中华肾病研究电子杂志
中華腎病研究電子雜誌
중화신병연구전자잡지
2015年
1期
37-42
,共6页
单侧输尿管梗阻%血小板反应蛋白-1%CD47%肾周毛细血管病变
單側輸尿管梗阻%血小闆反應蛋白-1%CD47%腎週毛細血管病變
단측수뇨관경조%혈소판반응단백-1%CD47%신주모세혈관병변
Unilateral ureteral obstruction%Thrombospondin-1%CD47%Peritubular capillary lesion
目的：观察血小板反应蛋白-1（TSP-1）及其受体 CD47在单侧输尿管梗阻（UUO）大鼠模型中的表达，探讨 TSP-1-CD47在肾周毛细血管（PTC）病变中的作用。方法将60只 SD 大鼠随机分为2组：UUO 组45只和假手术组（SOR）1 5只。UUO 组随机选取 1 5只分别于术后3 d、7 d、14 d 处死；SOR 组随机选取 5只于相同时间点处死。检测24 h 尿蛋白定量，采用全自动生化分析仪检测各组大鼠的肾功能，通过 Masson 染色观察肾间质病理改变；免疫荧光双染色观察 TSP-1及其受体 CD47在 UUO 模型中是否存在共表达。Western 印迹检测毛细血管病变指标 CD34和血管内皮生长因子（VEGF）及 TSP-1的表达；多组间均数比较使用方差分析；TSP-1与 CD34、VEGF 的相关性进行Pearson 相关分析。结果与 SOR 组相比，UUO 各组大鼠血清肌酐、尿素氮、24 h 尿蛋白定量随梗阻时间延长无明显变化，但肾间质纤维化逐渐加重，肾小管损伤评分增加。免疫荧光双染色显示 TSP-1及其受体 CD47在肾小管间质存在共表达。Western 印迹检测显示随 UUO 时间延长，TSP-1蛋白水平逐渐增加，而 CD34及 VEGF 蛋白表达随梗阻时间延长逐渐下降。相关分析显示 TSP-1蛋白水平与CD34及 VEGF 的表达呈负相关（r ＝－0．931，P ＜0．01；r ＝－0．953，P ＜0．01）。结论TSP-1及其受体 CD47在 UUO 大鼠模型肾间质存在共表达，且与 PTC 呈负相关。TSP-1可能通过 CD47参与 UUO肾脏 PTC 病变的发生发展。
目的：觀察血小闆反應蛋白-1（TSP-1）及其受體 CD47在單側輸尿管梗阻（UUO）大鼠模型中的錶達，探討 TSP-1-CD47在腎週毛細血管（PTC）病變中的作用。方法將60隻 SD 大鼠隨機分為2組：UUO 組45隻和假手術組（SOR）1 5隻。UUO 組隨機選取 1 5隻分彆于術後3 d、7 d、14 d 處死；SOR 組隨機選取 5隻于相同時間點處死。檢測24 h 尿蛋白定量，採用全自動生化分析儀檢測各組大鼠的腎功能，通過 Masson 染色觀察腎間質病理改變；免疫熒光雙染色觀察 TSP-1及其受體 CD47在 UUO 模型中是否存在共錶達。Western 印跡檢測毛細血管病變指標 CD34和血管內皮生長因子（VEGF）及 TSP-1的錶達；多組間均數比較使用方差分析；TSP-1與 CD34、VEGF 的相關性進行Pearson 相關分析。結果與 SOR 組相比，UUO 各組大鼠血清肌酐、尿素氮、24 h 尿蛋白定量隨梗阻時間延長無明顯變化，但腎間質纖維化逐漸加重，腎小管損傷評分增加。免疫熒光雙染色顯示 TSP-1及其受體 CD47在腎小管間質存在共錶達。Western 印跡檢測顯示隨 UUO 時間延長，TSP-1蛋白水平逐漸增加，而 CD34及 VEGF 蛋白錶達隨梗阻時間延長逐漸下降。相關分析顯示 TSP-1蛋白水平與CD34及 VEGF 的錶達呈負相關（r ＝－0．931，P ＜0．01；r ＝－0．953，P ＜0．01）。結論TSP-1及其受體 CD47在 UUO 大鼠模型腎間質存在共錶達，且與 PTC 呈負相關。TSP-1可能通過 CD47參與 UUO腎髒 PTC 病變的髮生髮展。
목적：관찰혈소판반응단백-1（TSP-1）급기수체 CD47재단측수뇨관경조（UUO）대서모형중적표체，탐토 TSP-1-CD47재신주모세혈관（PTC）병변중적작용。방법장60지 SD 대서수궤분위2조：UUO 조45지화가수술조（SOR）1 5지。UUO 조수궤선취 1 5지분별우술후3 d、7 d、14 d 처사；SOR 조수궤선취 5지우상동시간점처사。검측24 h 뇨단백정량，채용전자동생화분석의검측각조대서적신공능，통과 Masson 염색관찰신간질병리개변；면역형광쌍염색관찰 TSP-1급기수체 CD47재 UUO 모형중시부존재공표체。Western 인적검측모세혈관병변지표 CD34화혈관내피생장인자（VEGF）급 TSP-1적표체；다조간균수비교사용방차분석；TSP-1여 CD34、VEGF 적상관성진행Pearson 상관분석。결과여 SOR 조상비，UUO 각조대서혈청기항、뇨소담、24 h 뇨단백정량수경조시간연장무명현변화，단신간질섬유화축점가중，신소관손상평분증가。면역형광쌍염색현시 TSP-1급기수체 CD47재신소관간질존재공표체。Western 인적검측현시수 UUO 시간연장，TSP-1단백수평축점증가，이 CD34급 VEGF 단백표체수경조시간연장축점하강。상관분석현시 TSP-1단백수평여CD34급 VEGF 적표체정부상관（r ＝－0．931，P ＜0．01；r ＝－0．953，P ＜0．01）。결론TSP-1급기수체 CD47재 UUO 대서모형신간질존재공표체，차여 PTC 정부상관。TSP-1가능통과 CD47삼여 UUO신장 PTC 병변적발생발전。
