Role of TIPE2 in macrophage pyroptosis in mice
10.3760/cma.j.issn.0254-1416.2018.08.027
- VernacularTitle:TIPE2在小鼠巨噬细胞焦亡中的作用
- Author:
Qian KONG
1
;
Weina DUAN
;
Wenlan LI
;
Meng JIANG
;
Zhen QIU
;
Qin HUANG
;
Wenyuan LI
;
Xiaojing WU
Author Information
1. 430060,武汉大学人民医院麻醉科
- Keywords:
Cell death;
TIPE2
- From:
Chinese Journal of Anesthesiology
2018;38(8):1001-1004
- CountryChina
- Language:Chinese
-
Abstract:
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 2×105 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, the expression of NLRP3, IL-1β and IL-18 was down-regulated, and the percentage of pyroptotic cells and concentrations of TNF-α, IL-6, IL-1β and IL-18 in cell culture media were decreased in group T+LPS∕ATP ( P<0. 05) . Conclusion TIPE2 can inhibit macrophage pyroptosis, and the mechanism may be related to inhibiting activation of NL-RP3 inflammasome in mice.
