miR-17-5p调控低氧诱导因子1α介导脂肪细胞分化及血管生成的分子机制  

Molecular mechanism of miR-17-5p regulation of hypoxia inducible factor-1αmediated adipocyte differentiation and angiogenesis

作　　者：刘聪 刘肃 Liu Cong;Liu Su(Department of Plastic Surgery,Affiliated Hospital of Qingdao University,Qingdao 266071,Shandong Province,China)

机构地区：[1]青岛大学附属医院美容整形科,山东省青岛市266071

出　　处：《中国组织工程研究》2021年第7期1069-1074,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:miR-17-5p能够调控脂肪细胞分化,但其作用机制尚未完全明确。低氧诱导因子1α能够促进血管内皮生长因子基因转录促进血管生成,且对脂肪细胞生成具有调控作用,然而低氧诱导因子1α调控脂肪细胞分化和血管生成的具体作用机制尚不清楚。目的:探究miR-17-5p调控低氧诱导因子1α介导脂肪细胞分化及血管生成的分子机制。方法:采用RT-qPCR方法检测miR-17-5p在成熟脂肪细胞中的表达及成熟脂肪细胞中脂肪分化和血管生成相关基因的表达,Western blot检测miR-17-5p抑制剂、pri-miR-17-5p模拟物及低氧诱导因子1α敲降载体转染后脂肪分化和血管生成相关蛋白的表达,油红O染色及MTT检测miR-17-5p抑制剂、pri-miR-17-5p模拟物及低氧诱导因子1α敲降载体转染后脂肪细胞的增殖情况,EGFP报告基因检测miR-17-5p与低氧诱导因子1α的靶向关系。结果与结论:①与未成熟脂肪细胞相比,miR-17-5p在成熟脂肪细胞中高表达(P<0.05),且过表达miR-17-5p促进脂肪细胞增殖(P<0.05),促进脂肪分化和血管生成相关基因的表达(P<0.05);②miR-17-5p与低氧诱导因子1α的3′UTR靶向结合,敲降低氧诱导因子1α能够显著抑制脂肪细胞增殖(P<0.05),下调脂肪分化和血管生成相关基因的表达(P<0.001);③结果表明miR-17-5p通过调控低氧诱导因子1α介导脂肪细胞分化及血管生成。BACKGROUND:miR-17-5p can regulate the differentiation of adipocytes,but its action mechanism is not clear.Hypoxia inducible factor-1αcan promote vascular endothelial growth factor gene transcription and promote angiogenesis,and has a regulatory effect on adipocyte formation.However,the specific mechanism of hypoxia inducible factor-1αin regulating adipocyte differentiation and angiogenesis is not clear.OBJECTIVE:To investigate the molecular mechanism of miR-17-5p regulation of hypoxia inducible factor-1αmediated adipocyte differentiation and angiogenesis.METHODS:miR-17-5p expression level and adipocyte differentiation and the expression of angiogenesis markers in mature adipocytes were verified by RTqPCR.The adipocyte differentiation and angiogenesis markers expression of adipocyte transfecting with miR-17-5p inhibitor,pri-miR-17-5p mimic and hypoxia inducible factor-1αknockdown vector were determined by western blot assay.The proliferation of adipocytes was detected by Oil Red O staining assay and MTT assay after transfection with miR-17-5p inhibitor,pri-miR-17-5p mimic and hypoxia inducible factor-1αknockdown vector.The predicted relationship between miR-17-5p and hypoxia inducible factor-1αwas further verified by EGFP report gene assay.RESULTS AND CONCLUSION:(1)Compared to immature adipocyte,miR-17-5p was highly expressed in mature adipocyte(P<0.05);overexpression of miR-17-5p increased the adipocyte proliferation(P<0.05),and upregulated adipocyte differentiation and angiogenesis markers level(P<0.05).(2)The miR-17-5p directly targeted with hypoxia inducible factor-1α3′UTR;knockdown hypoxia inducible factor-1αinhibited the adipocyte proliferation(P<0.05),and decreased adipocyte differentiation and angiogenesis markers level(P<0.001).(3)It is concluded that miR-17-5p regulation of hypoxia inducible factor-1αmediated adipocyte differentiation and angiogenesis.

关 键 词：MIRNA miR-17-5p 因子 低氧诱导因子1Α 脂肪细胞 血管生成 细胞增殖 靶向 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]