民猪脂肪细胞分化过程中抗寒相关基因表达规律研究

旨在探究民猪脂肪细胞分化过程中产热基因和米色脂肪标记基因的表达变化规律，为进一步研究民猪脂肪细胞向米色脂肪细胞分化的分子机制提供依据。本研究采集1月龄民猪的背部脂肪组织，分离前体脂肪细胞进行培养及成脂诱导，观察分化过程中细胞形态并进行油红O染色鉴定；检测产热基因UCP3、PGC-1α、PPARα和米色脂肪标记基因EBF2、CD81、PDGFRα在诱导分化第0、2、4、6、8天的表达量。结果表明，随着诱导分化的时间增加，细胞中的脂滴逐渐增多，在分化第8天时进行油红O染色，多数细胞达到成熟脂肪细胞阶段。以第0天为对照，PGC-1α、EBF2、PDGFRα在第2天时表达量极显著升高（P<0.01），且达到最高，第4、6、8天呈下降趋势。UCP3在第4、6、8天表达量极显著升高（P<0.01），第8天有所下降。PPARα和CD81在第2天时表达量极显著升高（P<0.01），且在第4和6天表达量与第2天相比变化不大。综上表明，本研究成功进行了民猪前体脂肪细胞的成脂诱导分化，揭示了分化过程中产热和米色脂肪标记基因的表达规律，为进一步研究民猪前体脂肪细胞向米色脂肪细胞分化的分子机制和民猪抗寒能力奠定基础。

Expression of Cold Resistance Related Genes during Differentiation of Adipocytes in Min Pig

This study aimed to investigate the expression of thermogenic genes and beige fat marker genes during the differentiation of Min pig adipocytes and to provide a basis for further study on the molecular mechanism of differentiation of Min pig adipocytes into beige adipocytes. Adipose tissue of 1-month-old Min pig was collected, preadipocytes were isolated, cultured and induced to adipogenesis, cell morphology was observed during differentiation and identified by Oil red O staining; The expression levels of thermogenic genes UCP3, PGC-1α, PPARα, and beige fat marker genes EBF2, CD81, PDGFRα were detected on day 0, 2, 4, 6 and 8 after differentiation. The results showed that the number of lipid droplets increased with the time of differentiation increased. The result of Oil red O staining showed that most of the cells reached the stage of mature adipocytes. The expression levels of PGC-1α, EBF2 and PDGFRα were significantly increased on day 2 compared with those on day 0 (P<0.01) and reached the highest level, and decreased on day 4, 6, and 8. The expression of UCP3 was significantly increased at day 4, 6, and 8(P<0.01), and decreased at day 8. The expression levels of PPARα and CD81 were significantly increased on day 2 (P<0.01), and there was no significant change on day 4, 6 compared with day 2. In conclusion, this study successfully conducted adipogenic differentiation of Min pig preadipocytes, revealed the expression of thermogenic and beige fat marker genes during differentiation, and laid a foundation for further study on the molecular mechanism of differentiation of Min pig preadipocytes into beige adipocytes and cold-resistance of Min pig.