GITRL在内毒素诱导的Kupffer细胞凋亡中的作用研究

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。

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

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.