摘要
目的在建立大鼠纤维化肝移植模型的基础上,检测肝星状细胞的激活,探讨Rho- ROCK信号通路及血管内皮生长因子(VEGF)的表达在小肝移植后供肝损伤中所起的作用。方法建立大鼠纤维化肝移植模型;分全肝移植组和小肝移植组。观察两组7 d生存率;移植后不同时间点取样本,应用α-SMA抗体进行免疫组织化学染色以测定肝星状细胞(Hepatic Stellate Cell)的激活程度;Western blot检测Rho、ROCK I及VEGF的表达。结果成功建立大鼠纤维化肝移植模型; 全肝移植组和小肝移植组7 d生存率分别为58．3％(7／12)和8．3％(1／12,P<0．05);在同一组内不同时间点上,α-SMA的表达呈上升趋势;在各时间点上,小肝移植组α-SMA的表达均高于同一时间点全肝移植组,表明小肝移植组HSCs激活的程度总体上高于全肝移植组;小肝移植组Rho、ROCK I和VEGF的表达均高于同一时间点的全肝移植组。结论由于Rho—ROCK信号通路导致的肝星状细胞激活在大鼠纤维化肝移植中小体积供肝损伤有显著意义。
Objective To investigate the activation of hepatic stellate cells in a rat liver transplantation model using whole or small-for-size grafts and cirrhotic recipients. The possible underlying mechanism related to Rho-ROCK-VEGF signaling pathway was also studied. Methods A rat orthotopic liver transplantation model using normal liver grafts （100 % and 50 % of the recipient rat liver） was applied in cirrhotic rat recipients. The cirrhotic rat model was induced by subcutaneous injection of CC14 for 8 weeks. Recipients were sacrificed at 6, 24 and 48 h after liver transplantation. The liver tissues were sampled for morphological examination, hepatic stellate cell （HSC） activation （α-SMA expression）, VEGF expression, Rho and ROCK signaling pathways by immunostaining and Western blot. Liver function and 7-day survival rates were compared between the rats implanted with whole and small-for-size liver grafts. Results The 7-day graft survival rate of smaU-for-size grafts was significantly lower than that of whole grafts [8.3% （1/12） vs 58.3% （7/12）, P〈 0.05]. The deterioration of liver function was mainly found in small-for-size grafts, especially at 6 h after reperfusion. The irttragraft expression of α-SMA in the small-for-size graft group was significantly higher than that of the whole graft group during the first 48 h after liver transplantation. The intragraft protein levels of Rho, ROCKⅠ and VEGF were also significantly elevated in the small-for-size graft group compared to those of the whole graft group at different time points after reperfusion. Conclusion Significant activation of hepatic steUate cells through up-regulation of Rho-ROCK-VEGF signaling pathway might play an important role in small-for-size graft injury in a cirrhotic rat liver transplantation model.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第12期1452-1454,i0021,共4页
Chinese Journal of Experimental Surgery
