论文部分内容阅读
目的探讨静脉注射携带hIL-10腺病毒载体对淤胆性肝硬化大鼠肝常温缺血再灌注损伤的保护作用。方法结扎SD大鼠的胆总管建立淤胆性肝硬化模型,4周后通过每只尾静脉注射Ad-hIL10-EGFP1×109PFU/ml,体内转染72h后,进行常温肝缺血再灌注损伤实验(缺血15min,再灌注60min)。经静脉取血,进行肝功能生化检测,再通过酶联免疫吸附法(ELISA)检测大鼠血清内hIL-10的表达情况。并取肝脏冰冻切片,用倒置荧光显微镜观察肝组织中EGFP的表达,石蜡切片HE染色观察肝脏的组织形态变化;免疫组化方法观察肝脏内hIL-10的表达情况;Tunel法检测肝脏内细胞凋亡情况。结果治疗组的肝脏冰冻切片,荧光显微镜下可见EGFP的表达;同时免疫组化有较多的hIL-10染色;ELISA检测血清中hIL-10的表达量为(723·8±301·7)ng/ml。相对应的是,治疗组的肝功能较对照两组明显改善,组织病理改变较对照两组减轻,肝细胞凋亡数量明显减少,差异有显著意义(P<0·05)。结论hIL-10肝脏基因治疗可以减轻肝硬化大鼠的肝缺血再灌注损伤,其机制可能是减少肝缺血再灌注损伤过程中肝细胞凋亡的数量。
Objective To investigate the protective effect of intravenous injection of hIL-10 adenovirus on liver ischemia-reperfusion injury in cholestatic cirrhotic rats. Methods The model of cholestatic cirrhosis was established by ligating the common bile duct of SD rats. After 4 weeks, Ad-hIL10-EGFP1 × 109 PFU / ml was injected into each caudal vein and transfused for 72 hours in vivo. (Ischemia 15min, reperfusion 60min). The blood was taken from the venous and the biochemical tests of liver function were performed. The expression of hIL-10 in serum was detected by enzyme linked immunosorbent assay (ELISA). The liver slices were harvested, and the expression of EGFP in liver tissues was observed by inverted fluorescent microscope. The histological changes of liver were observed by HE staining. The expression of hIL-10 in liver was observed by immunohistochemical method. Death situation. Results The frozen sections of the liver of the treated group were observed under the fluorescence microscope, and the expression of EGFP was observed under the fluorescence microscope. At the same time, there was more hIL-10 staining in the immunohistochemistry. The level of hIL-10 in the serum was (723.8 ± 301.7) ng / ml. Correspondingly, the liver function of the treatment group was significantly improved compared with the control group. The histopathological changes were alleviated compared with the control group, and the number of hepatocyte apoptosis was significantly reduced (P <0.05). Conclusion The gene therapy of hIL-10 liver can reduce hepatic ischemia-reperfusion injury in cirrhotic rats, which may be the result of reducing hepatocellular apoptosis during liver ischemia-reperfusion injury.