TIPE2在小鼠巨噬细胞焦亡中的作用  

Role of TIPE2in macrophage pyroptosis in mice

作　　者：孔倩[1] 段薇娜[1] 李文澜[1] 江梦[1] 邱珍[1] 黄琴[1] 李文远 吴晓静[1] Kong Qian;Duan Weina;Li Wenlan;Jiang Meng;Qiu Zhen;Huang Qin;Li Wenyuan;Wu Xiaojing(Department of Anesthesiology,Renmin Hospital,Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,430060

出　　处：《中华麻醉学杂志》2018年第8期1001-1004,共4页Chinese Journal of Anesthesiology

基　　金：湖北省自然科学基金(2016CFB251).

摘　　要：目的 评价肿瘤坏死因子-α诱导蛋白家族8-样蛋白2(TIPE2)在小鼠巨噬细胞焦亡中的作用.方法 采用随机数字表法将J774A.1巨噬细胞分为4组(n=18):空载对照组(C组)、空载对照组+LPS∕ATP组(C+LPS∕ATP组)、TIPE2过表达组(T组)、TIPE2过表达+LPS∕ATP组(T+LPS∕ATP组).C组和C+LPS∕ATP组用不携带TIPE2的慢病毒感染细胞,T组和T+LPS∕ATP组构建TIPE2过表达稳转细胞系,随后C+LPS∕ATP组和T+LPS∕ATP组加入LPS 1.0μg∕ml,孵育5h后再加入ATP 5.0 mmol∕L孵育1 h.收集细胞,采用Western blot法检测TIPE2、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、IL-1β和IL-18的表达;采用流式细胞术检测焦亡细胞百分比;采用ELISA法检测细胞培养液TNF-α、IL-6、IL-1β和IL-18的浓度.结果 与C组比较,C+LPS∕ATP组TIPE2表达下调,NLRP3、IL-1β和IL-18表达上调,焦亡细胞百分比、培养液TNF-α、IL-6、IL-1β和IL-18浓度升高(P<0.05).与T组比较,T+LPS∕ATP组TIPE2表达下调,NLRP3、IL-1β和IL-18表达上调,焦亡细胞百分比、培养液TNF-α、IL-6、IL-1β和IL-18浓度升高(P<0.05).与C+LPS∕ATP组比较,T+LPS∕ATP组TIPE2表达上调,NLRP3、IL-1β和IL-18表达下调,焦亡细胞百分比、培养液TNF-α、IL-6、IL-1β和IL-18浓度降低(P<0.05).结论 TIPE2可抑制小鼠巨噬细胞焦亡,机制可能与抑制NLRP3炎症小体激活有关.Objective To evaluate the role of tumor necrosis factor alpha-inducible protein 8 like-2 ( TIPE2) in macrophage pyroptosis in mice. Methods Mouse macrophages J774A. 1 were seeded in 6-cm culture dishes (5 ml∕dish) at the density of 2×10^5 cells∕ml and divided into 4 groups (n=18 each) using a random number table method: blank vector control group ( C group) , blank vector plus lipopolysaccharide ( LPS)∕ATP group ( C+LPS∕ATP group) , TIPE2 overexpression group ( T group) and TIPE2 overexpres-sion plus LPS∕ATP group ( T+LPS∕ATP group) . Cells were infected with lentivirus without TIPE2 in C and C+LPS∕ATP groups. TIPE2 overexpression stable cell line was constructed in T group and T+LPS∕ATP group. LPS 1. 0 μg∕ml was added and cells were incubated for 5 h, and then ATP 5. 0 mmol∕L was added and cells were incubated for 1 h in C+LPS∕ATP group and T+LPS∕ATP group. Cells were collected for de-tection of the expression of TIPE2, NOD-like receptor protein 3 ( NLRP3) , interleukin-1beta ( IL-1β) and interleukin-8 ( IL-18) by Western blot. Flow cytometry was used to detect the pyroptotic cells, and the per-centage of pyroptotic cells was calculated. The concentrations of tumor necrosis factor-alpha ( TNF-α) , IL-6, IL-1β and IL-18 in cell culture media were determined by enzyme-linked immunosorbent assay. Results Compared with group C, the expression of TIPE2 was significantly down-regulated, the expression of NL-RP3, IL-1β and IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group C+LPS∕ATP (P<0. 05). Com-pared with group T, the expression of TIPE2 was significantly down-regulated, the expression of NLRP3, IL-1βand IL-18 was up-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were increased in group T+LPS∕ATP ( P<0. 05) . Compared with group C+LPS∕ATP, the expression of TIPE2 was significantly up-regulated,

关 键 词：细胞死亡 TIPE2 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]