ALPPS一期术后刺激残肝再生及其相关因素的研究

Liver regeneration and related factors after first step of ALPPS surgery in SD rats

目的通过建立联合肝脏分隔和门静脉结扎二步肝切除术模型(associated liver partition and portal vein ligation for staged hepectomy,ALPPS)来探讨骨髓来源肝血窦内皮祖细胞(bone marrow liver sinusoidal endothelial cell progenitor cells,BM SPCs)在ALPPS一期术后肝再生中的作用。方法选取180~200 g健康SD雄性成年大鼠72只,按照随机数字表法分为ALPPS组、门静脉结扎组(portal vein ligation,PVL)和假手术组,分别在术后1、2、4、7 d检测各组大鼠保留侧肝脏体质量比,血清HGF表达水平,肝脏组织BM SPCs表面标记物CD133、CD45、CD31,以及肝功能和肝组织病理学,并对结果进行统计分析。结果 (1)相比于PVL组,ALPPS组在术后Ki-67阳性细胞更多,保留侧肝脏体质量比更高(P<0.05);(2)ALPPS组血清中谷丙转氨酶、谷草转氨酶和总胆汁酸在术后第1天显著升高(P<0.05),第4天与PVL组比较,差异无统计学意义(P>0.05);(3)ALPPS组血清中的HGF在术后第1天达到峰值,明显高于PVL组,从第2天起开始下降,第4天开始有所回升(P<0.05);(4)ALPPS组结扎侧的BM SPCs表面标志物在术后1、2、4、7 d呈阳性,而ALPPS组的保留侧及其PVL组、Sham组均呈阴性。结论 BM SPCs参与了ALPPS术后肝脏的快速再生。

Objective To establish a stable animal model of associated liver partition and portal vein ligation for staged hepatectomy （ALPPS）, and to explore whether bone marrow liver sinusoidal endothelial cell progenitor cells （ BM SPCs ） participate in the regeneration of residual liver after the first step of ALPPS surgery. Methods A total of 72 SD rats （male, adult, weighing 180 -200 g） were randomly divided to ALPPS group （ n = 24）, portal vein ligation （PVL） group （ n = 24） and sham operation group （ n = 24 ）. At 1, 2, 4 and 7 d after the operation, the ratio of residual liver weight to body weight, serum level of hepatocyte growth factor （HGF） , liver expression of BM SPCs surface markers CD133, CD45 and CD31, and liver function and pathological changes were observed and detected, and the results were compared among the groups. Results （1）Compared with the PVL group, the ALPPS group had more Ki-67 positive cells and higher ratio of residual liver weight to body weight （P〈0.05）. （2） The serum levels of alanine aminotransferase （ALT）, glutamic-oxalacetic transaminease （AST） and total bile acid （TBIL） in the ALPPS group were increased significantly （ P 〈 0.05 ） from the first day after surgery, and they had no differences with those of the PVL group at the 4th day （ P 〉 0.05 ）. （3） The serum HGF level in the ALPPS group reached a peak in the first day after surgery, significantly higher than that of the PVL group （ P 〈 0.05 ） , and then reduced at the second day but slightly elevated from the 4th day （ P 〈 0.05 ）. （4）Immunofluorescence assay showed that CD133, CD45 and CD31 were positive in the ligated side at 1, 2, 4 and 7 d after ALPPS operation, whereas were negative in the residual side, PVL group and Sham group. Conclusion BM SPCs take part in the rapid regeneration after ALPPS.