中国中西医结合肾病杂志
中國中西醫結閤腎病雜誌
중국중서의결합신병잡지
CHINESE JOURNAL OF INTEGRATED TRADITIONAL AND WESTERN NEPHROLOGY
2014年
7期
578-582
,共5页
艾娜%谢席胜%龙琼先%王彦江%夏梦迪
艾娜%謝席勝%龍瓊先%王彥江%夏夢迪
애나%사석성%룡경선%왕언강%하몽적
链脲佐菌素%血小板反应蛋白-1%糖尿病肾损害%蛋白尿
鏈脲佐菌素%血小闆反應蛋白-1%糖尿病腎損害%蛋白尿
련뇨좌균소%혈소판반응단백-1%당뇨병신손해%단백뇨
StrePtozotocin( STZ)%ThrombosPondin-1%Diabetic nePhroPathy%Proteinuria
目的：观察血小板反应蛋白-1（thrombospondin-1，TSP-1）在链脲佐菌素诱导的糖尿病（diabetes mellitus， DM）大鼠模型肾脏中的表达，初步探讨TSP-1在糖尿病肾病大鼠发病机制中的作用及与糖尿病肾损害的关系。方法：成年雄性SD大鼠80只，随机分为正常对照组（ n＝15）和实验组（ n＝65）。采用STZ 70 mg/kg单次腹腔注射建立DM大鼠模型，按照成模标准去除未成模大鼠。检测各组大鼠空腹血糖（ fasting blood glucose，FBG）、尿蛋白及肾功能；分别于2，4，8周处死各组大鼠，通过HE染色观察各组大鼠肾脏的病理改变，免疫荧光观察TSP-1在各组大鼠肾脏的分布；PCR检测肾组织TSP-1和转录生长因子β1（ transforming growth factor，TGF-β1）基因表达水平；采用酶联免疫吸附法（ ELISA）检测肾组织活性TGF-β1水平；Western杂交检测TSP-1的动态变化。分别对尿蛋白、肾功能、肾脏病理损害、活性TGF-β1与TSP-1蛋白表达的相关性进行分析。结果：HE染色可见DM组大鼠肾小球系膜基质增生，肾小球可见分叶现象、K-W结节等典型的糖尿病肾病损伤特点。与正常对照组相比，DM组大鼠的FBG、尿蛋白、肌酐、尿素氮水平均显著增加（ P﹤0．05），TSP-1在DM组大鼠主要分布于肾小管、肾小球系膜细胞，其蛋白表达在2周时即明显增加，一直持续到第8周。DM 组大鼠肾脏TSP-1的积分光密度（ integrated optical density，IOD）与尿蛋白、肾脏病理损害、肌酐、尿素氮水平、活性TGF-β1浓度均呈正相关。结论：持续高血糖状态可以诱导大鼠肾脏病理损害，引起尿蛋白升高、肾功能异常；高血糖可诱导大鼠肾脏TSP-1蛋白高表达。TSP-1蛋白的高表达可能通过激活TGF-β1信号通路参与DM大鼠蛋白尿的产生及肾损害的发生发展。
目的：觀察血小闆反應蛋白-1（thrombospondin-1，TSP-1）在鏈脲佐菌素誘導的糖尿病（diabetes mellitus， DM）大鼠模型腎髒中的錶達，初步探討TSP-1在糖尿病腎病大鼠髮病機製中的作用及與糖尿病腎損害的關繫。方法：成年雄性SD大鼠80隻，隨機分為正常對照組（ n＝15）和實驗組（ n＝65）。採用STZ 70 mg/kg單次腹腔註射建立DM大鼠模型，按照成模標準去除未成模大鼠。檢測各組大鼠空腹血糖（ fasting blood glucose，FBG）、尿蛋白及腎功能；分彆于2，4，8週處死各組大鼠，通過HE染色觀察各組大鼠腎髒的病理改變，免疫熒光觀察TSP-1在各組大鼠腎髒的分佈；PCR檢測腎組織TSP-1和轉錄生長因子β1（ transforming growth factor，TGF-β1）基因錶達水平；採用酶聯免疫吸附法（ ELISA）檢測腎組織活性TGF-β1水平；Western雜交檢測TSP-1的動態變化。分彆對尿蛋白、腎功能、腎髒病理損害、活性TGF-β1與TSP-1蛋白錶達的相關性進行分析。結果：HE染色可見DM組大鼠腎小毬繫膜基質增生，腎小毬可見分葉現象、K-W結節等典型的糖尿病腎病損傷特點。與正常對照組相比，DM組大鼠的FBG、尿蛋白、肌酐、尿素氮水平均顯著增加（ P﹤0．05），TSP-1在DM組大鼠主要分佈于腎小管、腎小毬繫膜細胞，其蛋白錶達在2週時即明顯增加，一直持續到第8週。DM 組大鼠腎髒TSP-1的積分光密度（ integrated optical density，IOD）與尿蛋白、腎髒病理損害、肌酐、尿素氮水平、活性TGF-β1濃度均呈正相關。結論：持續高血糖狀態可以誘導大鼠腎髒病理損害，引起尿蛋白升高、腎功能異常；高血糖可誘導大鼠腎髒TSP-1蛋白高錶達。TSP-1蛋白的高錶達可能通過激活TGF-β1信號通路參與DM大鼠蛋白尿的產生及腎損害的髮生髮展。
목적：관찰혈소판반응단백-1（thrombospondin-1，TSP-1）재련뇨좌균소유도적당뇨병（diabetes mellitus， DM）대서모형신장중적표체，초보탐토TSP-1재당뇨병신병대서발병궤제중적작용급여당뇨병신손해적관계。방법：성년웅성SD대서80지，수궤분위정상대조조（ n＝15）화실험조（ n＝65）。채용STZ 70 mg/kg단차복강주사건립DM대서모형，안조성모표준거제미성모대서。검측각조대서공복혈당（ fasting blood glucose，FBG）、뇨단백급신공능；분별우2，4，8주처사각조대서，통과HE염색관찰각조대서신장적병리개변，면역형광관찰TSP-1재각조대서신장적분포；PCR검측신조직TSP-1화전록생장인자β1（ transforming growth factor，TGF-β1）기인표체수평；채용매련면역흡부법（ ELISA）검측신조직활성TGF-β1수평；Western잡교검측TSP-1적동태변화。분별대뇨단백、신공능、신장병리손해、활성TGF-β1여TSP-1단백표체적상관성진행분석。결과：HE염색가견DM조대서신소구계막기질증생，신소구가견분협현상、K-W결절등전형적당뇨병신병손상특점。여정상대조조상비，DM조대서적FBG、뇨단백、기항、뇨소담수평균현저증가（ P﹤0．05），TSP-1재DM조대서주요분포우신소관、신소구계막세포，기단백표체재2주시즉명현증가，일직지속도제8주。DM 조대서신장TSP-1적적분광밀도（ integrated optical density，IOD）여뇨단백、신장병리손해、기항、뇨소담수평、활성TGF-β1농도균정정상관。결론：지속고혈당상태가이유도대서신장병리손해，인기뇨단백승고、신공능이상；고혈당가유도대서신장TSP-1단백고표체。TSP-1단백적고표체가능통과격활TGF-β1신호통로삼여DM대서단백뇨적산생급신손해적발생발전。
