Xianwei Huang1, Caihua Fu2, Wenhui Liu3, Yansheng Liang3, Peilun Li4, Zhiquan Liu5, Qiping Sheng3, Ping Liu6. 1. Department of Cardiology, The Second Hospital of Shandong University, Jinan, Shandong 250033, China; Department of Emergency, the First Affiliated Hospital of Xiamen University, Xiamen, Fujian 361003, China. 2. Department of Cardiology, Jinan central hospital affiliated Shandong University, Jinan, Shandong 250012, China. 3. Department of Cardiology, The Second Hospital of Shandong University, Jinan, Shandong 250033, China. 4. Department of Cardiology, Linyi People's Hospital, Linyi, Shandong 276003, China. 5. Department of Cardiology, The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital), Hefei, Anhui 230001,China. 6. Department of Cardiology, The Second Hospital of Shandong University, Jinan, Shandong 250033, China. Electronic address: lping@sdu.edu.cn.
Abstract
PURPOSE: Obesity is often caused by the excess adipogenesis and regulated by long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We performed this study to investigate the influence of Meg3 expression on adipogenesis and also the Meg3/miR-217/Dkk3 axis-mediated molecular mechanism in adipogenesis and angiogenesis. METHODS: 3 T3-L1 preadipocytes were incubated with chemerin and transfected with Meg3-overexpressing (OE-Meg3) and Dkk3-overexpressing (OE-Dkk3) plasmids, siRNAs, and miR-217 mimics, inhibitor and scrambled sequences for 48 h or 72 h. The changes in cell proliferation, adipogenesis and angiogenesis ability in 3 T3-L1 preadipocytes was detected by using the corresponding assay. The expressions of related proteins were detected via western blot. RESULTS: Chemerin decreased miR-217 expression and increased Meg3 expression, meanwhile promoted the proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes. Besides, OE-Meg3 exerted the synergistic effect on 3 T3-L1 preadipocytes when co-treated with chemerin. The target interactions between Meg3 and miR-217 as well as between miR-217 and Dkk3 were validated using dual-luciferase reporter system. SiMeg3 antagonized chemerin-induced changes, while the addition of miR-217 inhibitor attenuated siMeg3-induced changes in 3 T3-L1 preadipocytes. The proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes were suppressed by miR-217 mimics, while promoted by the OE-Dkk3 Chemerin promoted the expression of fatty acid binding protein 4 and vascular endothelial growth factor (VEGF) proteins, and decreased the expression of cyclin D1, c-Myc, and β-catenin proteins. Meanwhile, these effects were further enhanced by OE-Meg3 or OE-Dkk3. However, the transfection of siMeg3, or miR-217 mimics, or siDkk3 reversed the previous changes. CONCLUSIONS: Meg3/miR-217/Dkk3 induced adipogenesis and angiogenesis in 3 T3-L1 preadipocytes via activating VEGF signaling pathway and inhibiting Wnt/β-catenin signaling pathway.
PURPOSE: Obesity is often caused by the excess adipogenesis and regulated by long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). We performed this study to investigate the influence of Meg3 expression on adipogenesis and also the Meg3/miR-217/Dkk3 axis-mediated molecular mechanism in adipogenesis and angiogenesis. METHODS: 3 T3-L1 preadipocytes were incubated with chemerin and transfected with Meg3-overexpressing (OE-Meg3) and Dkk3-overexpressing (OE-Dkk3) plasmids, siRNAs, and miR-217 mimics, inhibitor and scrambled sequences for 48 h or 72 h. The changes in cell proliferation, adipogenesis and angiogenesis ability in 3 T3-L1 preadipocytes was detected by using the corresponding assay. The expressions of related proteins were detected via western blot. RESULTS:Chemerin decreased miR-217 expression and increased Meg3 expression, meanwhile promoted the proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes. Besides, OE-Meg3 exerted the synergistic effect on 3 T3-L1 preadipocytes when co-treated with chemerin. The target interactions between Meg3 and miR-217 as well as between miR-217 and Dkk3 were validated using dual-luciferase reporter system. SiMeg3 antagonized chemerin-induced changes, while the addition of miR-217 inhibitor attenuated siMeg3-induced changes in 3 T3-L1 preadipocytes. The proliferation, adipogenesis and angiogenesis in 3 T3-L1 preadipocytes were suppressed by miR-217 mimics, while promoted by the OE-Dkk3Chemerin promoted the expression of fatty acid binding protein 4 and vascular endothelial growth factor (VEGF) proteins, and decreased the expression of cyclin D1, c-Myc, and β-catenin proteins. Meanwhile, these effects were further enhanced by OE-Meg3 or OE-Dkk3. However, the transfection of siMeg3, or miR-217 mimics, or siDkk3 reversed the previous changes. CONCLUSIONS:Meg3/miR-217/Dkk3 induced adipogenesis and angiogenesis in 3 T3-L1 preadipocytes via activating VEGF signaling pathway and inhibiting Wnt/β-catenin signaling pathway.
Authors: Roderick C Slieker; Louise A Donnelly; Hugo Fitipaldi; Gerard A Bouland; Giuseppe N Giordano; Mikael Åkerlund; Mathias J Gerl; Emma Ahlqvist; Ashfaq Ali; Iulian Dragan; Petra Elders; Andreas Festa; Michael K Hansen; Amber A van der Heijden; Dina Mansour Aly; Min Kim; Dmitry Kuznetsov; Florence Mehl; Christian Klose; Kai Simons; Imre Pavo; Timothy J Pullen; Tommi Suvitaival; Asger Wretlind; Peter Rossing; Valeriya Lyssenko; Cristina Legido Quigley; Leif Groop; Bernard Thorens; Paul W Franks; Mark Ibberson; Guy A Rutter; Joline W J Beulens; Leen M 't Hart; Ewan R Pearson Journal: Diabetes Date: 2021-08-10 Impact factor: 9.461