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Peroxisome proliferator-activated receptor ? isoforms differentially regulate preadipocyte proliferation, apoptosis, and differentiation in chickens.

ABSTRACT: Peroxisome proliferator-activated receptor ? (PPAR?) has 2 protein isoforms (PPAR?1 and PPAR?2) generated by alternative promoter usage and alternative splicing. However, their functional uniqueness and similarity remain unclear. In the study, we investigated the effects of lentivirus-mediated overexpression of PPAR?1 and PPAR?2 on proliferation, apoptosis, and differentiation of the immortalized chicken preadipocytes. Cell Counting Kit-8 assay showed PPAR?1 and PPAR?2 overexpression markedly suppressed cell proliferation, and fluorescence activated cell sorting analysis showed that PPAR?1 and PPAR?2 overexpression caused cell cycle arrest at G0/G1 phase. Cell death detection ELISA analysis showed both PPAR?1 and PPAR?2 overexpression induced cell apoptosis. Oil red O staining and gene expression analysis showed both PPAR?1 and PPAR?2 overexpression promoted preadipocyte differentiation. In the presence of PPAR? ligand, rosiglitazone, PPAR?2 overexpression was more potent in inducing apoptosis, promoting adipogenesis, and suppressing cell proliferation than PPAR?1 overexpression. We further explored the molecular basis for their functional differences. Reporter gene assay showed that under ligand conditions, PPAR?2 overexpression resulted in 1.68-fold increase in transcription activity compared with PPAR?1. Electrophoretic mobility shift assay showed both PPAR?1 and PPAR?2 could bind to PPAR response element (PPRE) as heterodimer with retinoid X receptor alpha, and by comparison, PPAR?2 had a higher affinity for PPRE than PPAR?1. Reporter gene assay showed expression PPAR?1 and PPAR?2 similarly induced fatty acid synthase and adipocyte fatty acid-binding protein promoter activity but differentially induced lipoprotein lipase and perilipin 1 promoter activities. Coimmunoprecipitation analysis showed that PPAR?1 and PPAR?2 interacted similarly with the coactivators, Tat-interacting protein 60. Taken together, our results demonstrate that PPAR?1 and PPAR?2 differentially regulate preadipocyte proliferation, apoptosis, and differentiation as a result of their distinct and overlapping molecular functions.

SUBMITTER: Mu F

PROVIDER: S-EPMC7705046 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Peroxisome proliferator-activated receptor γ isoforms differentially regulate preadipocyte proliferation, apoptosis, and differentiation in chickens.

Mu Fang F Jing Yang Y Ning Bolin B Huang Jiaxin J Cui Tingting T Guo Yaqi Y You Xin X Yan Xiaohong X Li Hui H Wang Ning N

Poultry science 20201008 12

Peroxisome proliferator-activated receptor γ (PPARγ) has 2 protein isoforms (PPARγ1 and PPARγ2) generated by alternative promoter usage and alternative splicing. However, their functional uniqueness and similarity remain unclear. In the study, we investigated the effects of lentivirus-mediated overexpression of PPARγ1 and PPARγ2 on proliferation, apoptosis, and differentiation of the immortalized chicken preadipocytes. Cell Counting Kit-8 assay showed PPARγ1 and PPARγ2 overexpression markedly su ...[more]

PMID: 33248556

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Project description:Ascites (serous fluid accumulation in the abdominal cavity) has been observed worldwide in fast growing broilers. Pulmonary vascular remodeling is an important pathological feature of broiler ascites syndrome. Peroxisome proliferators-activated receptor gamma (PPARγ) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) are expressed in pulmonary vascular endothelial cells and vascular smooth muscle cells (VSMC) where they participate in the regulation of normal pulmonary vascular function. The objective of the present study was to investigate the effects of omega-3 fatty acids (found in fish oil) and pioglitazone (PIO) as natural and synthetic PPARγ ligands supplementation on PPARγ and PGC-1α expression in the prevention of pulmonary arterial hypertension (PAH) syndrome in broiler chickens. The experiment was conducted with 4 treatment groups: 1) negative control, normal temperature conditions with basal diet; 2) positive control, low-temperature conditions with basal diet; 3) positive control + 10 mg PIO/kg of weight/d and 4) positive control + 1% FO. Each treatment had 5 replicates. Ascites heart index (RV/TV) was significantly (P < 0.05) reduced in chickens receiving FO (0.20) and PIO (0.21) compared to the positive control group (0.26). The addition of PIO in broilers under cold-induced ascites significantly increased the expression of PPARγ (9.44) and PGC-1α (5.81) genes in lung tissue compared to the negative control group (1.03, P < 0.05). Proliferative indexes of VSMC in pulmonary arteries such as PMT, PIT, and percentage wall thickness were significantly elevated in positive control group, indicating that pulmonary vascular remodeling occurred following VSMC proliferation in ascites. The vessel internal diameter was increased in FO and PIO groups. Based on these results, activation and expression of PPARγ and PGC-1α genes as a critical regulator of pulmonary artery smooth muscle cell using ligands, especially PIO, can be effective in reducing the incidence of PAH in broiler chickens.

Dataset Information

Peroxisome proliferator-activated receptor ? isoforms differentially regulate preadipocyte proliferation, apoptosis, and differentiation in chickens.

Publications

Peroxisome proliferator-activated receptor γ isoforms differentially regulate preadipocyte proliferation, apoptosis, and differentiation in chickens.

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