受体相互作用蛋白1在异烟肼致小鼠肝细胞坏死中的作用研究  

Effects of receptor interacting protein(RIP)1 on isoniazid induced hepatocyte necroptosis in mice

作　　者：赵宏宇[1] 胡晓[1] 沈海涛[1] 郑强[1] 

机构地区：[1]中国医科大学附属盛京医院急诊科,110004

出　　处：《天津医药》2016年第6期704-707,805,共4页Tianjin Medical Journal

基　　金：卫生部国家临床重点专科建设项目(2012-649);中国医科大学附属盛京医院课题资助项目(MC82)

摘　　要：目的研究异烟肼(INH)致小鼠肝细胞坏死是否与受体相互作用蛋白1(RIP1)的表达和程序性坏死有关。方法成年雄性昆明鼠18只按随机数字表法均分为3组,对照(C)组腹腔注射0.3 m L生理盐水每天1次;INH组以INH溶于生理盐水中,按100 mg/kg每天1次腹腔注射;Nec-1+INH组以Nec-1溶于0.1%二甲基亚砜(DMSO),1 mg/kg腹腔每12 h注射1次,同时INH腹腔注射,剂量同INH组。所有动物均被干预7 d。HE染色观察细胞形态学变化。免疫组化、免疫印迹法和实时荧光定量PCR法检测各组RIP1的表达。比色法检测各组丙二醛(MDA)、活性氧(ROS)、谷胱甘肽(GSH)和超氧化物歧化酶(SOD)的含量。结果 C组的肝细胞排列整齐,Nec-1+INH组肝细胞存在变性和坏死,但程度较INH组明显减轻。与C组比较,INH组肝细胞RIP1、ROS和MDA表达均增强,GSH和SOD表达下降(P<0.05)。与INH组比较,Nec-1+INH组肝细胞坏死明显减轻,肝细胞RIP1、MDA和ROS的表达明显下降,GSH和SOD的表达增加(P<0.05)。结论 INH致小鼠肝细胞坏死与程序性细胞坏死有关,与RIP1表达增强有关。阻断RIP1的表达,可能是一个预防INH致肝细胞坏死的策略。Objective To study the relationship between receptor interacting protein (RIP)1 and hepatocyte necropto?sis in isoniazid (INH) induced mouse model. Methods Kunming male mice were randomly divided into three groups. Con?trol group (C) received 0.3 mL of normal saline one time per day. INH group (INH) was injected intraperitoneally INH 100 mg/kg body weight, one time per day. Nec-1+INH group was injected intraperitoneally Nec-1 in 0.1%DMSO and 1 mg/kg body weight one time/12 hours, and INH was injected intraperitoneally at the same dose with that of INH group. All animals were treated for 7 days. Pathological changes of liver tissues were studied by HE staining. RIP1 expression was detected by immunohistochemical, Western blot and real-time PCR analysis. Levels of malondialdehyde (MDA), reactive oxygen species (ROS), glutathione (GSH) and superoxide dismutase (SOD) in liver homogenate were determined by colorimetric method. Re?sults Hepatocytes were arranged orderly in C group. The degeneration and necrosis of hepatocytes were found in Nec-1+INH group, and severe degeneration and necrosis of hepatocytes were found in INH group. Compared with C group, the ex?pression levels of RIP1, ROS and MDA were increased significantly, and the expression levels of GSH and SOD were de?creased significantly in INH group (P<0.05). INH-induced acute liver necroptosis was significantly alleviated after treat?ment with Nec-1. Compared with INH group, the expression levels of RIP1, MDA and ROS were significantly decreased, and the expression levels of GSH and SOD were significantly increased in Nec-1+INH group (P<0.05). Conclusion These re?sults suggest that RIP1 is involved in INH-induced hepatocyte necroptosis in mice. The inhibition of RIP1 expression might be a treatment strategy for prohibition of INH-induced acute liver necroptosis.

关 键 词：异烟肼 肝细胞 程序性坏死 受体相互作用蛋白1 

分 类 号：R575.32[医药卫生—消化系统]