摘要
目的:分析肾间质纤维化(RIF)早期的蛋白质组表达变化情况,为探讨干预RIF的新靶点奠定基础。方法:(1)48只雄性Wistar大鼠,随机分为正常对照(control)组(6只)、假手术(sham)组(6只)和单侧输尿梗阻(UUO)模型组(36只)。UUO组行右侧输尿管结扎,并分别于术后第3、7、11、14、21和28天各处死6只,取肾脏标本行HE染色和Masson染色,光镜下观察病理变化;control组不作任何处理,sham组开腹后只分离右输尿管但不结扎,两组均于第28天取肾脏标本作病理观察;(2)取UUO术后第7天大鼠梗阻侧和健侧肾组织的蛋白质提取液行双向凝胶电泳,两侧对照找出差异蛋白质,选取部分差异蛋白点进行质谱鉴定;(3)Western blot检测UUO大鼠肾脏组织膜联蛋白A5(annexin A5)表达的动态变化。结果:(1)UUO各组梗阻肾脏逐渐出现肾小管扩张、炎症细胞浸润、肾小管上皮细胞萎缩、间质胶原纤维沉积等变化,其中,肾梗阻后第7天开始出现间质纤维化;(2)与健侧肾比较,UUO术后第7天的梗阻侧肾出现≥1.5倍的差异蛋白质166个,其中上调81个,下调85个,质谱成功鉴定出19种差异蛋白质;(3)Western blot显示,各时段UUO组大鼠梗阻侧肾组织中annexin A5表达均明显高于sham组(P<0.05),其中以梗阻第3天组的表达量最高,随后逐渐下降,但仍高于sham组。结论:(1)RIF早期存在大量表达差异的蛋白质;(2)RIF的annexin A5表达显著增加,可能与某些病理变化存在密切关系。
AIM:To explore the novel intervention targets of renal interstitial fibrosis(RIF)by the analysis of renal proteome expression in the early stage of RIF.METHODS:(1)Male Wistar rats(n=48)were randomly divided into normal control group(control group),sham operation group(sham group)and unilateral ureteral obstruction(UUO)model group(UUO group).The right ureters were ligated in UUO group,and the kidney specimens of the animals were taken for pathological analysis with HE staining and Masson staining on the 3rd,7th,11th,14th,21st and 28th day after operation.No treatment was given to the rats in control group,and the right ureter was separated but not ligated in sham group.The same pathological analysis as UUO group was performed in control group and sham group on the 28th day.(2)Two-dimensional gel electrophoresis(2-DE)of the kidney tissue proteins from 3 UUO rats was performed 7 days after operation for the discovery of differentially-expressed proteins between the 2 sides of the kidneys,and then some of differential proteins were selected for mass spectrometry(MS).(3)The expression changes of annexin A5 in the kidney tissue of UUO rats were observed by Western blot.RESULTS:(1)Renal interstitial fibrosis developed gradually after ureteral obstruction,with its initial appearance on the 7th day after operation.(2)The results of 2-DE analysis of the 7th day after UUO operation showed that there were 166 differential proteins with more than 1.5 times of difference between the 2 sides of the kidneys,in which 81 proteins were up-regulated and 85 were down-regulated in the obstructive kidney compared with the non-obstructive kidney,and 19 differential proteins were successfully identified.(3)The results of Western blot showed that the expression of annexin A5 in the obstructed kidney of UUO rats was significantly higher than that in sham group(P<0.05),with a peak on the 3rd day of obstruction.CONCLUSION:There are a large number of differentiallyexpressed proteins in the early stage of RIF.The expression of annexin A5 is si
作者
李肖肖
陆大祥
王华东
戚仁斌
张珂珂
王达安
LI Xiao-xiao;LU Da-xiang;WANG Hua-dong;QI Ren-bin;ZHANG Ke-ke;WANG Da-an(Department of Pathophysiology&Key Laboratory of State Administration of Traditional Chinese Medicine,College of Ba-sic Medicine,Jinan University,Guangzhou 510632,China;Nursing Division,The Second People's Hospital of Foshan,Foshan 528500,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第3期504-511,共8页
Chinese Journal of Pathophysiology
基金
广东省科技计划项目(No.2014A020212217)。
