Notch2基因在大鼠肝移植术后胆管损伤中的表达及其意义

目的 探讨大鼠肝移植术后损伤胆管中Notch 2基因的表达及其意义.方法 将96只健康雄性SD大鼠随机分为3组,A组:未手术组;B组:假手术组;C组:采用改良"二袖套"法建立大鼠肝移植.于术后1、7、14、28 d检测总胆红素(TBIL)、γ-GT、谷丙转氨酶(ALT),同时处死大鼠取肝组织,苏木素-伊红(HE)染色观察病理改变,免疫组织化学、Western blot检测Notch 2基因的表达.结果 大鼠肝移植术后TBIL(32.05±3.28)μmol/L,γ-GT(118.30±17.40)μmol/L明显升高(P＜0.05),HE染色显示损伤胆管增生(14 d),部分胆管破坏、消失(28 d).免疫组织化学显示,A和B组胆管细胞Notch 2染色阴性,C组胆管细胞Notch 2染色强阳性,Western blot检测显示Notch 2蛋白在C组肝组织中呈逐渐升高的强表达(P＜0.05).结论 大鼠肝移植术后随胆管损伤加重,Notch 2基因表达显著升高,该基因可能参与了胆管损伤.

Abstract：

Objective To investigate the expression and clinical significance of Notch 2 expression in postoperative injured bile duct of rat liver transplantation. Methods Ninety-six healthy male SD rats were divided into three groups randomly: un-operation group (A), Sham operation group ( B), and group of liver transplantation model by modified "two-muff" techniques (C). Total bilirubin (TBIL), γ-GT and alanine aminotransferase (ALT) were determined at 1st, 7th, 14th, 28th day after transplantation and the expression of Notch 2 in the liver was detected by immunohistochemisty and Western blotting at the same time. Results TBIL (32. 05 ±3. 28) μmol/L, and γ-GT ( 118.30 ± 17.40) μmol/L were increased obviously after liver transplantation ( P ＜ 0. 05 ). Hematoxylin-eosin ( HE ) staining showed the injured bile ducts obviously proliferated after 14 days, and partly posttransplanted bile ducts were destructed and vanished after 28 days. Immunohistochemisty revealed that Notch 2 staining in bile ducts was negative in groups A and B, but Notch 2 staining in hyperplasitic bile ducts was strong positive in group C. Western blotting confirmed that Notch 2 protein was upregulated gradually in the damaged bile ducts after rat liver transplantation (P ＜ 0. 05 ). Conclusion Bile duct injury is aggravated after rat liver transplantation, and Notch 2 gene expression is up-regulated remarkably.

Expression and significance of Notch 2 gene in postoperative bile duct of rat liver transplantation

Objective To investigate the expression and clinical significance of Notch 2 expression in postoperative injured bile duct of rat liver transplantation. Methods Ninety-six healthy male SD rats were divided into three groups randomly: un-operation group (A), Sham operation group ( B), and group of liver transplantation model by modified "two-muff" techniques (C). Total bilirubin (TBIL), γ-GT and alanine aminotransferase (ALT) were determined at 1st, 7th, 14th, 28th day after transplantation and the expression of Notch 2 in the liver was detected by immunohistochemisty and Western blotting at the same time. Results TBIL (32. 05 ±3. 28) μmol/L, and γ-GT ( 118.30 ± 17.40) μmol/L were increased obviously after liver transplantation ( P ＜ 0. 05 ). Hematoxylin-eosin ( HE ) staining showed the injured bile ducts obviously proliferated after 14 days, and partly posttransplanted bile ducts were destructed and vanished after 28 days. Immunohistochemisty revealed that Notch 2 staining in bile ducts was negative in groups A and B, but Notch 2 staining in hyperplasitic bile ducts was strong positive in group C. Western blotting confirmed that Notch 2 protein was upregulated gradually in the damaged bile ducts after rat liver transplantation (P ＜ 0. 05 ). Conclusion Bile duct injury is aggravated after rat liver transplantation, and Notch 2 gene expression is up-regulated remarkably.