PMID- 28073413 OWN - NLM STAT- MEDLINE DCOM- 20170928 LR - 20181202 IS - 1007-3418 (Print) IS - 1007-3418 (Linking) VI - 24 IP - 12 DP - 2016 Dec 20 TI - [Cellular and molecular mechanisms of anti-inflammatory effect of peroxisome proliferator-activated receptor alpha]. PG - 916-920 LID - 10.3760/cma.j.issn.1007-3418.2016.12.008 [doi] AB - Objective: To investigate the cellular and molecular mechanisms of the anti-inflammatory effect of peroxisome proliferator-activated receptor alpha (PPARalpha). Methods: Firstly, bone marrow-derived macrophages (BMDMs) were randomly divided into control group, LPS group, WY14643 10 mumol/L group, WY14643 25 mumol/L group, and WY14643 50 mumol/L group using a random number table. Secondly, BMDMs were randomly divided 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 PPARalpha WY14643 was administered at doses of 10, 25, and 50 mumol/L (50 mumol/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 cells 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-alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and mRNA expression of chemokine-1 (CXCL-1) and chemokine-10 (CXCL-10). Western blot was used to measure PPARalpha 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, PPARalpha 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-alpha, IL-1beta, IL-6, CXCL-1, and CXCL-10 was as follows in the control group, LPS group, WY14643 10 mumol group, WY14643 25 mumol group, and WY14643 50 mumol group: TNF-alpha (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-1beta (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, PPARalpha activation promoted autophagy in vitro in a dose-dependent manner. The results of Western blot and fluorescence microscopy in the control group, LPS group, WY14643 10 mumol group, WY14643 25 mumol group, and WY14643 50 mumol group showed that the expression of autophagy-related proteins and autophagosome formation gradually increased with the increasing concentration of WY14643. In vitro, WY14643 inhibited autophagy, promoted inflammatory response in primary macrophages, and reversed the anti-inflammatory effect of PPARalpha. The results of gene expression showed that the relative expression of TNF-alpha, IL-1beta, IL-6, CXCL-1, and CXCL-10 was as follows in the LPS group, WY14643+LPS group, and 3-MA+WY14643+LPS group: TNFalpha (4.327+/-0.478, 1.218+/-0.424, and 3.901+/-0.447, P < 0.05), IL-1beta (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: PPARalpha can promote cell autophagy and inhibit inflammatory response and may become a new therapeutic target for clinical prevention and treatment of inflammatory disease. FAU - Jiao, M J AU - Jiao MJ AD - Department of Infectious Diseases, Third Affiliated Hospital, Hebei Medical University, Shijiazhuang 050051,China. FAU - Zhou, L AU - Zhou L AD - Beijing Artificial Liver Treatment and Training Center, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China. FAU - Ren, F AU - Ren F AD - Beijing Institute of Hepatology, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China. FAU - Wang, Y D AU - Wang YD AD - Department of Infectious Diseases, Third Affiliated Hospital, Hebei Medical University, Shijiazhuang 050051,China. FAU - Shen, C AU - Shen C AD - Department of Infectious Diseases, Third Affiliated Hospital, Hebei Medical University, Shijiazhuang 050051,China. FAU - Duan, Z P AU - Duan ZP AD - Beijing Artificial Liver Treatment and Training Center, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China. FAU - Zhao, C Y AU - Zhao CY AD - Department of Infectious Diseases, Third Affiliated Hospital, Hebei Medical University, Shijiazhuang 050051,China. LA - chi PT - Comparative Study PT - Journal Article PL - China TA - Zhonghua Gan Zang Bing Za Zhi JT - Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology JID - 9710009 RN - 0 (Anti-Inflammatory Agents) RN - 0 (IL6 protein, human) RN - 0 (Interleukin-1beta) RN - 0 (Interleukin-6) RN - 0 (Lipopolysaccharides) RN - 0 (PPAR alpha) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Animals MH - *Anti-Inflammatory Agents MH - Gene Expression MH - Inflammation/*prevention & control MH - Interleukin-1beta MH - Interleukin-6 MH - *Lipopolysaccharides MH - Macrophages MH - PPAR alpha/*pharmacology MH - Random Allocation MH - Real-Time Polymerase Chain Reaction MH - Tumor Necrosis Factor-alpha EDAT- 2017/01/12 06:00 MHDA- 2017/09/29 06:00 CRDT- 2017/01/12 06:00 PHST- 2017/01/12 06:00 [entrez] PHST- 2017/01/12 06:00 [pubmed] PHST- 2017/09/29 06:00 [medline] AID - 10.3760/cma.j.issn.1007-3418.2016.12.008 [doi] PST - ppublish SO - Zhonghua Gan Zang Bing Za Zhi. 2016 Dec 20;24(12):916-920. doi: 10.3760/cma.j.issn.1007-3418.2016.12.008.