中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2008年
7期
516-519
,共4页
武聚山%夏仁品%卢实春%张毅%娄金丽%李宁
武聚山%夏仁品%盧實春%張毅%婁金麗%李寧
무취산%하인품%로실춘%장의%루금려%리저
肝移植%移植物排斥%OX40%小干扰RNA
肝移植%移植物排斥%OX40%小榦擾RNA
간이식%이식물배척%OX40%소간우RNA
Liver transplantation%Graft rejection%OX40%siRNA
目的 探讨RNA干扰阻断OX40-OX40L共刺激通路对大鼠移植肝存活时间的影响.方法 制作原位肝脏移植物大鼠模型,供体DA大鼠,受体Lewis大鼠.移植术毕时将5 ml含5×109pfu的pLVTHM-OX40-siRNA重组慢病毒经阴茎背静脉在10秒内注射受鼠体内,术后观察受鼠存活时间,移植肝脏进行组织病理学检查,采用ELISA法检测大鼠外周血IL-2、IFN-γ的水平,并进行受者大鼠脾细胞对供者大鼠脾细胞的混和淋巴细胞反应(MLR). 结果转染组大鼠肝脏移植物的存活时间为(74.0±9.3)d,明显长于对照组(7.3±0.5)d和未转染组(7.5±0.5)d;转染组移植肝组织炎细胞浸润、间质水肿、肝组织坏死程度较对照组和未转染组减轻;耐受组大鼠脾脏中T淋巴细胞刺激T淋巴细胞增殖的能力较对照组降低(t=25.1,P＜0.01);移植术后第7 d,转染组血清IL-2浓度为(46±8.4)pg/ml,IFN-γ浓度为(202.7±14.6)pg/ml,均显著低于对照组和未转染组(分别t=176.4,45.5,P＜0.01).结论 转染靶向OX40的siRNA,可通过阻断OX40-OX40L共刺激通路,抑制大鼠肝脏移植后的排斥反应,延长大鼠移植肝的存活时间.
目的 探討RNA榦擾阻斷OX40-OX40L共刺激通路對大鼠移植肝存活時間的影響.方法 製作原位肝髒移植物大鼠模型,供體DA大鼠,受體Lewis大鼠.移植術畢時將5 ml含5×109pfu的pLVTHM-OX40-siRNA重組慢病毒經陰莖揹靜脈在10秒內註射受鼠體內,術後觀察受鼠存活時間,移植肝髒進行組織病理學檢查,採用ELISA法檢測大鼠外週血IL-2、IFN-γ的水平,併進行受者大鼠脾細胞對供者大鼠脾細胞的混和淋巴細胞反應(MLR). 結果轉染組大鼠肝髒移植物的存活時間為(74.0±9.3)d,明顯長于對照組(7.3±0.5)d和未轉染組(7.5±0.5)d;轉染組移植肝組織炎細胞浸潤、間質水腫、肝組織壞死程度較對照組和未轉染組減輕;耐受組大鼠脾髒中T淋巴細胞刺激T淋巴細胞增殖的能力較對照組降低(t=25.1,P＜0.01);移植術後第7 d,轉染組血清IL-2濃度為(46±8.4)pg/ml,IFN-γ濃度為(202.7±14.6)pg/ml,均顯著低于對照組和未轉染組(分彆t=176.4,45.5,P＜0.01).結論 轉染靶嚮OX40的siRNA,可通過阻斷OX40-OX40L共刺激通路,抑製大鼠肝髒移植後的排斥反應,延長大鼠移植肝的存活時間.
목적 탐토RNA간우조단OX40-OX40L공자격통로대대서이식간존활시간적영향.방법 제작원위간장이식물대서모형,공체DA대서,수체Lewis대서.이식술필시장5 ml함5×109pfu적pLVTHM-OX40-siRNA중조만병독경음경배정맥재10초내주사수서체내,술후관찰수서존활시간,이식간장진행조직병이학검사,채용ELISA법검측대서외주혈IL-2、IFN-γ적수평,병진행수자대서비세포대공자대서비세포적혼화림파세포반응(MLR). 결과전염조대서간장이식물적존활시간위(74.0±9.3)d,명현장우대조조(7.3±0.5)d화미전염조(7.5±0.5)d;전염조이식간조직염세포침윤、간질수종、간조직배사정도교대조조화미전염조감경;내수조대서비장중T림파세포자격T림파세포증식적능력교대조조강저(t=25.1,P＜0.01);이식술후제7 d,전염조혈청IL-2농도위(46±8.4)pg/ml,IFN-γ농도위(202.7±14.6)pg/ml,균현저저우대조조화미전염조(분별t=176.4,45.5,P＜0.01).결론 전염파향OX40적siRNA,가통과조단OX40-OX40L공자격통로,억제대서간장이식후적배척반응,연장대서이식간적존활시간.
Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat.Methods siRNA-expression vectors were constructed to targeting OX40.3～5 minutes before DA to Lewis orthotopic liver transplantation was performed,5×109 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline(PBS)and inlected intravenously into recipient Lewis rat over a period of 10 seconds.Serum IL-2 and IFN-γ levels were assayed by ELISA,and mix lymphocyte response(MLR)were tested by 3H-thymidine.Results The survival time of recipients in siRNA treatment group(74.0±9.3)was significantly longer than that in control group[(7.3±0.5)days].In experiment group,the inflammatory cell infihration and liver tissue structure destruction were very slight.The concentration of serum IL-2 was much lower in siRNA treatment group[(46±8.4)pg/ml]than that in control group[(286.5±14.6)pg/ml].Meanwhile,the concentration of serum IFN-γ was much lower in siRNA treatment group [(202.7±14.6)pg/ml]than that in control group[(1682.7±87.9)pg/ml].Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway.hence inhibit the rejection of liver allograft.
