基础医学与临床 ›› 2018, Vol. 38 ›› Issue (10): 1412-1416.

• 研究论文 • 上一篇    下一篇

杨梅素联合围脂滴蛋白基因沉默促进前脂肪细胞系的降脂

张少华,王君实,董维鹏,燕炯   

  1. 山西医科大学
  • 收稿日期:2017-10-09 修回日期:2018-01-25 出版日期:2018-10-05 发布日期:2018-09-28
  • 通讯作者: 燕炯 E-mail:yanjiong@126.com
  • 基金资助:
    山西医科大学科技创新基金项目

Myricetin combined with PLIN1 silencing promotes lipolysis of 3T3-L1 adipocytes

  • Received:2017-10-09 Revised:2018-01-25 Online:2018-10-05 Published:2018-09-28

摘要: 目的 探讨杨梅素(Myric)与围脂滴蛋白(PLIN1)基因沉默对3T3-L1脂肪细胞脂解通路的影响。方法 采用经典“鸡尾酒”方法诱导分化3T3-L1前脂肪细胞,筛选出Myric最佳干预浓度和时间。Myric联合sh-PLIN1干扰载体对成熟的脂肪细胞进行干预,实验共分为4组:对照组(control group)、转染组(sh-RNA group)、杨梅素组(Myric group)和联合干预组(Myric+sh-RNA group)。油红O染色脂滴,观察脂滴形态;Western blot检测细胞中ERK、p-ERK、MEK、p-MEK和激素敏感性脂肪酶磷酸化蛋白(p-HSL)的表达量;ELISA测定细胞中环磷酸腺苷(cAMP)、蛋白激酶A(PKA)和磷酸化的蛋白激酶C(p-PKC)的含量。结果 干预后,与Myric组和sh-RNA组相比,Myric+sh-RNA组脂滴形态变小,数量明显减少。与sh-RNA组和对照组相比,Myric+sh-RNA组和Myric组细胞内p-PKC含量,p-ERK/ERK和p-MEK/MEK的比值以及p-HSL表达量显著升高(P<0.05)。同时,Myric+sh-RNA组和Myric组细胞内cAMP和PKA含量低于sh-RNA组和对照组(P<0.05)。结论 Myric的促脂解作用可能是通过激活PKC-MEK/ERK信号通路,增加该通路中p-HSL的表达量来实现的;Myric同时可能对cAMP/PKA信号通路中相关因子的活性有一定的抑制作用。

关键词: 杨梅素, PLIN1基因沉默, 3T3-L1脂肪细胞, 脂解机制

Abstract: Objective To investigate the mechanism of myricetin combined with PLIN1 silencing on lipolysis of 3T3-L1 adipocytes. Methods 3T3-L1 adipocytes were routinely induced to differentiate by the hormonal cocktail. According to the optimal concentration of Myric combined with the high efficient transfection vector of sh-PLIN1, the intervention experiment was divided into four groups: control group, transfection group (sh-RNA), Myricetin group (Myric) and combination group (Myric+sh-RNA). The Oil Red O staining was used to observe the morphology of lipid droplets. The protein expression levels of ERK, p-ERK, MEK, p-MEK and p-HSL were detected by Western blot. Enzyme linked immunosorbent assay (ELISA) was used to measure the content of the cyclic adenosine monophosphate (cAMP), protein kinase A (PKA) and p-PKC. Results Compared to Myric group and sh-RNA group, size of lipid droplets decreased obviously in Myric+sh-RNA group. Compared to sh-RNA group and control group,the relative value of p-PKC, p-ERK/ERK, p-MEK/MEK and p-HSL increased significantly in Myric+sh-RNA group and Myric group (P<0.05), meanwhile, the content of cAMP and PKA decreased significantly (P<0.05). Conclusions Myric may promote lipolysis through the activation of PKC-MEK/ERK signaling pathway, and further increase expression level of p-HSL in this pathway. Meanwhile, Myric may also inhibit the activity of related factors in cAMP/PKA signaling pathway.

Key words: Myricetin, Perilipin 1 gene silencing, 3T3-L1 adipocytes, signal pathways of lipolysis

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