Division of Geriatrics, Drum Tower Clinic Medical College of Nanjing Medical University, Nanjing, China; Department of Endocrinology and Metabolism, Affiliated Hospital of Nantong University, Nantong, China.
Division of Geriatrics, Drum Tower Clinic Medical College of Nanjing Medical University, Nanjing, China; Department of Endocrinology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, China.
Biochem Biophys Res Commun. 2018 Dec 2;506(4):976-982. doi: 10.1016/j.bbrc.2018.10.197. Epub 2018 Nov 4.
In this study, we investigated whether the GLP-1RA, liraglutide, affected differentiation of C3H10T1/2 mesenchymal stem cells (MSCs) to mature brown adipocytes and involvement of PI3K/AKT/mTOR signaling pathway in this process.
C3H10T1/2 MSCs were induced to differentiate into brown adipocytes and treated with liraglutide (10 nM and 100 nM) for 0, 2, 4, 6 and 8 days with or without PI3K inhibitor LY294002. Oil red O staining was used for lipid droplet staining and cell proliferation was determined by cell counts. Quantitative realtime PCR was employed to determine the expression of adipogenic and mitochondrial genes, mitochondrial DNA (mtDNA). Western blot analyses were used for quantification of protein levels in PI3K/AKT/mTOR signaling pathway.
Liraglutide increased proliferation of C3H10T1/2 MSCs and formation of multilocular lipid droplets during differentiation. Adipogenic and mitochondrial genes, mtDNA were promoted by liraglutide. Moreover, liraglutide treatment increased the levels of phosphorylated AKT and mTOR. LY294002 not only attenuated differentiation of C3H10T1/2 MSCs into brown adipocytes, but also reduced phosphorylated AKT and mTOR levels. However, co-treatment with liraglutide and LY294002 decreased the expression of adipogenic and mitochondrial genes, mtDNA, and phosphorylated AKT and mTOR levels compared to C3H10T1/2 MSCs treated with liraglutide 100 nM.
GLP-1RA promotes brown adipogenesis of C3H10T1/2 mesenchymal stem cells, and PI3K/AKT/mTOR signaling pathway is involved in GLP-1RA-mediated promotion of differentiation.
本研究旨在探讨 GLP-1RA 利拉鲁肽是否影响 C3H10T1/2 间充质干细胞（MSCs）向成熟棕色脂肪细胞分化，并探讨 PI3K/AKT/mTOR 信号通路在此过程中的作用。
将 C3H10T1/2 MSC 诱导分化为棕色脂肪细胞，并分别用 10 nM 和 100 nM 利拉鲁肽处理 0、2、4、6 和 8 天，同时或不用 PI3K 抑制剂 LY294002。油红 O 染色用于检测脂滴染色，细胞计数用于检测细胞增殖。采用实时定量 PCR 检测脂肪生成和线粒体基因、线粒体 DNA（mtDNA）的表达。Western blot 分析用于定量 PI3K/AKT/mTOR 信号通路中的蛋白水平。
利拉鲁肽增加 C3H10T1/2 MSC 的增殖和分化过程中多房脂滴的形成。利拉鲁肽促进脂肪生成和线粒体基因、mtDNA 的表达。此外，利拉鲁肽处理增加了磷酸化 AKT 和 mTOR 的水平。LY294002 不仅减弱了 C3H10T1/2 MSC 向棕色脂肪细胞的分化，还降低了磷酸化 AKT 和 mTOR 的水平。然而，与利拉鲁肽相比，利拉鲁肽和 LY294002 共同处理降低了脂肪生成和线粒体基因、mtDNA 的表达，以及磷酸化 AKT 和 mTOR 的水平。
GLP-1RA 促进 C3H10T1/2 间充质干细胞的棕色脂肪生成，PI3K/AKT/mTOR 信号通路参与 GLP-1RA 介导的分化促进作用。
