摘要
目的:研究PYNOD对脂多糖(lipopolysaccharide,LPS)介导的BV2小胶质细胞极化的影响,并探讨其作用机制。方法:体外培养BV2细胞,建立LPS介导的细胞模型,并将细胞分成对照(control)组、LPS组、空质粒对照组(NC+LPS组)和过表达PYNOD组(PYNOD+LPS组)。采用real-time PCR检测白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、IL-10和PYNOD的mRNA表达;ELISA检测细胞培养液中IL-6、TNF-α和IL-10含量;Western blot检测细胞中PYNOD、核因子κB(nuclear factor-κB,NF-κB)P65、磷酸化P65(p-P65)和过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)的蛋白表达。结果:与con⁃trol组相比,LPS组IL-6、TNF-α和IL-10的表达和分泌量均显著升高(P<0.05);p-P65蛋白水平显著升高,而PPARγ蛋白表达显著降低(P<0.01)。与NC+LPS组相比,PYNOD+LPS组IL-6和TNF-α的表达和分泌量显著降低(P<0.05),IL-10的表达和分泌量显著升高(P<0.05);p-P65蛋白水平显著降低,而PPARγ蛋白表达显著升高(P<0.05)。结论:PYNOD过表达能够抑制小胶质细胞的M1型极化,促进M2型极化,其机制可能与降低NF-κB活性和升高PPARγ表达有关。
AIM:To investigate the effect of PYNOD on BV2 microglia polarization induced by lipopolysac⁃charide(LPS),and to explore the related mechanism.METHODS:The BV2 cells were randomly divided into control group,LPS group,negative control(NC)+LPS group and PYNOD overexpression(PYNOD+LPS)group.The mRNA levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),IL-10 and PYNOD were measured by real time-PCR.The concentrations of IL-6,TNF-αand IL-10 in cell culture supernatants were measured by ELISA.Western blot was used to detect the protein levels of PYNOD,nuclear factor-κB(NF-κB)P65,phosphorylated P65(p-P65)and peroxi⁃some proliferator-activated receptorγ(PPARγ).RESULTS:Compared with control group,LPS increased the expres⁃sion and release levels of IL-6,TNF-αand IL-10(P<0.05),and stimulated the phosphorylation of P65,but decreased PPARγprotein expression(P<0.05).Compared with NC+LPS group,the expression and release levels of IL-6 and TNF-αin PYNOD+LPS group were decreased(P<0.05),while the expression and release levels of IL-10 were increased(P<0.05).The protein level of p-P65 was decreased,while the protein expression of PPARγwas increased(P<0.05).CON⁃CLUSION:Overexpressed PYNOD inhibits M1 polarization and promotes M2 polarization of microglia,and the corresponding mechanism is involved in the inhibition of NF-κB and stimulation of PPARγ.
作者
曾琪
俞梅琴
刘艳娟
ZENG Qi;YU Mei-qin;LIU Yan-juan(Department of Ultrasound,the First Affiliated Hospital of Gannan Medical College,Ganzhou 341001,China;Gannan Medical College,Ganzhou 341001,China;Institute of Emergency Medicine,Hunan Provincial Key Laboratory of Emer-gency and Critical Care Metabonomics,Hunan Provincial People's Hospital/the First Affiliated Hospital of Hunan Normal University,Changsha 410005,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2022年第4期645-650,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81960298
No.82102277
No.82102278)
湖南省卫健委科研项目(No.202210002585)。
作者简介
通讯作者:刘艳娟,Tel:0731-83929129,E-mail:877497821@qq.com。
