Notch-Hif-1α信号通路参与调控大鼠肝再生过程的研究  

Study on the Notch-Hif-1α Signaling Pathway in Liver Regeneration

作　　者：李博伟[1,2] 李霄[1] 张卓超[1] 孙伟[1] 马奔[1] 牛钢[2] 岳树强[1] 

机构地区：[1]第四军医大学西京医院肝胆外科,陕西西安710032 [2]陕西省宝鸡市中医医院外二科,陕西宝鸡721000

出　　处：《现代生物医学进展》2016年第15期2806-2810,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81370556)

摘　　要：目的:探讨使用γ分泌酶抑制剂Fli-06特异性阻断Notch信号通路后,大鼠肝部分切除术后肝再生的情况并初步阐明Notch-Hif-1α信号通路调控肝再生的可能机制。方法:取SD大鼠分为对照(生理盐水注射组,n=24)和抑制剂组(γ分泌酶抑制剂注射组,n=24)。给予药物处理后,两组分别施行大鼠肝部分切除术,术后0 d,1 d,3 d,5 d,每个时间点分别留取对照组(n=6)及抑制剂组(n=6)再生的肝组织,并检测相应肝重体重比,免疫组化法检测再生肝PCNA(增殖细胞核抗原,Proliferation Cell Nuclear Antigen)表达,RT-PCR检测再生肝组织中的Notch1、Hes1、VEGF m RNA的变化,Western-Blot法检测NICD(Notch胞内段,Notch Intracellular Domain)、Hif-1α(低氧诱导因子-1α,Hypoxia Inducible Factor-1α)蛋白在肝再生过程的变化情况。结果:1、肝部分切除术后3 d和5 d,抑制剂组肝重体重比明显低于对照组差异有统计学意义(P<0.05);2、免疫组织化学染色结果提示:抑制剂组再生肝PCNA阳性细胞率在术后1 d,3 d,5 d均明显低于对照组(P<0.05);3、Western blot结果表明:NICD和Hif-1α蛋白水平明显低于对照组(P<0.05);同时RT-PCR结果提示:抑制剂组Hes1的m RNA表达量术后1 d,3 d明显低于对照组,差异具有统计学意义(P<0.05)。同时,抑制剂组VEGF m RNA水平在术后3 d,5 d明显低于对照组,差异具有统计学意义(P<0.05)。结论:在大鼠肝部分切除术后肝再生过程中,使用γ分泌酶抑制剂Fli-06抑制Notch信号通路后,大鼠的肝再生能力明显降低,Notch-Hif-1α信号通路可能参与调控了大鼠肝部分切除术后肝再生过程。Objective： To investigate whether gamma secretase inhibitor Fli-06 could have an impact on the liver regeneration after liver resection in a rat model. With this in vivo model, we explore the underlying mechanism of notch signal pathway. Methods： SD rats were divided into control group（with intraperitoneal injection of normal saline, n=24） and experimental group（with intraperitoneal injection of gamma secretase inhibitor, n=24）. The two groups were treated by partial hepatectomy, and each group was divided into four subgroups according to the time point when remnant liver tissues were preserved, namely, 0 d, 1 d, 3 d and 5 d. The liver weight / body weight ratio was detected, the expression of PCNA in regenerating liver was detected by immunohistochemistry method, and the Notch1,Hes1 and VEGF m RNA in remnant liver tissues was detected by RT-PCR method. NICD and Hif-1 protein of the liver tissues during the liver regeneration process were analyzed and compared by Western-Blot methods. Results： 1. The liver weight/body weight ratio in inhibitor group was significantly lower than the control group. There was a significant difference between the two groups（P〈0.05）. 2. Immunohistochemical results suggested that the percentage of PCNA positive cells in the regenerating liver in the inhibitor group after 1 d, 3d, 5 d were significantly lower than the control group（P〈0.05）. 3. Western Blot results suggested that the expression levels of NICD and Hif-1 alpha in 1 d, 3 d,5 d were significantly lower than the control group（P〈0.05）. RT-PCR results indicated that Hes1 m RNA in inhibitor group were lower than the control group after 1 d, 3 d（P〈0.05）. VEGF m RNA in inhibitor group were lower than the control group after 3 d, 5 d（P〈0.05）. Conclusion： The ability of the liver regeneration of rats after hepatectomy decreased significantly after gamma secretase inhibitors inhibited Notch signaling pathway. The results showed that the Notch-Hif-1 alpha signaling pathway may be involved

关 键 词：肝再生 γ分泌酶抑制剂 NICD HIF-1Α 

分 类 号：Q95-3[生物学—动物学] R657.3[医药卫生—外科学]