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

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 mL 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。

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

Abstract: Objective To study the relationship between receptor interacting protein (RIP)1 and hepatocyte necroptosis in isoniazid (INH) induced mouse model. Methods Kunming male mice were randomly divided into three groups. Control 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. Results 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 expression levels of RIP1, ROS and MDA were increased significantly, and the expression levels of GSH and SOD were decreased significantly in INH group (P＜0.05). INH-induced acute liver necroptosis was significantly alleviated after treatment 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 results 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.