中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2012年
12期
737-741
,共5页
唐华%万英%张春梅%陈蓉%郭庆喜%曾维诚
唐華%萬英%張春梅%陳蓉%郭慶喜%曾維誠
당화%만영%장춘매%진용%곽경희%증유성
大鼠%肾移植%移植物排斥%载脂蛋白M%NF-κB
大鼠%腎移植%移植物排斥%載脂蛋白M%NF-κB
대서%신이식%이식물배척%재지단백M%NF-κB
Rats%Kidney transplantation%Graft rejection%Apolipoprotein M%NF-κB
目的 观察大鼠肾移植后载脂蛋白M(apoM)的表达,探讨apoM在急性排斥反应(AR)中的作用及机制.方法 以SD大鼠和Wistar大鼠作为供鼠,Wistar大鼠作为受鼠,建立同系和同种大鼠原位肾移植模型.实验共分3组,对照组取同系移植受鼠,AR组和PDTC组取同种移植受鼠,其中PDIC组受鼠术后0.5 h注射核因子-κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸盐(PDTC),其余2组注射等量生理盐水.术后1、3、5和7d,取各组血清和移植肾组织进行检测,采用蛋白质印迹法检测NF-κB P65和apoM蛋白的表达,采用实时聚合酶链反应法检测apoM、穿孔素和颗粒酶BmRNA的表达,并对各检测指标进行相关性分析.结果 术后各时间点,AR组和PDTC组apoM、穿孔素和颗粒酶BmRNA的表达水平均明显高于同期对照组(P＜0.01),而PDTC组3种mRNA的表达水平均较同期AR组显著降低(P＜0.01).AR组和PDTC组apoM和NF-κBP65蛋白的表达均较同期对照组明显增高(P＜0.01),PDTC组2种蛋白的表达水平均较同期AR组显著降低(P＜0.01).经相关性分析,apoM蛋白与NF-κB P65蛋白的表达呈直线正相关(r=0.469、P＜0.05),apoM mRNA与穿孔素和颗粒酶BmRNA的表达均呈正相关(r=0.731、P＜0.0l,r=0.514,P＜0.05).结论 同种肾移植后,受鼠移植肾组织内apoM的表达显著上调,其可能通过NF-κB的介导参与AR的发生.
目的 觀察大鼠腎移植後載脂蛋白M(apoM)的錶達,探討apoM在急性排斥反應(AR)中的作用及機製.方法 以SD大鼠和Wistar大鼠作為供鼠,Wistar大鼠作為受鼠,建立同繫和同種大鼠原位腎移植模型.實驗共分3組,對照組取同繫移植受鼠,AR組和PDTC組取同種移植受鼠,其中PDIC組受鼠術後0.5 h註射覈因子-κB(NF-κB)抑製劑吡咯烷二硫代氨基甲痠鹽(PDTC),其餘2組註射等量生理鹽水.術後1、3、5和7d,取各組血清和移植腎組織進行檢測,採用蛋白質印跡法檢測NF-κB P65和apoM蛋白的錶達,採用實時聚閤酶鏈反應法檢測apoM、穿孔素和顆粒酶BmRNA的錶達,併對各檢測指標進行相關性分析.結果 術後各時間點,AR組和PDTC組apoM、穿孔素和顆粒酶BmRNA的錶達水平均明顯高于同期對照組(P＜0.01),而PDTC組3種mRNA的錶達水平均較同期AR組顯著降低(P＜0.01).AR組和PDTC組apoM和NF-κBP65蛋白的錶達均較同期對照組明顯增高(P＜0.01),PDTC組2種蛋白的錶達水平均較同期AR組顯著降低(P＜0.01).經相關性分析,apoM蛋白與NF-κB P65蛋白的錶達呈直線正相關(r=0.469、P＜0.05),apoM mRNA與穿孔素和顆粒酶BmRNA的錶達均呈正相關(r=0.731、P＜0.0l,r=0.514,P＜0.05).結論 同種腎移植後,受鼠移植腎組織內apoM的錶達顯著上調,其可能通過NF-κB的介導參與AR的髮生.
목적 관찰대서신이식후재지단백M(apoM)적표체,탐토apoM재급성배척반응(AR)중적작용급궤제.방법 이SD대서화Wistar대서작위공서,Wistar대서작위수서,건립동계화동충대서원위신이식모형.실험공분3조,대조조취동계이식수서,AR조화PDTC조취동충이식수서,기중PDIC조수서술후0.5 h주사핵인자-κB(NF-κB)억제제필각완이류대안기갑산염(PDTC),기여2조주사등량생리염수.술후1、3、5화7d,취각조혈청화이식신조직진행검측,채용단백질인적법검측NF-κB P65화apoM단백적표체,채용실시취합매련반응법검측apoM、천공소화과립매BmRNA적표체,병대각검측지표진행상관성분석.결과 술후각시간점,AR조화PDTC조apoM、천공소화과립매BmRNA적표체수평균명현고우동기대조조(P＜0.01),이PDTC조3충mRNA적표체수평균교동기AR조현저강저(P＜0.01).AR조화PDTC조apoM화NF-κBP65단백적표체균교동기대조조명현증고(P＜0.01),PDTC조2충단백적표체수평균교동기AR조현저강저(P＜0.01).경상관성분석,apoM단백여NF-κB P65단백적표체정직선정상관(r=0.469、P＜0.05),apoM mRNA여천공소화과립매BmRNA적표체균정정상관(r=0.731、P＜0.0l,r=0.514,P＜0.05).결론 동충신이식후,수서이식신조직내apoM적표체현저상조,기가능통과NF-κB적개도삼여AR적발생.
Objective To investigate the expression of apolipoprotein M in rats after renal transplantation and its role and action mechanism in acute rejection (AR).Methods The kidney transplantation model in rats were established.Male SD and Wistar rats were used as donors,and Wistar rats as recipients.Three groups were designated:control group (syngeneic transplantation,n =24,recipients were treated with normal saline i.p.); AR group (allogeneic transplantation,n =24,recipients were treated with normal saline i.p.); PDTC group [allogeneic transplantation,n =24,recipients were treated with NF-κB inhibitor-pyrrolidine dithiocarbamat (PDTC) i.p.].The renal grafts were drawn at day 1,3,5 and 7 post-transplantation,and the expression levels of NF-κB P65 and apoM protein were detected by using Western blotting,and those of apoM,perforin and granzyme B mRNA were by using real-time PCR.The correlation of apoM and NF-κB,apoM and perforin,and apoM and granzyme B was respectively analyzed.Results As compared with control group,the expression levels of apoM,perforin and granzyme B mRNA in AR and PDTC groups were dramatically up-regulated at each time point (P＜0.01),and those in PDTC group were significantly lower than those in AR group (P＜0.01).The expression levels of apoM and NF-κB protein in AR group were both distinctly higher than those in control group (P＜0.01),and those in PDTC group were markedly lower than those in AR group (P＜0.01).A significantly positive correlation was found between the expression of apoM and NF-κB protein (r =0.469,P＜0.05).And the expression of apoM mRNA was positively correlated to perforin and granzyme B mRNA (r =0.731,P＜0.01 ; r =0.514,P＜0.05).Conclusion The expression of apoM is obviously up-regulated in renal grafts of rats,which may take part in the pathogenesis of AR via NF-κB.
