miR-128-1-5p对绵羊前体脂肪细胞增殖和分化调节作用的研究

doi:10.11843/j.issn.0366-6964.2020.005

Abstract

Abstract: This study aimed to reveal the regulating role of miR-128-1-5p in the proliferation and differentiation of ovine preadipocytes. The target genes of miR-128-1-5p were theoretically predicted and experimentally validated. After overexpressing miR-128-1-5p, the expression of proliferation marker genes were detected by qPCR, and the cell proliferation was examined by CCK-8 and EdU. The expression trends of miR-128-1-5p and its target genes during the differentiation of ovine preadipocytes were detected by qPCR and Western blotting. The regulatory mechanism of miR-128-1-5p on the ovine preadipocyte differentiation was revealed by overexpressing or inhibiting miR-128-1-5p. The cell adipogenic effect was tested by Oil Red O staining. The results showed that KLF2 was a target gene of miR-128-1-5p. During the proliferation of ovine preadipocytes, after miR-128-1-5p was overexpressed, the expressions of proliferation marker genes were significantly or extremely significantly decreased (P<0.05 or P<0.01), the cell vitality was also significantly or extremely significantly decreased (P<0.05 or P<0.01), and the newborn cell number was significantly reduced (P<0.05). During the differentiation of ovine preadipocytes, a negative correlation was found between the expressions of miR-128-1-5p and KLF2; After miR-128-1-5p was overexpressed, the mRNA and protein expression of KLF2 were extremely significantly (P<0.01) and significantly (P<0.05) decreased, respectively; The mRNA expressions of differentiation marker genes were significantly or extremely significantly increased (P<0.05 or P<0.01) in the miR-128-1-5p overexpressing cells; More lipid droplets were deposited; The results were opposite after miR-128-1-5p was inhibited. In conclusion, there is a binding site between miR-128-1-5p and KLF2. miR-128-1-5p inhibits the proliferation of ovine preadipocytes. miR-128-1-5p promotes the differentiation of ovine preadipocytes and lipid droplets deposition by targeting and negatively regulating KLF2 in the process of differentiation.

Key words: sheep, preadipocyte, miR-128-1-5p, KLF2, proliferation, differentiation

CLC Number:

S826.2

LIU Jianhua, LIANG Yu, CHE Yutong, WANG Feng, LIU Xuying, ZHAO Bishi, WANG Qiang, AN Lixia, QIAO Liying, LIU Wenzhong. Study on the Regulation of miR-128-1-5p in Proliferation and Differentiation of Ovine Preadipocyte[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1207-1218.

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Study on the Regulation of miR-128-1-5p in Proliferation and Differentiation of Ovine Preadipocyte

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