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大鼠小体积肝脏移植后早期肝内细胞因子的变化

Changes of early intragraft cytokines following small-for-size liver transplantation in rats

摘要:

目的 研究不同冷缺血条件下大鼠小体积肝移植(30%标准体积)后早期肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)的变化,及其与肝脏再生的关系.方法 建立Lewis大鼠30%标准体积的原位肝移植模型.根据供肝在UW液中冷保存时间的不同,将受者分为3组:冷缺血1 h组、冷缺血8 h组和冷缺血16 h组,每组均为20只.观察受者存活情况至术后第7天,并分别在移植肝恢复血流后90 min、1 h、2 h、4 h和7 d收集样本,检测移植肝组织中TNF-α和IL-6表达情况,肝细胞DNA的合成情况,进行移植肝的形态学观察.结果 大鼠肝移植手术成功率均为100%.移植后第7天,冷缺血1 h和8 h组受鼠的存活率均为100%.冷缺血16 h组受鼠的存活率较低,移植后第7天无受鼠存活.冷缺血1 h组TNF-α和IL-6的表达水平较低,冷缺血8 h组和冷缺血16 h组TNF-α和IL-6的表达则高于冷缺血1 h组(F=58.81和F=184.12,P<0.05).冷缺血8 h组和冷缺血16 h组间TNF-α和IL-6的表达的差异无统计学意义.冷缺血1 h组增殖细胞数目明显高于冷缺血8 h组,差异有统计学意义(t=5.59,P<0.05).移植术后24h,冷缺血1 h组移植肝有轻度的组织学损伤;冷缺血8 h组移植肝有轻度的窦状隙扩张和轻度的炎症;冷缺血16 h组移植肝有局部淤血,存在肝细胞崩解和坏死等改变.结论 在小体积肝移植后早期,TNF-α和IL-6的上调表达对肝脏再生有重要意义.不同冷缺血时间的小体积肝脏移植物内存在早期启动肝脏再生的信号.

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Objective To explore the effect of different cold ischemia (CI) time on the changes of early intragraft cytokines (TNF-α and IL-6) and their correlation with regeneration after small-for-size (30% standard volume) liver transplantation (LT) in rats. Methods A model of Lewis rat 30% standard volume liver transplantation was established. The rats were divided into 3 groups: 1 h CI group, 8 h CI group and 16 h CI group (n= 20 in each group) according to the CI time of donor livers stored in UW solution. Survival rate at day 7 after LT in each group was recorded and specimens were collected at predetermined intervals from 90 min, 1, 2, 4 h and 7 day post-reperfusion. Expression patterns of TNF-αand IL-6 were determined and compared in 30% liver grafts with different CI time following transplantation. Progression of DNA synthesis of hepatocytes was confirmed by BrdU uptake. Morphological assessment of the liver grafts was also made. Results Operative success rate of LT was 100% in all groups. Mean survival rate in 1 h and 8 h CI group was 100% respectively (> 7 day). Mean survival rate in 16 h CI group was low, and no recipient animals survived on the postoperative day 7. Compared with 1 h CI group, IL-6 expression in 30% liver grafts stored for 8 h and 16 h was markedly increased post-transplantation (F=184.12, P<0.05). TNF-α expression in 30% liver grafts in 8 h and 16 h CI groups was markedly increased post-reperfusion (F=58.81, P< 0.05). There was no statistically significant difference in the expression of TNF-α and IL-6 between 8 h CI and 16 h CI groups. Number of positively stained nuclei in 1 h CI group was greater than that in 8 h CI group at 24 h after transplantation (t=5.59, P<0.05). At 24 h after transplantation histology showed mild injury in grafts of CI 1 h group. CI 8 h led to mild sinusoidal dilatation and inflammation.Grafts stored for 16 h demonstrated focal congestion, hepatocyte collapse and necrosis. Conclusion At the early stage of small-for-size liver transplantation, up-regulation of cytokines (TNF-α and IL-6) is an important factor in the liver regeneration. The early initiating signals for the regenerative response are present in the small-for-size liver grafts with different CI time.

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