中国医学前沿杂志（电子版）
中國醫學前沿雜誌（電子版）
중국의학전연잡지（전자판）
CHINESE JOURNAL OF THE FRONTIERS OF MEDICAL SCIENCE(ELECTRONIC VERSION)
2014年
9期
9-14
,共6页
艾娜%谢席胜%夏梦迪%龙琼先%王彦江
艾娜%謝席勝%夏夢迪%龍瓊先%王彥江
애나%사석성%하몽적%룡경선%왕언강
肾缺血再灌注损伤%氧化应激%血小板反应蛋白-1%CD47%免疫荧光双染
腎缺血再灌註損傷%氧化應激%血小闆反應蛋白-1%CD47%免疫熒光雙染
신결혈재관주손상%양화응격%혈소판반응단백-1%CD47%면역형광쌍염
Ischemia-reperfusion injury%Thrombospondin-1%CD47%Reactive oxygen species%Immunolfu-orescence double staining
目的：构建肾缺血再灌注损伤（IRI）大鼠模型，观察血小板反应蛋白-1（TSP-1）及其受体CD47在肾脏的分布及变化，并探讨其在肾IRI发病机制中的作用。方法成年雄性SD大鼠42只，随机分为正常对照组（n＝5）、假手术组（n＝5）和模型组（n＝32），模型组下设IRI后1小时、6小时、12小时、24小时4个时间点（4个小组），每个时间点各8只大鼠。通过阻断肾血流45分钟后再灌注构建肾IRI大鼠模型。观察肾IRI后各组大鼠尿量变化，采用全自动生化分析仪检测各组大鼠的肾功能，通过HE染色了解肾脏病理损害，采用酶联免疫吸附法测定肾组织中羟自由基和丙二醛的浓度。通过肾组织免疫组化及免疫荧光双染，观察TSP-1和CD47蛋白在肾脏中的表达。结果与正常对照组和假手术组相比，模型组大鼠尿量明显减少，血肌酐和尿素氮水平明显升高，差异具有显著性；随着缺血再灌注后时间的延长，肾小管出现不同程度的扩张、变性与坏死。肾间质出现炎性细胞浸润、充血水肿等变化。模型组大鼠肾小管HE染色病理损伤评分较正常组升高，差异具有显著性；IRI肾组织中的羟自由基和丙二醛水平呈逐渐升高趋势。在肾IRI早期，肾小管TSP-1、CD47表达水平即明显增加，至肾IRI 24小时时TSP-1、CD47在残存肾小管中仍有较高表达。通过免疫荧光双染发现，在肾IRI肾小管TSP-1和CD47存在共表达。相关性分析发现羟自由基和丙二醛的浓度与TSP-1及CD47积分光密度呈正相关。结论肾IRI时大鼠肾脏TSP-1和CD47表达水平明显升高，与氧化应激反应密切相关。TSP-1及其受体CD47共同参与了肾IRI过程。
目的：構建腎缺血再灌註損傷（IRI）大鼠模型，觀察血小闆反應蛋白-1（TSP-1）及其受體CD47在腎髒的分佈及變化，併探討其在腎IRI髮病機製中的作用。方法成年雄性SD大鼠42隻，隨機分為正常對照組（n＝5）、假手術組（n＝5）和模型組（n＝32），模型組下設IRI後1小時、6小時、12小時、24小時4箇時間點（4箇小組），每箇時間點各8隻大鼠。通過阻斷腎血流45分鐘後再灌註構建腎IRI大鼠模型。觀察腎IRI後各組大鼠尿量變化，採用全自動生化分析儀檢測各組大鼠的腎功能，通過HE染色瞭解腎髒病理損害，採用酶聯免疫吸附法測定腎組織中羥自由基和丙二醛的濃度。通過腎組織免疫組化及免疫熒光雙染，觀察TSP-1和CD47蛋白在腎髒中的錶達。結果與正常對照組和假手術組相比，模型組大鼠尿量明顯減少，血肌酐和尿素氮水平明顯升高，差異具有顯著性；隨著缺血再灌註後時間的延長，腎小管齣現不同程度的擴張、變性與壞死。腎間質齣現炎性細胞浸潤、充血水腫等變化。模型組大鼠腎小管HE染色病理損傷評分較正常組升高，差異具有顯著性；IRI腎組織中的羥自由基和丙二醛水平呈逐漸升高趨勢。在腎IRI早期，腎小管TSP-1、CD47錶達水平即明顯增加，至腎IRI 24小時時TSP-1、CD47在殘存腎小管中仍有較高錶達。通過免疫熒光雙染髮現，在腎IRI腎小管TSP-1和CD47存在共錶達。相關性分析髮現羥自由基和丙二醛的濃度與TSP-1及CD47積分光密度呈正相關。結論腎IRI時大鼠腎髒TSP-1和CD47錶達水平明顯升高，與氧化應激反應密切相關。TSP-1及其受體CD47共同參與瞭腎IRI過程。
목적：구건신결혈재관주손상（IRI）대서모형，관찰혈소판반응단백-1（TSP-1）급기수체CD47재신장적분포급변화，병탐토기재신IRI발병궤제중적작용。방법성년웅성SD대서42지，수궤분위정상대조조（n＝5）、가수술조（n＝5）화모형조（n＝32），모형조하설IRI후1소시、6소시、12소시、24소시4개시간점（4개소조），매개시간점각8지대서。통과조단신혈류45분종후재관주구건신IRI대서모형。관찰신IRI후각조대서뇨량변화，채용전자동생화분석의검측각조대서적신공능，통과HE염색료해신장병리손해，채용매련면역흡부법측정신조직중간자유기화병이철적농도。통과신조직면역조화급면역형광쌍염，관찰TSP-1화CD47단백재신장중적표체。결과여정상대조조화가수술조상비，모형조대서뇨량명현감소，혈기항화뇨소담수평명현승고，차이구유현저성；수착결혈재관주후시간적연장，신소관출현불동정도적확장、변성여배사。신간질출현염성세포침윤、충혈수종등변화。모형조대서신소관HE염색병리손상평분교정상조승고，차이구유현저성；IRI신조직중적간자유기화병이철수평정축점승고추세。재신IRI조기，신소관TSP-1、CD47표체수평즉명현증가，지신IRI 24소시시TSP-1、CD47재잔존신소관중잉유교고표체。통과면역형광쌍염발현，재신IRI신소관TSP-1화CD47존재공표체。상관성분석발현간자유기화병이철적농도여TSP-1급CD47적분광밀도정정상관。결론신IRI시대서신장TSP-1화CD47표체수평명현승고，여양화응격반응밀절상관。TSP-1급기수체CD47공동삼여료신IRI과정。
