| 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 |
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| 引用本文： | 刘建华,梁煜,车雨彤,王凤,刘旭莹,赵弼时,王强,安丽霞,乔利英,刘文忠.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[J].畜牧兽医学报,2020,51(6):1207-1218. |
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| 作者姓名： | 刘建华 梁煜 车雨彤 王凤 刘旭莹 赵弼时 王强 安丽霞 乔利英 刘文忠 |
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| 作者单位： | 山西农业大学动物科技学院, 太谷 030801 |
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| 基金项目： | 国家自然科学基金（31972560）；山西省“1331工程”重点学科建设计划（J201811301） |
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| 摘 要： | 旨在研究miR-128-1-5p对绵羊前体脂肪细胞增殖与分化的调节作用。本研究经理论预测和试验研究验证miR-128-1-5p的靶基因;过表达miR-128-1-5p后,用qPCR检测增殖标志基因的表达,CCK-8和EdU检测细胞增殖情况;用qPCR和Western blotting检测miR-128-1-5p和其靶基因在前体脂肪细胞分化中的表达趋势;通过过表达或抑制miR-128-1-5p研究其对绵羊前体脂肪细胞分化的调节机制;用油红O染色检测成脂能力。结果表明,KLF2是miR-128-1-5p的靶基因。前体脂肪细胞增殖过程中,过表达miR-128-1-5p后,增殖标志基因的表达量显著或极显著降低(P<0.05或P<0.01),细胞活力显著或极显著降低(P<0.05或P<0.01),新生细胞数显著减少(P<0.05)。前体脂肪细胞分化过程中,miR-128-1-5p与KLF2的表达呈负相关;过表达miR-128-1-5p极显著下调了KLF2 mRNA的表达量(P<0.01),显著下调了KLF2蛋白的表达量(P<0.05),显著或极显著上调了分化标志基因mRNA的表达(P<0.05或P<0.01),产生更多脂滴;抑制miR-128-1-5p则结果相反。综上所述,miR-128-1-5p与KLF2存在结合位点。miR-128-1-5p抑制绵羊前体脂肪细胞的增殖,在分化过程中通过靶向抑制KLF2的表达,促进前体脂肪细胞分化和脂滴沉积。
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| 关 键 词： | 绵羊 前体脂肪细胞 miR-128-1-5p KLF2 增殖 分化 |
| 收稿时间： | 2019-12-04 |
Study on the Regulation of miR-128-1-5p in Proliferation and Differentiation of Ovine Preadipocyte |
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| 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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| Authors: | LIU Jianhua LIANG Yu CHE Yutong WANG Feng LIU Xuying ZHAO Bishi WANG Qiang AN Lixia QIAO Liying LIU Wenzhong |
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| Institution: | College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China |
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| 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. |
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| Keywords: | sheep preadipocyte miR-128-1-5p KLF2 proliferation differentiation |
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