依达拉奉对不同冷缺血时间大鼠移植供肝JAK2/STAT3信号通路的影响

摘要

Objective To investigate the effect of edaravone on the JAK2/STAT3 signaling pathway after ischemia-reperfusion injury in donor rat liver under different cold ischemia times. Methods A total of 102 SD rats were randomly divided into sham operation group,control group and experimental group. Six rats were in sham operation group with free liver operation and no transplantation. Forty-eight rats were in control group and experimental group respectively, and divided into subgroups according to the different cold ischemia times (30 min, 6 h, 12 h and 18 h). There were 6 donors and 6 recipients in each group. The rat model of orthotopic liver transplantation was established by modified"two cuff method". All the donors were perfused by abdominal aorta and the warm ischemia time was 3-5 min. After different cold ischemia times, the experimental group was treated with edaravone (3 mg/kg) at 5 min before the opening of the new hepatic artery, and control group was injected with 3 mg/kg saline. Recipients of each group were sacrificed after 6 h. Finally, real-time fluorescence quantitative PCR was used to analyze the relative expression of JAK2/STAT3 mRNA of donor liver. Results The GAPDH gene and JAK2/STAT3 were well amplified. Under the same cold ischemia time, compared with the control group, the relative expression of JAK2/STAT3 was significantly decreased in the experimental group (P<0.05). With the prolongation of cold ischemia time, the relative expressions of JAK2 and STAT3 mRNA showed a decreasing trend in control group and experimental group, while the relative expression of JAK2 mRNA increased first and then decreased in the experimental group (P<0.05). Conclusion Edaravone has a protective effect on transplanted donor liver during different cold ischemia times, and extends the cold ischemia time for 18 h, which may be related to the inhibition of JAK2/STAT3 signal transduction pathway.%目的 探讨依达拉奉对不同冷缺血时间大鼠移植供肝缺血再灌注损伤(IRI)后JAK2/STAT3信号通路的影响.方法 SD大鼠102只,随机分为假手术组、对照组及实验组.假手术组6只,游离肝脏,不进行移植;对照组及实验组各48只,2组各自按照不同冷缺血时间30 min、6 h、12 h、18 h分为4个处理亚组,每处理组供体、受体各6只.采用改良的"二袖套法"建立大鼠原位肝移植模型,供体采用腹主动脉灌注法,热缺血时间为3~5 min.供肝在不同冷缺血时间后,实验组分别于新肝血管开放前5 min经鼠尾静脉注射依达拉奉3 mg/kg;对照组分别注射生理盐水3 mg/kg.各组受体均于6 h后处死取材.采用实时荧光定量PCR(RT-PCR)检测受体供肝组织中JAK2和STAT3 mRNA的相对表达量.结果 内参基因GAPDH与检测基因JAK2、STAT3扩增良好;在同一冷缺血时间,与对照组比较,实验组大鼠供肝基因JAK2/STAT3相对表达量均明显降低(P<0.05);随冷缺血时间延长,对照组JAK2、STAT3 mRNA和实验组STAT3 mRNA相对表达量均呈逐渐降低趋势,而实验组JAK2 mRNA相对表达量呈先增后降趋势(P<0.05).结论 依达拉奉对不同冷缺血时间移植供肝具有一定的保护作用.依达拉奉可延长供肝冷缺血时间至18 h,其机制可能与抑制肝细胞JAK2/STAT3信号转导通路有关.