miR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the <i>PDE4B</i> gene.

Liyi Zhang; Xin LI; Qing XU; Xinzhu Huang; Yanyan LI; Wei Liu; Youli Wang; Yaqiu Lin

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miR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.

Author: Liyi ZHANG ¹ ; Xin LI ¹ ; Qing XU ¹ ; Xinzhu HUANG ¹ ; Yanyan LI ¹ ; Wei LIU ¹ ; Youli WANG ¹ ; Yaqiu LIN ¹
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1. College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, Sichuan, China.
Publication Type:Journal Article
Keywords: PDE4B; dual luciferase reporter assay; goat; intramuscular adipocyte differentiation; miR-23b-3p
MeSH: Animals; MicroRNAs/metabolism*; Goats/genetics*; PPAR gamma/metabolism*; Adipogenesis/genetics*; Cell Differentiation/genetics*; Luciferases; RNA, Messenger
From: Chinese Journal of Biotechnology 2023;39(12):4887-4900
CountryChina
Language:Chinese
Abstract: This study aimed to explore the effect of miR-23b-3p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-23b-3p plays its roles via targeting the PDE4B gene. Based on the pre-transcriptome sequencing data obtained previously, the miR-23b-3p, which was differentially expressed in goat intramuscular adipocytes before and after differentiation, was used as an entry point. real-time quantitative-polymerase chain reaction (qPCR) was used to detect the expression pattern of miR-23b-3p during the differentiation of goat intramuscular preadipocytes. The effects of miR-23b-3p on adipose differentiation and adipose differentiation marker genes were determined at the morphological and molecular levels. The downstream target genes of miR-23b-3p were determined using bioinformatics prediction as well as dual luciferase reporter assay to clarify the targeting relationship between miR-23b-3p and the predicted target genes. The results indicated that overexpression of miR-23b-3p reduced lipid droplet accumulation in goat intramuscular adipocytes, significantly down-regulated the expression levels of adipogenic marker genes AP2, C/EBPα, FASN, and LPL (P < 0.01). In addition, the expressions of C/EBPβ, DGAT2, GLUT4 and PPARγ were significantly downregulated (P < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of ACC, ATGL, AP2, DGAT2, GLUT4, FASN and SREBP1 were extremely significantly up-regulated (P < 0.01), and the expression levels of C/EBPβ, LPL and PPARγ were significantly up-regulated (P < 0.05). It was predicted that PDE4B might be a target gene of miR-23b-3p. The mRNA expression level of PDE4B was significantly decreased after overexpression of miR-23b-3p (P < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of PDE4B (P < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with PDE4B gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the PDE4B gene.