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The PPARγΔ5 Expression During Sus scrofa Adipocyte Differentiation and Its Effect on the Cell Differentiation |
WEI Fang1, JIANG Su-Su1,2, LI Shuai-Bing1, FU Lu1, SONG Su-Di1, LU Jian-Xiong1*, ZHANG Guo-Hua1* |
1 College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China; 2 Department of Animal Science and Technolo‐gy, Gansu Agriculture Technology College, Lanzhou 730000, China |
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Abstract Alternative splicing is an important way for eukaryotes to precisely regulate cell types and devel‐ opment. Peroxisome proliferator activated receptor γ (PPARγ) is a key regulator of adipocyte differentiation. Its pre-mRNA could produce a subtype of PPARγΔ5 through alternative splicing and affecting adipocyte dif‐ ferentiation. In this study, the preadipocytes were isolated from subcutaneous adipose tissue of three-day-old piglets (Sus scrofa) and induced to adipogenic differentiation. The mRNA expression of related genes was de‐tected by qPCR. it was observed that porcine adipocytes expressed PPARγΔ5 mRNA, and the expression in‐creased significantly in the early stage of differentiation, peaked in the middle and decreased in the later stage (P<0.01), which was consistent with the expressions of PPARγ and serine/arginine-rich splicing factor 1 (SRSF1). The expressions of PPARγΔ5 and SRSF1 in the adipocytes were extremely significant promoted by rosiglitazone treatment (P<0.01), and reduced by GW9662 (P<0.01). Furthermore, the PPARγΔ5-siRNA re‐ combinant lentivirus was constructed, and after transfected with porcine preadipocytes, the PPARγΔ5 expres‐ sion significantly decreased about 75% (P<0.01). Compared with the cells transfected with negative control lentivirus or non-transfected, PPARγΔ5 expression silencing significantly stimulated the differentiation of pre‐ adipocytes and the mRNA expressions of PPARγ, fatty acid binding protein 4 (FABP4) and stearoyl-CoA de‐ saturase (SCD) (P<0.05), and decreased the expression of lipoprotein lipase (LPL) (P<0.05). These results suggested that the porcine adipocytes expressed PPARγΔ5 through selective splicing depending on SRSF1 and the expression and activity of PPARγ. PPARγΔ5 was an inhibitor of adipocyte differentiation, which could at‐ tenuated the differentiation of porcine preadipocytes via negatively regulating PPARγ expression. This study provides a new clue for understanding the regulation of porcine adipogenesis.
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Received: 21 April 2022
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Corresponding Authors:
* lu2003jx@163.com; zhgh513@126.com
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