shRNA沉默CⅡTA基因表达对大鼠肝移植急性排斥反应的影响
首发时间:2008-03-25
摘要:目的:探讨经门静脉注射CⅡTA-shRNA质粒方法干预供体移植物对大鼠原位肝移植急性排斥反应的影响。方法:建立DA大鼠至Lewis大鼠的高应答原位肝移植模型;采用经门静脉注射质粒方法干预供体大鼠肝脏,并设置盐水对照组、阴性质粒对照组和环孢素A治疗组,移植术后第4天随机选取6只受体大鼠取出移植肝脏,病理学检查评定排斥反应分级,荧光实时定量PCR方法检测标本中CⅡTA、MHC-Ⅱ、IL-2和IFN-γ的表达水平变化,免疫组化方法检测MHC-Ⅱ抗原表达变化,其余6只用于观察存活期。结果:与对照组相比,CⅡTA-shRNA质粒干预组大鼠存活时间显著延长(P<0.01),排斥反应病理分级显著降低(P<0.01),CⅡTA、MHC-Ⅱ、IL-2和IFN-γ的mRNA表达水平及MHC-Ⅱ抗原表达水平均显著降低(P<0.01)。结论:采用经门静脉注射CⅡTA-shRNA质粒方法干预供体移植物可明显减轻肝移植急性排斥反应。
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The effect of shRNA interfering CⅡTA on acute rejection in rat liver transplantation
Abstract:Objective: To explore the effect of shRNA interfering CⅡTA in graft by portal vein injection on acute rejection reaction in rat liver transplantation. Methods: The high responder model of rat orthotopic liver transplantation was established. There were 4 groups: normal saline control group, pHK-shRNA control group, pCⅡTA-shRNA treating group and ciclosporin A (CsA) treating group. Graft was pretreated by portal vein injection of plasmid. 6 recipients in each group were randomly chosen and sacrificed 4 days after transplantation and the remaining 6 recipients were monitored for survival. The data of pathological rejection grading, MHC-Ⅱ expression level, transcription levels of CⅡTA, MHC-Ⅱ, IL-2 and IFN-γ in liver specimens were collected by pathological examination, immunohistochemistry staining and real time QRT-PCR. Results: Compared to control groups, the survival time of pCⅡTA-shRNA group extended significantly (P<0.01), pathological rejection grading significantly depressed (P<0.01), MHC-Ⅱ expression level significantly decreased (P<0.01) and transcription levels of CⅡTA, MHC-Ⅱ, IL-2 and IFN-γ significantly decreased (P<0.01). Conclusion: Acute rejection reaction can be significantly relieved and survival time is extended by pretreatment of graft and donor with portal vein injection of CⅡTA-shRNA plasmid.
Keywords: CⅡTA shRNA liver transplantation acute rejection reaction
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shRNA沉默CⅡTA基因表达对大鼠肝移植急性排斥反应的影响
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