Objective To observe the expression of TSP-1 and its receptor CD47 in unilateral ureteral obstruction (UUO)rats,and to investigate the role of TSP-1 -CD47 in lesions of peritubular capillary (PTC).Methods A total of 60 Sprague-Dawley rats were randomly divided into:sham-operation group (SOR)(n =1 5 )and UUO group (n =45 ).UUO group sacrifice timing consisted of three time points:3 day,7 day,1 4 day,with 1 5 animals for each time point.Detection of 24h urinary protein and renal function was made by automatic biochemistry analyzer,and interstitial pathological changes were shown by Masson staining.The expression of TSP-1 and CD47 was measured by immunohistochemical and immunofluorescence double staining.Western blot was used to detect the expression of CD34,VEGF,TSP-1 protein in rat kidneys.The correlations among CD34,VEGF,and TSP-1 protein expression were analyzed with Pearson method.Results Compared with the sham-operation group,the levels of serum creatinine (Scr),blood urea nitrogen (BUN),and 24h urinary protein did not change significantly in UUO group,but features of renal tubulointerstitial fibrosis were obvious,and the score of renal interstitial lesion was significantly higher in UUO group.Immunofluorescence double staining showed that TSP-1 and CD47 were coexpressed in UUO renal tubules.Western blot confirmed that TSP-1 protein level was significantly higher in UUO group,while the expressions of CD34 and VEGF protein were decreased.Correlation analysis showed that TSP-1 protein was negatively associated with the expression of CD34 and VEGF(r =-0.931 ,P <0.01 ;r =-0.953,P <0.01 ).Conclusion TSP-1 and its receptor CD47 were coexpressed in UUO rats, and negatively associated with PTC.TSP-1 may be involved in the pathological changes of PTC through CD47 in UUO kidneys.
