Zhang Liyi, Li Xin, Xu Qing, Huang Xinzhu, Li Yanyan, Liu Wei, Wang Youli, Lin Yaqiu
College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, Sichuan, China.
Sichuan Province Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, Sichuan, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Dec 25;39(12):4887-4900. doi: 10.13345/j.cjb.230156.
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 targeting the 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 2, /, , and ( < 0.01). In addition, the expressions of /, 2, 4 and were significantly downregulated ( < 0.05). After interfering with the expression of miR-23b-3p, lipid droplet accumulation was increased in goat intramuscular adipocytes. The expression levels of , , 2, 2, 4, and 1 were extremely significantly up-regulated ( < 0.01), and the expression levels of /, and were significantly up-regulated ( < 0.05). It was predicted that might be a target gene of miR-23b-3p. The mRNA expression level of was significantly decreased after overexpression of miR-23b-3p ( < 0.01), and the interference with miR-23b-3p significantly increased the mRNA level of ( < 0.05). The dual luciferase reporter assay indicated that miR-23b-3p had a targeting relationship with gene. MiR-23b-3p regulates the differentiation of goat intramuscular preadipocytes by targeting the gene.
本研究旨在探讨miR-23b-3p对山羊肌内前体脂肪细胞分化的影响，并证实miR-23b-3p是否通过靶向该基因发挥作用。以前期获得的转录组测序数据为基础，选取在山羊肌内脂肪细胞分化前后差异表达的miR-23b-3p作为切入点，采用实时定量聚合酶链反应（qPCR）检测miR-23b-3p在山羊肌内前体脂肪细胞分化过程中的表达模式，从形态学和分子水平确定miR-23b-3p对脂肪分化及脂肪分化标志基因的影响。利用生物信息学预测及双荧光素酶报告基因检测确定miR-23b-3p的下游靶基因，以阐明miR-23b-3p与预测靶基因之间的靶向关系。结果表明，miR-23b-3p过表达减少了山羊肌内脂肪细胞中脂滴的积累，显著下调了成脂标志基因2、/、、和的表达水平（<0.01），此外，/、2、4和的表达也显著下调（<0.05）。干扰miR-23b-3p的表达后，山羊肌内脂肪细胞中脂滴积累增加，、、2、2、4、和1的表达水平极显著上调（<0.01），/、和的表达水平显著上调（<0.05）。预测可能是miR-23b-3p的靶基因，miR-23b-3p过表达后，的mRNA表达水平显著降低（<0.01），干扰miR-23b-3p后，的mRNA水平显著升高（<0.05）。双荧光素酶报告基因检测表明，miR-23b-3p与基因存在靶向关系。miR-23b-3p通过靶向基因调控山羊肌内前体脂肪细胞的分化。
