摘要
目的调查小鼠原代肝窦内皮细胞(liver sinusoidal endothelial cells,LSECs)中NOD2信号通路激活后产生的免疫应答效应。方法使用胶原酶灌注方法结合percoll密度梯度离心加抗小鼠LSEC免疫磁珠分离纯化小鼠LSEC。定量PCR检测小鼠原代LSEC NOD1和NOD2 mRNA表达水平,同时给胞壁酰二肽刺激小鼠原代LSEC;定量PCR检测NOD2、RIP2、TNF-α和IL-6mRNA表达水平;定量ELISA检测细胞上清IL-6和TNF-α蛋白产量。结果小鼠原代LSEC中NOD2基础表达水平相对于NOD1基础表达水平较低。胞壁酰二肽刺激LSEC后诱导NOD2及其下游分子RIP2表达水平上调,接着诱导产生IL-6效应分子,而没有诱导产生TNF-α等其它细胞因子和趋化因子。结论胞壁酰二肽能够激活小鼠LSEC中NOD2介导的信号通路,并诱导产生IL-6效应分子,可能在维持肝脏内环境稳定中发挥着重要作用。
This study was designed to investigate the effects of the immune response mediated by NOD2 receptor in liver sinusoidal endothelial cells（LSEC） of mice. Firstly, we used the methods of the collagenase perfusion, percoll density gradient centrifugation and anti-LSEC MicroBeads separation and purification to obtain high purity LSEC. Then the LSECs were stimulated with the muramyl-dipeptide（MDP） or not. Total RNA was isolated from LSECs for detecting the mRNA of NOD1, NOD2, RIP2, TNF-α and IL-6 by qRT-PCR. The cytokines（IL-6 and TNF-α） in the cells supernatant were detected by quantitative ELISA. Data showed that NOD2 mRNA expression was relative lower at baseline in primary LSECs as compared with NOD1, and stimulation of LSECs with MDP could induce up-regulation of NOD2 and RIP2 mRNA expression and the production of IL-6 proteins, but not other cytokines or chemokines, such as TNF-α. In conclusion, MDP activates the NOD2 signaling pathway in LSEC and induces the production of the effector molecule IL-6, thus plays a key role in maintaining liver homeostasis.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第11期1025-1028,共4页
Immunological Journal
基金
国家自然科学基金(81001313)
江西省卫生厅科技计划(20143077)
关键词
NOD2
肝窦内皮细胞
胞壁酰二肽
信号通路
NOD2
Liver sinusoidal endothelial cell
Muramyl-dipeptide
Signaling pathway
