【摘要】 目的:观察经超声介导的核转移因子κB圈套寡核苷酸作用后大鼠移植肝的肝功能变化、急性排斥的病理情况及受体的生存时间,探讨超声介导的核因子κB圈套寡核苷酸抗大鼠肝移植急性排斥的作用。方法:实验于2005-04/11在南京医科大学第一附属医院动物实验中心完成。①DA大鼠和Lewis大鼠各36只随机分为3组;每组DA大鼠(供体)、Lewis大鼠(受体)各12只,行DA大鼠至Lewis大鼠的原位肝移植。①对照组:对供肝无干预处理。②生理盐水组:生理盐水1mL+核因子κB圈套寡核苷酸100μg灌注供肝,并在冰水浴中经超声(频率2MHz)处理1min。③10%声诺维组:10%声诺维生理盐水1mL+核因子κB圈套寡核苷酸100μg灌注供肝,并在冰水浴中经超声(频率2MHz)处理1min。④检测各组受体大鼠(n=4)术后第1,4,7天的肝功能(天冬氨酸氨基转移酶、丙氨酸转氨酶)变化、病理组织学检查术后第4天移植肝的排斥情况(n=4,按Banff标准进行程度分级,分为0,Ⅰ,Ⅱ,Ⅲ,0级:无排斥证据;Ⅲ级:严重排斥)、统计各组受体大鼠生存率。结果:生理盐水组和10%声诺维组各有1只受体大鼠在术后3d内死亡,未纳入统计。进入结果分析34只。①肝移植术后各组受体大鼠天冬氨酸氨基转移酶、丙氨酸转氨酶术后第1天便有不同程度的升高;10%声诺维组各时间点的天冬氨酸氨基转移酶、丙氨酸转氨酶值明显低于对照组和生理盐水组(P<0.01);对照组和生理盐水组的转氨酶变化差异无显著性。②术后第4天病理组织学检查各组急性排斥情况:10%声诺维组急性排斥较对照组轻(P<0.05),与生理盐水组比较差异无显著性(P>0.05)。③10%声诺维组肝移植术后受体生存时间较对照组和生理盐水组明显延长(P<0.01);对照组与生理盐水组之间术后生存时间差异无显著性。结论:超声和微泡超声造影剂介导的核因子κB圈套寡核苷酸处理大鼠移植肝,能够改善术后移植肝功能、减轻术后移植肝的急性排斥反应、延长受体大鼠的术后生存期。更多还原

【Abstract】 AIM: To observe the influence of transfection of nuclear factor κB decoy oligodeoxynucleotides into donor liver using ultrasound mediator on liver function, pathological changes of acute cellular rejection of liver grafts and survival of recipient rats, and study if transfection of nuclear factor κB decoy oligodeoxynucleotides using ultrasound mediator into donor liver has effect on preventing acute cellular rejection of liver transplantation of rats. METHODS: The experiment was performed in the Animal Central Laboratory, First Affiliated Hospital of Nanjing Medical University from April to November 2005. ①Thirty-six DA rats and 36 Lewis rats were divided into three groups randomly, 12 DA rats (donator) and 12 Lewis rats (recipient) in each group. Orthotopic liver transplantation was adopted in DA rats and Lewis rats: ①Control group: there was no intervention on donor liver. ②Saline group: 100 μg nuclear factor κB decoy oligodeoxynucleotides plus 1 mL saline was infused into the donor liver, followed by ultrasound (2 MHz) exposure for 1 minute in ice water bath. ③10% SonoVue group: 100 μg nuclear factor κB decoy oligodeoxynucleotides plus 1 mL saline containing 10% SonoVue was infused into the donor liver, followed by ultrasound (2 MHz) exposure for 1 minute in ice water bath. ④Blood samples were taken from each group (n=4) for the detection of liver function (aspartate aminotransferase, alanine aminotransferase) at days 1, 4 and 7 after liver transplantation. The samples of liver tissues of each group (n=4, grading according to Banff standard, 0, Ⅰ, Ⅱ, Ⅲ. 0 grade represented no rejection; Ⅲ grade represented severe rejection) were taken to make sections for pathohistological detection at day 4 after liver transplantation. Survival rate of recipient rats was determined. RESULTS: One recipient rat in saline group and one recipient rat in 10% SonoVue group died within 3 days after operation, so thirty-four rats were involved in the result analysis. ①Levels of aspartate aminotransferase and alanine transaminase of recipient rats after liver transplantation increased to different degree at day 1 after operation. Levels of aspartate aminotransferase and alanine transaminase were significantly lower at each time point in the 10% SonoVue group than those in the control group and saline group (P < 0.01). There was no significant difference in aminotransferase between the control group and saline group. ②At day 4 after liver transplantation, the degree of acute cellular rejection with histopathologic examination: The acute cellular rejection in the 10% SonoVue group was light as compared with the control group (P < 0.05). There was no significant difference as compared with the saline group (P > 0.05). ③Survival duration of recipients in the 10% SonoVue group was markedly longer than that in the control group and saline group after liver transplantation (P < 0.01). There was insignificant difference in post-operative live time between the control group and the saline group. CONCLUSION: Transfection of nuclear factor κB decoy oligodeoxynucleotides into the donor liver using ultrasound-SonoVue mediator can ameliorate graft liver function, prevent liver acute cellular rejection and prolong survival duration of recipient rats.更多还原