Shi Mingyue, Yang Shuai, Huang Xiaoyu, Wang Shouyuan, Li Wenxia, Yun Jiale, Lu Chang, Yang Yang, Cai Chunbo, Gao Pengfei, Guo Xiaohong, Li Bugao, Cao Guoqing
College of Animal Science, Shanxi Agricultural University, Taigu, China.
Anim Sci J. 2023 Jan-Dec;94(1):e13822. doi: 10.1111/asj.13822.
Fat deposition is one of the key factors affecting the economic development of pig husbandry. The aim of this study was to investigate the expression characteristics of caveolae-associated protein 3 (CAVIN3) and to elucidate its effect and mechanism on adipogenic differentiation of porcine preadipocytes. Cell transfection, quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot, and oil red O staining were used to detect the effect of CAVIN3 on the differentiation of porcine preadipocytes. The results showed that CAVIN3 was expressed in various tissues, with higher expression in adipose tissue, differentially expressed during cell adipogenic differentiation, and mainly distributed in the cytoplasm. Functional studies showed that, after CAVIN3 interference in preadipocytes, the expression of adipogenic factors and the content of lipid droplets were significantly decreased (p < 0.05). The results were reversed after CAVIN3 was overexpressed. The mechanism research showed that LY3214996 inhibited the extracellular signal-regulated kinase (ERK) phosphorylation and further inhibited lipogenic factors expression. Overexpression of CAVIN3 attenuates the inhibitory effect of LY3214996 on ERK phosphorylation and attenuates its inhibitory effect on adipogenic differentiation. Therefore, this study demonstrated that CAVIN3 promotes the differentiation of porcine preadipocytes by promoting ERK phosphorylation. The present study can lay a theoretical foundation for further studying the molecular mechanism of porcine fat deposition.
脂肪沉积是影响养猪业经济发展的关键因素之一。本研究旨在探讨小窝相关蛋白3(CAVIN3)的表达特征,并阐明其对猪前体脂肪细胞成脂分化的影响及机制。采用细胞转染、定量逆转录聚合酶链反应(qRT-PCR)、蛋白质免疫印迹法和油红O染色检测CAVIN3对猪前体脂肪细胞分化的影响。结果表明,CAVIN3在各组织中均有表达,在脂肪组织中表达较高,在细胞成脂分化过程中差异表达,主要分布于细胞质中。功能研究表明,前体脂肪细胞中CAVIN3干扰后,成脂因子表达及脂滴含量显著降低(p < 0.05)。CAVIN3过表达后结果相反。机制研究表明,LY3214996抑制细胞外信号调节激酶(ERK)磷酸化,进而抑制脂肪生成因子表达。CAVIN3过表达减弱LY3214996对ERK磷酸化的抑制作用,减弱其对成脂分化的抑制作用。因此,本研究表明CAVIN3通过促进ERK磷酸化促进猪前体脂肪细胞分化。本研究可为进一步研究猪脂肪沉积的分子机制奠定理论基础。