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.

中文翻译：

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lmerRNA Meg3通过海绵化miR-217调节Dickkopf-3来介导3种T3-L1前脂肪细胞中凯莫瑞诱导的血管生成和脂肪生成。

目的肥胖症通常是由过多的脂肪生成引起的，并受长的非编码RNA（lncRNA）和microRNA（miRNA）的调节。我们进行了这项研究，以调查Meg3表达对脂肪形成的影响，以及Meg3 / miR-217 / Dkk3轴介导的脂肪形成和血管生成的分子机制。方法将3种T3-L1前脂肪细胞与凯莫瑞一起孵育，并用过表达Meg3（OE-Meg3）和过表达Dkk3（OE-Dkk3）的质粒，siRNA和miR-217模拟物，抑制剂和加扰序列转染48 h或72 h 。用相应的方法检测3种T3-L1前脂肪细胞的细胞增殖，脂肪形成和血管生成能力的变化。通过蛋白质印迹法检测相关蛋白的表达。结果Chemerin降低了miR-217的表达并增加了Meg3的表达，同时促进了3种T3-L1前脂肪细胞的增殖，脂肪生成和血管生成。此外，OE-Meg3与凯莫瑞共同处理时，对3个T3-L1前脂肪细胞具有协同作用。使用双重荧光素酶报告系统验证了Meg3与miR-217之间以及miR-217与Dkk3之间的靶相互作用。SiMeg3拮抗凯莫瑞诱导的变化，而miR-217抑制剂的加入则减弱了3种T3-L1前脂肪细胞中siMeg3诱导的变化。miR-217模拟物抑制了3个T3-L1前脂肪细胞的增殖，脂肪形成和血管生成，而OE-Dkk3 Chemerin促进了T3-L1前脂肪细胞的增殖，脂肪形成和血管生成，从而促进了脂肪酸结合蛋白4和血管内皮生长因子（VEGF）蛋白的表达，并降低了cyclin D1，c-Myc和β-catenin蛋白的表达 同时，OE-Meg3或OE-Dkk3进一步增强了这些作用。但是，siMeg3或miR-217模拟物或siDkk3的转染逆转了先前的变化。结论Meg3 / miR-217 / Dkk3通过激活VEGF信号通路和抑制Wnt /β-catenin信号通路诱导3个T3-L1前脂肪细胞的脂肪生成和血管生成。