Metformin regulate the balance of Th17/regulatory T cells and expression of serum cytokines through AMPK-mTOR signaling pathway in collagen-induced arthritis
10.3760/cma.j.issn.1007-7480.2016.09.008
- VernacularTitle:二甲双胍通过腺苷酸活化蛋白酶-哺乳动物雷帕霉素靶蛋白信号通路调节胶原诱导性关节炎大鼠辅助性T细胞17/调节性T细胞的分化及细胞因子的表达
- Author:
Mingfeng YANG
;
Yu DING
;
Yiwen WANG
;
Bin ZHANG
;
Hongzhi WANG
- Publication Type:Journal Article
- Keywords:
Metformin;
Arthritis,experimental;
T-lymphocytes,regulatory;
T-lymphocytes,helper-inducer;
AMPK-mTOR
- From:
Chinese Journal of Rheumatology
2016;20(9):614-618
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate therapeutic effects of metformin on the expression of serum cytokines,balance of splenic Th17/regulatory T cells (Treg) and expression of splenic AMPK-mTOR in collagen-induced arthritis (CIA) rats,and the mechanism thereof.Methods The rat model of CIA was established by injecting bovine type Ⅱ collagen.Forty rats were randomly divided into five groups:the CIA model group(sterile water by gavage),Met-30 mg/kg group (metformin 30 mg ·kg-1 ·d-1 by gavage),Met100 mg/kg group(metformin 100 mg ·kg-1 ·d-1 by gavage),Met-300 mg/kg group(metformin 300 mg ·kg-1 ·d-1 by gavage) and control group (sterile water by gavage).The hind paw volume was recorded once a week.The serum level of cytokines,tumor necrosis factor (TNF)-α,interleukin (IL)-1β,IL-6,and IL-17,were measured by enzyme linked immunosorbent assay (ELISA) 35 days after the initial immunization.The quantity of Th17 and Treg cells were examined by flow cytometry.The expression of AMPK,p-AMPK,mTOR and p-mTOR were examined by western blotting.One-way analysis of variance was used to evaluate the experimental data.Results The values of hind paw volume were decreased on the 35 d of the initial immunization in the Met100 mg/kg [(2.43±0.37) ml,t=2.97,P＜0.05] and Met-300 mg/kg groups [(2.58±0.21) ml,t=2.96,P＜0.05] than those of CIA model group (2.97±0.23) ml.The serum levels of TNF-α,IL-1β,IL-6 and IL-17 on the 35-d after the initial immunization were significantly lower in the metformin therapeutic groups [Met-300 mg/kg group TNF-α (104±8) pg/ml,t=42.77,P＜0.05;IL-1β (183±24) pg/ml,t=60.457,P＜0.05;IL-6 (19.3±2.8) pg/ml,t=53.62,P＜0.05;IL-17 (64.5±6.7) pg/ml,t=47.92,P＜0.05] than those of the CIA model group [TNF-α (246±8) pg/ml;IL-1β (1 336±40) pg/ml;IL-6 (87.0±5.1) pg/ml;IL-17 (282.3±6.8) pg/ml].The quantity of Th17 and Treg cells were significantly different in the Met-300 mg/kg group than those of the CIA model group 35 d after the initial immunization [Th17(6.57±0.39) vs (9.89±0.53),t=8.74,P＜0.05;Treg (7.60±0.45) vs (3.94±0.61),t =8.37,P＜0.05].The expression of p-AMPK on the 35 d after the initial immunization in the metformin therapeutic groups was significantly higher than in the CIA model group(P＜0.05),and the expression of p-mTOR was significantly lower (P＜0.05).Conclusion Metformin can significantly inhibit the serum levels of TNF-α,IL-1β,IL-6 and IL-17 in CIA model rats,and regulate the balance of Th17/Treg through AMPK-mTOR signaling pathway.
