Molecular mechanism of mir-17-5p regulation of hypoxia inducible factor-1α mediated adipocyte differentiation and angiogenesis / 中国组织工程研究

ABSTRACT

BACKGROUND: miR-17-5p can regulate the differentiation of adipocytes, but its action mechanism is not clear. Hypoxia inducible factor-1α can 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-1α in regulating adipocyte differentiation and angiogenesis is not clear. OBJECTIVE: To investigate the molecular mechanism of miR-17-5p regulation of hypoxia inducible factor-1α mediated 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 RT-qPCR. The adipocyte differentiation and angiogenesis markers expression of adipocyte transfecting with miR-17-5p inhibitor, pri-miR-17-5p mimic and hypoxia inducible factor-1α knockdown 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-1α knockdown vector. The predicted relationship between miR-17-5p and hypoxia inducible factor-1α was 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-1α 3′UTR; knockdown hypoxia inducible factor-1α inhibited 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-1α mediated adipocyte differentiation and angiogenesis.