Objective To evaluate the effects of TSP-1 and its ligand CD47 in renal ischemia-reperfusion injury (IRI) in the rat model. Method A total of 42 male Sprague-Dawley rats were randomly divided into normal group (n = 5), sham group (n = 5) and IRI group (n = 32). IRI group patients were divided into four time points:1 hour, 6 hours, 12 hours, 24 hours, each time point contain 8 animals. Through blocking bilateral renal blood lfow for 45 minutes in rats to established IRI model. Observed the urine volume of each group. Detected renal function, hematoxylin-eosin staining (HE) and used enzyme-linked immunosorbent method determination the concentration of hydroxyl free radicals and malondialdehyde in kidney tissues. The expression of TSP-1 and CD47 was measured by immunohistochemical and immunolfuorescence double dye. Result Compared with normal group and sham group, urine output were remarkably decreased, and the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were gradually increased in IRI groups. Animals subjected to IRI displayed extensive features of acute tubular damage, including cellular swelling, brush border loss, tubular dilatation, tubule cast formation, and inflammatory cell infiltration in the inner cortex and outer medullary regions. With the extension of the time after reperfusion injury, the concentration of hydroxyl free radicals and malondialdehyde in the kidney tissues were increased. At the early of the ischemia-reperfusion injury, the expression of TSP-1 and CD47 in kidney tissues were gradually increased, at IRI 24 hours the expression were still high in residual renal. Immunolfuorescence double dye found that TSP-1 and CD47 were coexpression in renal IRI renal tubules. Correlation analysis found that the positive rate of hydroxyl free radicals and malondialdehyde with serum creatinine, blood urea nitrogen and renal tubular damage score, the expression of TSP-1 and CD47. Conclusion TSP-1 and CD47 expression signiifcantly increased, it is closely related to the oxidative stress reaction. TSP-1 and CD47 is the early warning indicator of renal tubular injury, participation the early damage of IRI and the progress of kidney disease.
