过表达UCP3基因对萨湖羊前体脂肪细胞分化的影响

doi:10.11843/j.issn.0366-6964.2023.08.014

Abstract

Abstract: The purpose of this study was to clarify the tissue and cell expression pattern of uncoupling protein 3 (UCP3) gene in Sahu sheep, and to clarify the effect of overexpression of UCP3 on revealing the differentiation of preadipocytes in Sahu sheep. This study used real-time quantitative PCR (qPCR) technology to detect the expression distribution of UCP3 gene in the heart, liver, spleen and other tissues of 3 Sahu sheep, as well as its expression levels at different stages of adipogenic differentiation; After overexpression of UCP3, qPCR was used to verify the overexpression efficiency, using Oil Red O staining, lipid extraction analysis, and adipocyte differentiation marker genes testing, clarify the effects of overexpression of UCP3 on lipid production in Sahu sheep preadipocytes from both cellular morphology and biological perspectives.The results showed that the UCP3 gene was widely expressed in various tissues of Sahu sheep, with relatively higher expression levels in the heart and spleen; The temporal expression profile showed that the expression level of UCP3 gene showed an upward trend during the differentiation of preadipocytes in Sahu sheep (P<0.05); After overexpression of UCP3, the number of lipid droplets in the preadipocytes of Sahu sheep increased significantly (P<0.01); In addition, the mRNA levels of PPAR γ, C/EBP α and LEP were extremely significantly up-regulated (P<0.01), and FAS and UCP1 mRNA levels were significantly up-regulated (P<0.05). This study revealed the expression pattern of UCP3 in Sahu sheep and verified that overexpression of UCP3 can promote adipogenic differentiation of Sahu sheep preadipocytes. It is speculated that UCP3 is a positive regulatory factor of Sahu sheep preadipocytes, it possibly regulates the differentiation of preadipocytes through regulating the expression of PPAR γ, C/EBPα and FAS genes.

Key words: Sahu sheep, preadipocytes, pcDNA3.1-flag, UCP3, seamless cloning

CLC Number:

S826.2

LIU Linli, PENG Xuelan, LI Bo, CHENG Lefan, CIREN Lamu, ZHANG Enping. Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3275-3285.

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Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes

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