Cellular and molecular mechanisms of anti-inflammatory effect of peroxisome proliferator-activated receptor α
10.3760/cma.j.issn.1007-3418.2016.12.008
- VernacularTitle:过氧化物酶体增殖物激活受体α抗炎作用的细胞与分子机制研究
- Author:
Mingjing JIAO
1
;
Li ZHOU
;
Feng REN
;
Yadong WANG
;
Chuan SHEN
;
Zhongping DUAN
;
Caiyan ZHAO
Author Information
1. 河北医科大学第三医院感染科
- Keywords:
Peroxisome proliferator-activated receptors;
Autophagy;
Inflammation
- From:
Chinese Journal of Hepatology
2016;24(12):916-920
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the cellular and molecular mechanisms of the anti-inflammatory effect of peroxisome proliferator-activated receptor α (PPARαt).Methods Firstly,bone marrow-derived macrophages (BMDMs) were randomly divided into control group,LPS group,WY14643 10 μmol/L group,WY14643 25 μmol/L group,and WY14643 50 μmol/L group using a random number table.Secondly,BMDMs were randomly dividcd into LPS group,WY14643+LPS group,and 3-MA+WY14643+LPS group.Primary BMDMs were stimulated by LPS (20 ng/ml) to establish the cellular model of inflammation.The selective agonist of PPARα WY14643 was administered at doses of 10,25,and 50 μmol/L (50 μmol/L for the second part of the experiment) at 2 hours before model establishment.The autophagy inhibitor 3-MA was administered at a dose of 10 mmol/L at 2 hours before model establishment.The cells in the control group were treated with dimethylsulfoxide (DMSO) at the same dose.The calls were transfected with GFP-LC3 plasmids at 24 hours before model establishment.The cells were harvested at 6 hours after LPS stimulation and related tests were performed.Green fluorescent protein was measured under a fluorescence microscope to evaluate autophagy activity.Quantitative real-time PCR was used to measure tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β),interleukin-6 (IL-6),and mRNA expression of chemokine-1 (CXCL-1) and chemokine-10 (CXCL-10).Westem blot was used to measure PPARα and autophagy-related proteins LC3,ATG-5,ATG-7,and LAMP-1.A one-way analysis of variance was used for comparison between groups,and the LSD-t test was used for comparison between any two groups.Results In vitro,PPARα activation inhibited LPS-induced inflammatory response in primary macrophages in a dose-dependent manner.The results of gene expression showed that the relative expression of TNF-α,IL-1β,IL-6,CXCL-1,and CXCL-10 was as follows in the control group,LPS group,WY14643 10 μmol group,WY14643 25 μmol group,and WY14643 50 μmol group:TNF-α (0.085±0.009,4.065±0.544,3.281±0.368,1.780±±0.293,and 0.781±0.303,P ＜ 0.01),IL-1β (0.081±0.017,0.776±0.303,0.225±0.154,0.161±0.068,and 0.101±0.025,P ＜ 0.05),IL-6 (0.041±0.011,0.189±0.014,0.144±0.033,0.126±0.013,and 0.048±0.015,P ＜ 0.01),CXCL-1 (0.051±0.011,0.515±0.145,0.356±0.078,0.257±0.068,and 0.069±0.030,P ＜ 0.01),and CXCL-10 (0.126±0.068,0.831±0.093,0.508±0245,0.474±0.047,and 0.204±0.021,P ＜ 0.05).In vitro,PPARα activation promoted autophagy in vitro in a dose-dependent manner.The results of Westem blot and fluorescence microscopy in the control group,LPS group,WY14643 10 μmol group,WY14643 25 μmol group,and WY14643 50 μmol group showed that the expression of autophagy-related proteins and autophagosome formation gradually increased with the increasing concentration of WY14643.In vitro,WY 14643 inhibited autophagy,promoted inflammatory response in primary macrophages,and reversed the anti-inflammatory effect of PPARα.The results of gene expression showed that the relative expression of TNF-α,IL-1β,IL-6,CXCL-1,and CXCL-10 was as follows in the LPS group,WY14643+LPS group,and 3-MA+WY14643+LPS group:TNFα (4.327±.478,1.218±0.424,and 3.901±0.447,P ＜ 0.05),1L-1β (4.277±0.407,1.418±0.424,and 3.029±0.192,P ＜ 0.01),IL-6 (4.175±0.549,1.373±0.499,and 4.031±0.475,P ＜ 0.05),CXCL-1 (8.199±1.149,2.024±0.547,and 5.973±0.843,P ＜ 0.05),and CXCL-10 (1.208±0.148,0.206±0.069,and 0.798±0.170,P ＜ 0.05).Conclusion PPARα can promote cell autophagy and inhibit inflammatory response and may become a new therapeutic target for clinical prevention and treatment of inflammatory disease.
