GITRL在内毒素诱导的Kupffer细胞凋亡中的作用研究

Role of glucocorticoid-induced tumor necrosis factor-related protein ligand(GITRL) on lipopolysaccharide induced Kupffer cells apoptosis

目的:探讨糖皮质激素诱导的肿瘤坏死因子相关蛋白配体(GITRL)在脂多糖(LPS)诱导的Kupffer细胞(KCs)凋亡中的作用。方法:分离BALB/c小鼠的KCs,转染对照siR-NA或者GITRL siRNA 24 h后,分四组培养,分别为对照(Control)组:仅加入培养液;地塞米松(Dex)组:加入Dex10μmol/L;LPS组:加入LPS 1 mg/L;LPS+Dex组:加入LPS 1 mg/L和Dex 10μmol/L。24 h后用免疫细胞化学法检测GITRL蛋白的表达,应用Annexin V/PI双染标记和流式细胞术检测KCs的凋亡率。结果:LPS刺激增加了KCs GITRL的表达(P<0.05),然而地塞米松处理降低了LPS诱导的GITRL表达。LPS刺激诱导了KCs的凋亡,但是沉默GITRL基因或者地塞米松处理抑制了LPS诱导的凋亡(P<0.05)。结论:LPS可以诱导小鼠KCs的凋亡,其作用可能依赖于GITRL信号的转导。

AIM： To study the role of glucocorticoid-induced tumor necrosis factor-related protein ligand（GITRL） on apoptosis of mouse Kupffer cells（KCs） induced by lipopolysaccharide（LPS）.METHODS： The KCs were isolated from BALB/c mice and transfected with Control siRNA or GITRL siRNA for 24 h.The KCs were randomly divided into four groups including control group： cultured in media alone,dexamethasone（Dex） group： media with Dex 10 μmol/L,LPS group： media with LPS 1 mg/L,and LPS＋Dex group： media with LPS 1 mg/L and Dex 10 μmol/L.At 24 h after treatment,the expression of GITRL was detected by immunocytochemistry.The apoptosis of KCs was measured by Annexin V-FITC/PI double staining and FCM.RESULTS： The GITRL expression of KCs was increased by LPS challenge（P〈0.05）,whereas Dex treatment attenuated the increase.LPS challenge induced KCs apoptosis,but the LPS induced apoptosis was inhibited by GITRL siRNA transfection or Dex treatment（P〈0.05,respectively）.CONCLUSION： LPS could induce mouse KCs apoptosis,which may be depend on GITRL signal transduction.