过氧化物酶体增殖物激活受体γ激动剂对大鼠胰腺腺泡细胞信号传导途径的影响  

The effect of peroxisome proliferator-activated receptor gamma ligand on signaling pathway in pancreatic acinar cells

作　　者：万红宇[1] 袁耀宗[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院消化科,200025

出　　处：《中华消化杂志》2010年第1期42-46,共5页Chinese Journal of Digestion

摘　　要：目的了解经雨蛙肽处理的胰腺腺泡细胞AR42J中过氧化物酶体增殖因子活化受体γ（PPARγ）与核因子（NF）－κB活性间的关系。方法将胰腺腺泡AR42J细胞分为对照组（常规培养）、吡咯列酮组（40μmol／L吡咯列酮）、吡咯列酮＋雨蛙肽组（40μmol／L吡咯列酮＋10^-8mol／L-雨蛙肽）、雨蛙肽组（40μmol／L二甲基亚砜10^-8mol／L雨蛙肽）和吡咯列酮＋GW9662＋雨蛙肽组（40μmol／L吡咯列酮＋5μmol／L GW9662＋10^-8mol／L雨蛙肽），各组培养30min后检测PPART和NF—κB活性。Western印迹法检测NFκB、PPARy蛋白和磷酸化NFκB抑制物（IκB）α抗体、IκB激酶（IKK）β和IκB0的表达差异、IKKβ活性和IκBα磷酸化的变化。免疫荧光法和Western印迹法检测NF—κB（p65和p50）的核移位。免疫沉淀法检测IκBα与NF-κB的变化。结果吡咯列酮不仅抑制IKKβ活性（吡咯列酮＋雨蛙肽组：雨蛙肽组为1．6：3．7）及IκBα的磷酸化（吡咯列酮＋雨蛙肽组：雨蛙肽组为0．9：1．5），还能加强IκBα与NF—κB的结合（吡咯列酮＋雨蛙肽组：雨蛙肽组为0．8：0．3），抑制NFκB的核移位及NF—κB的转录活性（P〈0．01），而PPARγ拮抗剂GW9662则逆转了吡咯列酮对NF-κB活性的抑制作用（P〈0．05）。结论在雨蛙肽处理的AR42J细胞中PPAR7通过干扰NF—κB的活化产生抗炎作用。Objective To investigate the putative relationship between peroxisome proliferators activated receptor gamma （PPAR7） and nuclear factor （NF）-κB in cerulein-treated pancreatic aeinar AR42J cells. Methods The AR42J ceils were allocated to control group, pioglitazone group （treated with 40 μmol/L of pioglitazone）, pioglitazone ＋ cerulein group （treated with 40 μmol/L of pioglitazone＋ 10 8 rnol/L of cerulein） and pioglitazone ＋ cerulein＋ PPARγ antagonist （GW9662） group （treated with 40 μmol/L of pioglitazone＋ 5 μmol/L of GW9662 ＋ 10^- 8 mol/L of eerulein）. Activity of NF-κB and PPARy expression were detected 30 minuts after stimulated by cerulein with or without the presence of pioglitazone. The protein expressions of NF-κB and PPARγ, antibody to IκBα phosphorylation, the differential expression between IκB kinase （IKK）β and IκBa, the IKKJ3 activity as well as changes of pIκBa were examined by Western blotting. The nuclear accumulation of NF κB （p65 and p50 subunits） was determined by immunofluorescence and Western blotting. The interaction between NF-κB p65 and IκBa was observed by immunoprecitation. Results Treatment of AR42J cells with pioglitazone attenuated cerulein induced cytosolic activity of IKK protein （1. 6 ： 3. 7）or IκBα phosphorylation （0.9 ： 1. ,5） , strengthened the integration of IκBα and NF κB （0.8 ： 0.3） , inhibited transcription activity of p50 and p65 NF-κB dimer and nuclear accumulation （P〈0.01）. Adversely, the inhibitory effect of pioglitazone on NF -κB activity induced by cerulein was almost reversed by GW9662 （P〈0.05）. Conclusion These findings provide evidence for the involvement of PPARy in the activity of NF -κB in cerulein treated AR42J ceils.

关 键 词：炎症 雨蛙肽 核因子-κB 过氧化物酶体增殖物激活受体 吡咯列酮 

分 类 号：R965[医药卫生—药理学]