p38MAPK在重症急性胰腺炎大鼠枯否细胞分泌促炎细胞因子中的作用

Role of p38MAPK in production of pro-inflammatory cytokines in Kupffer cells from severe acute pancreatitis rats

目的 :探讨p38MAPK信号转导通路在重症急性胰腺炎 (SAP)大鼠枯否细胞 (KCs)分泌促炎细胞因子TNF -α和IL - 1β中的作用。 方法 :30只SD大鼠随机分为 :①假手术对照 (SO)组 ;②SAP组 ;③SAP +CNI - 14 93(p38MAPK抑制剂 )组。SAP模型通过胰胆管逆行注射 5 %牛磺胆酸钠诱导。假手术或造模后 12h处死动物分离出KCs,采用实时定量PCR方法检测KCs内TNF -α和IL - 1βmRNA的表达 ,采用Westernblot法检测p38MAPK活化情况 ,并用ELISA法检测血浆的TNF -α和IL - 1β含量。结果 :SAP大鼠KCs内TNF -α和IL - 1βmRNA的表达明显强于假手术组 ,p38MAPK活性显著高于SO组 ,同时血浆TNF -α和IL - 1β含量明显高于SO组 ,使用CNI - 14 93的SAP大鼠上述指标均显著低于SAP组。结论 :p38MAPK信号转导通路介导了SAP大鼠的KCs促炎细胞因子TNF -α和IL- 1β的分泌 。

AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the Kupffer cells (KCs) production of pro-inflammatory cytokines, tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β), in severe acute pancreatitis (SAP) rats. METHODS: Sprague-Dwaley rats were randomized into three groups: ①sham operation rats, ②SAP rats, ③SAP rats given the p38 MAPK inhibitor CNI-1493(10 mg/kg, iv). The SAP model was induced by the bili-pancreatic duct infusion with 5% sterile soduim taurocholate solution. Rats from each group were killed at 12 h after sham operation or SAP and Kupffer cells (KCs) were isolated. The mRNA expressions of TNF-α and IL-1β (by quantitative real-time RT-PCR) and p38 MAPK activity (by Western blot analysis) in KCs were examined. The levels of TNF-α and IL-1β in plasma were determined by ELISA. RESULTS: There was a significant acvitation of p38 MAPK in KCs harvested from SAP rats than those from sham operation rats. SAP also promoted the mRNA expressions of TNF-α and IL-1β in KCs and the plasma levels of TNF-α and IL-1β. These events were significantly inhibited by treatment with CNI-1493.CONCLUSIONS: p38 MAPK activation is one important aspect of the signaling events that may mediate the KCs production of pro-inflammatory cytokines, TNF-α and IL-1β, in SAP rats. The inhibition of the p38 MAPK may be a potential target in the prevention and treatment of SAP.