| 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20 μmol/L吡格列酮,2 h后再用PAAF刺激细胞；G组先加入PPARγ拮抗剂GW9662,30 min后加入吡格列酮,2 h后再用PAAF刺激细胞。12 h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。 |
| 关键词: 吡格列酮 胰腺炎 单核细胞 NF-κB p38丝裂原活化蛋白激酶类 |
| DOI：10.3724/SP.J.1008.2012.001182 |
| 投稿时间：2012-06-13修订日期：2012-10-31 |
| 基金项目:上海市自然科学基金(09ZR1429100),上海市松江区卫生局“医学领先合作项目”. |
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| Effect of pioglitazone on inflammatory signaling pathways induced by pancreatitis-associated ascitic fluid in human monocytic cell line THP-1 |
| XU Ping1*,WANG Jing1,HOU Yan-qiang2,LI Qing-hua1,CHEN Cheng3,LOU Xiao-li2 |
(1. Department of Gastroenterology, Songjiang Branch, The First People’s Hospital of Shanghai, Shanghai Jiaotong University, Shanghai 201600, China
2. Central Laboratory, Songjiang Branch, The First People’s Hospital of Shanghai, Shanghai Jiaotong University, Shanghai 201600, China
3. Nanjing Medical University, Nanjing 210029, Jiangsu, China
*Corresponding author.) |
| 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. |
| Key words: pioglitazone pancreatitis monocytes NF-κB p38 mitogen-activated protein kinases |
