【摘要】 目的用胰腺炎相关性腹水(PAAF)刺激人单核细胞,探讨PPARγ激动剂吡格列酮对细胞炎症信号通路的影响。方法 PAAF收集自重症胰腺炎患者。将体外培养的人单核细胞THP-1分为4组:对照组(C组)、实验组(A组)、吡格列酮组(P组)、GW9662(G组)。A组用PAAF刺激细胞;P组加入终浓度为20μmol/L吡格列酮,2h后再用PAAF刺激细胞;G组先加入PPARγ拮抗剂GW9662,30min后加入吡格列酮,2h后再用PAAF刺激细胞。12h后收集4组细胞,采用实时定量PCR方法检测细胞TNF-α、IL-6的mRNA表达,蛋白质印迹法检测NF-κB p65、p38MAPK的活化程度和IκB蛋白表达变化。结果 PAAF刺激后,A组细胞促炎细胞因子TNF-α、IL-6的mRNA表达增加,NF-κB、p38MAPK活化,IκB蛋白水平降低,与C组相比差异有统计学意义(P<0.05);P组经吡格列酮干预后,TNF-α、IL-6的mRNA表达降低,NF-κB、p38MAPK活性下调,IκB蛋白表达升高,与A组比较差异有统计学意义(P<0.05);G组应用GW9662后可拮抗吡格列酮降低促炎细胞因子的作用,TNF-α、IL-6的mRNA表达增加,NF-κB、p38MAPK活化,IκB蛋白水平降低,与P组相比差异有统计学意义(P<0.05)。结论吡格列酮可能通过抑制NF-κB、p38MAPK炎症信号通路活化,减少促炎细胞因子产生,控制炎症发生发展。更多还原

【Abstract】 Objective To investigate the inhibitory effect of pioglitazone on inflammatory signaling pathways induced by human pancreatitis-associated ascitic fluid(PAAF) in human monocytic cell line THP-1.Methods PAAF was collected from patients with severe acute pancreatitis.The cultured THP-1 cells were divided into 4 groups: control group(C),PAAF group(A),PPARγ agonist pioglitazone group(P),and PPARγ antagonist GW9662 group(G).Cells in group A were stimulated with PAAF,those in group P were treated with 20 μmol/L PPARγ agonist pioglitazone 2 h after stimulation with PAAF,and those in group G were treated with PPARγ antagonist GW9662,pioglitazone(30 min later) and PAAF(2 h later) in turn.After incubation for 12 h,TNF-α and IL-6 mRNA expression was measured by real-time PCR,and Western blotting analysis was used to examine the expression of the phospho-p38-mitogen-activated protein kinase(MAPK),nuclear transcription factor(NF-κB) p65 levels,and IκB. Results Compared with group C,TNF-α and IL-6 mRNA expression in group A was increased,NF-κB and p38MAPK protein levels were increased,and IκB protein level was decreased(P<0.05).Compared with group A,TNF-α and IL-6 mRNA expression was decreased in group P,NF-κB and p38MAPK protein levels were down-regulated,and IκB protein level was increased(P<0.05).Compared with group P,TNF-α and IL-6 mRNA expression was increased in group G,NF-κB and p38MAPK protein levels were increased,and IκB protein level was decreased(P<0.05).Conclusion The results show that pioglitazone can inhibit PAAF-stimulated THP-1 inflammation by blocking p38MAPK and NF-κB signaling pathway.更多还原