骨髓间充质干细胞移植对急性肝衰竭大鼠Caspase1和IL-18表达的影响

Effect of bone marrow mesenchymal stem cell transplantation on expression of Caspase1 and IL-18 in rats with acute hepatic liver failure

目的:探讨骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)移植治疗急性肝衰竭(acute liver failure,ALF)大鼠的疗效、移植途径.方法:用D-氨基半乳糖(D-galactosamine,D-GalN)/脂多糖(lipopolysaccharide,LPS)诱导大鼠ALF模型.72只大鼠随机分为ALF对照组、尾静脉移植组、门静脉移植组.于BMSCs移植后24、72、120、168 h取血清和肝组织,分别检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST);采用TUNEL法检测肝细胞凋亡;采用免疫组织化学、Western blot方法检测肝组织中半胱氨酸蛋白酶1(cysteinecontaining aspartate-specific proteases 1,Caspase1)和白介素18(interleukin-18,IL-18)蛋白的表达情况.结果:ALF对照组大鼠血清ALT、AST水平随病程逐渐升高.在移植后120、168 h,BMSCs移植组的血清ALT、AST与对照组相比差别有统计学意义(P<0.05);尾静脉、门静脉移植组细胞凋亡指数分别为28.17%±17.08%、20.67%±12.68%、19.67%±11.82%、13.00%±6.84%,与对照组相比差异有统计学意义(P<0.01).BMSCs移植组Caspase1、IL-18蛋白表达水平逐渐降低,在移植后120、168 h与对照组相比差异有统计学意义(P<0.05);且二者的表达有明显的相关性(P<0.01).尾静脉、门静脉移植途径对ALF大鼠均有治疗作用,但二种途径相比差异无统计学意义(P>0.05).结论:BMSCs能够改善ALF肝衰竭大鼠的肝功能,抑制肝细胞凋亡、降低Caspase1、IL-18蛋白水平.Caspase1和IL-18在肝衰竭的发病过程中起重要作用;Caspase1和IL-18可望成为急性肝衰竭的预测因子和未来的治疗靶点.

AIM： To investigate the effect of bone marrow mesenchymal stem cell （BMSC） transplantation in the treatment of acute liver failure （ALF） inrats and optimize the administration route METHODS： D-galactosamine （D-GalN）/lipo- polysaccharide （LPS） were used to make a rat model of experimental ALF. Seventy-two rats were randomly divided into an ALF group, a tail vein group, and a portal vein transplanta- tion group. At different time points （24, 72, 120, and 168 h） after BMSC transplantation, serum ALT and AST were detected. TUNEL assay was applied to detect hepatocyte apoptosis. Immu- nohistochemistry and Western blot were per- formed to detect the expression of Caspasel and IL-18 proteins in liver tissue. RESULTS： Serum levels of ALT and AST in the ALF group were gradually increased with the progression of the disease. Compared with the ALF group, significant improvement of liver function parameters and histological findings was observed in the transplantation group 120 and 168 h after transplantation （P 〈 0.05 for both）. The apoptosis indexes in the tail vein group and portal vein transplantation group （120 h： 28.17% ± 17.08%, 20.67% ± 12.68%; 168 h： 19.67 % ± 11.82%, 13.00 %± 6.84% ） were significantly lower than those in the ALF group （P 〈 0.01 for all）. The expression levels of Caspasel and IL-18 proteins in the two transplantation groups were decreased significantly at 120 and 168 h com- pared with the ALF group （P 〈 0.05 for both）. Both administration routes had a therapeutic effect against ALF in rats, but no significant dif- ference was observed between them. CONCLUSION： BMSCs can improve the liver function, inhibit hepatic apoptosis and reduce the levels of Caspasel and IL-18 proteins in ALF rats. Caspasel and IL-18 play an important role in the pathogenesis of ALF and are expected to be pre- dictors of ALF and future therapeutic targets.