白细胞介素1受体拮抗剂联合骨髓间充质干细胞移植治疗猪急性肝衰竭

Combined therapy of mesenchymal stem cell transplantation and interleukin 1 receptor antagonist for swine acute liver failure

目的观察白细胞介素1受体拮抗剂（IL-1Ra）联合同种异体骨髓间充质干细胞（MSCs）移植对猪急性肝功能衰竭的协同治疗作用。方法采用D-氨基半乳糖（D-Gal）诱导建立猪急性肝功能衰竭模型。21只中华实验小型猪随机分为3组（n=7），对照组（A）：诱导24h后，门静脉移植生理盐水40ml；单纯MSCs移植组（B）：诱导24h后，门静脉移植8×10^7个MSCs；联合治疗组（C）：诱导24h后，门静脉移植8×10^7个MSCs。分别在移植前6h、移植后1、3d猪耳背大静脉推注IL-1Ra共10mg。移植后4周内定期检测肝功能、病理改变和炎性因子变化并监测移植细胞体内存活转化情况。结果联合治疗组在促进肝功能恢复、改善炎症环境和减轻肝脏损伤方面较对照组和单纯MSCs移植组差异有统计学意义（P〈0．05）；免疫组织化学显示联合治疗组肝细胞增殖率在1周后分别为67．70±9．47、71．80±9．45和40．04±4．60，显著高于对照组和单纯移植组，与对照组差异有统计学意义（P〈0．05）；免疫组织化学和荧光染色法均可见联合治疗组干细胞植入数要高于单纯移植组。IL-1Ra在改善肝衰竭炎症内环境中发挥重要作用，通过对宿主炎症反应的有效控制可以显著提高移植细胞的植入率，促进肝细胞再生。结论IL-1Ra联合MSCs移植治疗具有协同作用，可以显著改善D-Gal诱导的猪急性肝衰竭的肝功能指标，促进肝组织再生。

Objective To evaluate synergic effects of allogeneic mesenchymal stem cells （MSCs） transplantation and interleukin 1 receptor antagonist （IL-1Ra） on swine acute liver failure. Methods Chinese experimental mini swine were given D-galactosamine to establish models of acute liver failure. Twentyone pigs were randomly divided into three groups （n =7 each）. In the control group （A）, the normal saline was injected into the liver via the portal vein after induction for 24 h; In the MSCs transplantation group （B）, 8 × 10^7 MSCs （in 40 ml PBS） were injected into the liver via the portal vein after induction for 24 h; In combined therapy group （C）, 8 × 10^7 MSCs were injected into the liver via the portal vein after induction for 24 h, followed by injection of rhIL-1Ra （10 mg/pig） via the ear vein at 6 h before, and one day and three days after transplantation respectively. Liver function, serum inflarmnation and pathological changes were measured. The fate of MSCs was also observed. Results The biochemical assay, the serum inflammation level and pathological changes in group C were all greatly different from those in group A and group B （P 〈 0. 05 ）. The proliferation rate of hepatocytes in group C was 67.70 ± 9, 47, 71.80 ± 9. 45, and 40. 04 ± 4. 60 one week after transplantation, significantly higher than in group A and group B （P 〈0. 05）. The number of MSCs in group C was greater than in group B. Hepatocytes regeneration were associated with the kine- Sis changes of inflammation. Our findings suggest IL-1Ra as an important player in the repair of acute liver failure （ALF）. Improving homeostasis of liver failure contributed to increased hepatic engraftment and hepatocytes proliferation. Conclusion The MSCs transplantation is useful for ALF partially. The combined therapy shows cooperative effects.