CAJ | 학술논문

目的观察在高糖刺激下纤连蛋白(FN)与整合素连接激酶(ILK)在肾小管上皮细胞的表达情况,探讨糖尿病肾病小管间质纤维化的发病机制.方法以人肾小管上皮细胞株(HKC)细胞和链脲佐菌素(STZ)诱导的CD-1小鼠糖尿病动物模型为研究对象,采用Western印迹的方法检测细胞或肾组织ILK和FN蛋白的表达.通过基凶转染的方法,将含无激酶活力的人ILK基因表达质粒pCMV-kdILK转染HKC细胞,观察抑制ILK活力对高糖诱导的HKC合成FN的影响.结果 STZ注射后4周,CD-1小鼠血糖水平显著高于对照组[(20.3±2.7)mmol/L比(6.1±1.4)mmol/L,P＜0.01],同时,肾组织ILK和FN的表达量亦显著高于对照组,且ILK与FN表达量呈正相关(P＜0.01).细胞培养实验证实,高糖能够刺激HKC上调ILK和FN蛋白的表达,并且呈时间和剂量依赖性.HKC细胞转染pCMV-kdILK质粒以抑制ILK的活力,能够显著地拮抗高糖刺激HKC表达FN的作用.结论高糖通过上调ILK蛋白增加肾小管表达并合成FN,抑制ILK能够拮抗高糖刺激HKC合成FN的作用.
목적 관찰재고당자격하섬련단백(FN)여정합소련접격매(ILK)재신소관상피세포적표체정황,탐토당뇨병신병소관간질섬유화적발병궤제.방법 이인신소관상피세포주(HKC)세포화련뇨좌균소(STZ)유도적CD-1소서당뇨병동물모형위연구대상,채용Western인적적방법검측세포혹신조직ILK화FN단백적표체.통과기흉전염적방법,장함무격매활력적인ILK기인표체질립pCMV-kdILK전염HKC세포,관찰억제ILK활력대고당유도적HKC합성FN적영향.결과 STZ주사후4주,CD-1소서혈당수평현저고우대조조[(20.3±2.7)mmol/L비(6.1±1.4)mmol/L,P＜0.01],동시,신조직ILK화FN적표체량역현저고우대조조,차ILK여FN표체량정정상관(P＜0.01).세포배양실험증실,고당능구자격HKC상조ILK화FN단백적표체,병차정시간화제량의뢰성.HKC세포전염pCMV-kdILK질립이억제ILK적활력,능구현저지길항고당자격HKC표체FN적작용.결론 고당통과상조ILK단백증가신소관표체병합성FN,억제ILK능구길항고당자격HKC합성FN적작용.
Objective To investigate the relationship between high-glucose-induced fibronectin(FN) expression and up-regulation of integrin-linked kinase(ILK) in human kidney tubular epithelial cells (HKC) and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin (STZ)-indueed diabetic model of CD-1 mice were enrolled in this study.Western blot was used to detect the expression of FN and ILK.The kinase dead ILK plasmid (pCMV-kdlLK) were transferred to HKC. Results Four weeks after injection of STZ,CD-1 mice had higher blood glucose level as compared to the control [(20.3±2.7) mmol/L vs (6.1±1.4) mmol/L,P＜0.01].Meanwhile,expression of FN and ILK was significantly increased in diabetic mice as compared to the control (P＜0.01).There was positive correlation between the expression of FN and ILK (r=0.899,P＜0.01).High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner.Blockage of ILK activation by pCMV-kdILK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression.Blockage of ILK activation abrogates this effect.