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Construction of suppressor of cytokine signaling 2 (SOCS2) adenoviral overexpression vector and its impact on growth-hormone-induced lipolysis in swine primary adipocytes

机译:细胞因子信号传导2(SOCS2)腺病毒过表达载体抑制物的构建及其对生长激素诱导的猪脂肪细胞脂解的影响

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We investigated the effect of overexpression suppressor of cytokine signaling 2 (SOCS2) on lipolysis in swine primary adipocytes (pAd) induced by growth hormone (GH). We constructed pAd-SOCS2 adenoviral overexpression vectors to infect HEK293 cells for virus packaging and propagation. Cultured swine primary adipocytes were infected with virus particles; after 48 h the infected adipocytes were treated with 500 ng GH/mL in the growth medium. Lipometabolism-related gene expressions were detected at 0, 0.25, 0.5, 1, 2, and 4 h, by measuring mRNA and protein levels. The pAd-SOCS2 overexpression vector was successfully constructed and the concentration of titrated virus was 1.2 x 109 PFU/mL. We found that virus infection significantly increased SOCS2 mRNA and protein levels in swine primary adipocytes. Overexpression of SOCS2 significantly inhibited the increase in fatty acid synthase, adipose triglyceride lipase mRNA, and protein expression at 0.5 h. However, after 0.5 h, this inhibition was not significant. We concluded that overexpression of SOCS2 inhibited the increase in lipolysis induced by GH in swine primary adipocytes; this could provide a basis for studies of lipometabolism.
机译:我们调查了细胞因子信号传导2(SOCS2)的过表达抑制因子对生长激素(GH)诱导的猪原代脂肪细胞(pAd)脂解的影响。我们构建了pAd-SOCS2腺病毒过表达载体,以感染HEK293细胞进行病毒包装和繁殖。培养的猪原代脂肪细胞被病毒颗粒感染。 48小时后,在生长培养基中用500ng GH / mL处理感染的脂肪细胞。通过测量mRNA和蛋白质水平,在0、0.25、0.5、1、2和4小时检测到与脂肪代谢相关的基因表达。成功构建了pAd-SOCS2过表达载体,滴定病毒的浓度为1.2 x 109 PFU / mL。我们发现病毒感染显着增加了猪原代脂肪细胞中的SOCS2 mRNA和蛋白水平。 SOCS2的过表达在0.5 h时显着抑制了脂肪酸合酶,脂肪甘油三酸酯脂肪酶mRNA和蛋白质表达的增加。但是,在0.5小时后,这种抑制作用并不明显。我们得出的结论是,SOCS2的过表达抑制了猪原代脂肪细胞中GH诱导的脂解的增加。这可以为脂肪代谢研究提供基础。

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