Objective:Study the expression of thrombos pondin-1(TSP-1)in the kidney of diabetes(DM)rat model in-duced by streptozotocin( STZ)and discussion the role in the pathogenesis of diabetic nephropathy and the relationship with the DM kidney injury. Methods:A total of 80 male Sprague-Dawley rats were randomly divided into normal control group(n=15)and ex-perimental group(n=65). ExPerimental guoup were treated with an intraperitoneal injection dose(70 mg/kg)of STZ to establish the model of DM. After the injection of STZ,the fasting blood glucose of rats in the experimental group does not meet the standard of model were excluded. The level of the fasting blood glucose( FBG),urine protein( UP)and renal function were monitored in each group;rats in each group were sacrificed at 2,4 and 8 weeks,respectively,use HE staining observe the pathological changes of kid-ney;the expression of TSP-1 was measured by immunohistochemical in the rat kidney;mRNA of TSP-1 and TGF-β1 were reverse transcribed and quantified by PCR;enzyme linked immune sorbent assay( ELISA)were performed to examine the protein expression levels of active TGF-β1 in kidney. Western Blot to detect the expression of TSP-1 protein in rat kidney. the correlation between u-rine protein,pathological damage,renal function and TSP-1 protein expression were analyzed. Results:HE staining showed mesang-ial and matrix proliferative in DM rats,some typical damage characteristics of diabetic nephropathy can be found in glomerular include leaf Phenomenon,K-W nodules. Compared with normal control group,FBG,UP,creatinine and urea nitrogen was significantly in-creased in DM rats(P﹤0. 05). TSP-1 is mainly distributed in DM rats in the renal tubules,mesangial cells,its expression was markedly increased at two weeks time,until the eighth week. The integrated optical density( IOD)of TSP-1 were positively correla-ted with urinary protein,renal pathological damage,creatinine,urea nitrogen and concentration of active TGF-β1 . Conclusion:Sus-tained hyperglycemia can induce renal pathological damage,causing UP increased and renal dysfunction;hyperglycemia can induce the expression of TSP-1 increase in renal. The high expression of TSP-1 protein may be involve the appear and development of pro-teinuria and renal damage in DM rata through active the TGF-β1 signal axonal.
