with the control group, but was significantly lower than in DN group (P ＜ 0.05).Islet cells by HE staining and immunofluorescence staining showed new blood vessels around the islets, and the insulin secretion was exuberant.Under an electron microscope, there was local fusion of podocyte foot processes, and segmental thickening of the basement membrane in DN group;in islet transplant group, the foot processes of podocytes were neat, basement membrane structure was clear and had no thickening.Synaptopodin protein expression was significantly decreased in the glomeruli of DN group and islet transplant group as compared with the control group (P＜0.05), and that in islet transplant group was significantly enhanced (P＜0.05) as compared with DN group.The expression of TGFβ1 in DN group and islet transplant group was significantly increased (P＜0.05) as compared with control group, and that in DN group was significantly higher (P＜0.05) than in islet transplant group.Conclusion Islet transplantation can inhibit TGFβ1 pathway, improve DN podocyte injury in rats, and alleviate or even reverse proteinuria.