中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2010年
1期
42-46
,共5页
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
炎癥%雨蛙肽%覈因子-κB%過氧化物酶體增殖物激活受體%吡咯列酮
염증%우와태%핵인자-κB%과양화물매체증식물격활수체%필각렬동
Inflammation%Cerulein%Nuclear factor-κB%Peroxisome proliferator-activated receptors%Pioglitazone
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(40 μmol/L吡咯列酮)、吡咯列酮+雨蛙肽组(40 μmol/L吡咯列酮+10~(-8) mol/L雨蛙肽)、雨蛙肽组(40 μmol/L二甲基哑砜+10~(-8)mol/L雨蛙肽)和吡咯列酮+GW9662+雨蛙肽组(40 μmol/L吡咯列酮+5 μmol/L GW9662+10~(-8)mol/L雨蛙肽),各组培养30 min后检测PPARγ和NF-κB活性.Western印迹法检测NF-κB、PPARγ蛋白和磷酸化NF-κB抑制物(IκB)α抗体、IκB激酶(IKK)β和IκBα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+雨蚌肽组: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NF-κB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PPARγ通过干扰NF-κB的活化产生抗炎作用.
目的 瞭解經雨蛙肽處理的胰腺腺泡細胞AR42J中過氧化物酶體增殖因子活化受體γ(PPARγ)與覈因子(NF)-κB活性間的關繫.方法 將胰腺腺泡AR42J細胞分為對照組(常規培養)、吡咯列酮組(40 μmol/L吡咯列酮)、吡咯列酮+雨蛙肽組(40 μmol/L吡咯列酮+10~(-8) mol/L雨蛙肽)、雨蛙肽組(40 μmol/L二甲基啞砜+10~(-8)mol/L雨蛙肽)和吡咯列酮+GW9662+雨蛙肽組(40 μmol/L吡咯列酮+5 μmol/L GW9662+10~(-8)mol/L雨蛙肽),各組培養30 min後檢測PPARγ和NF-κB活性.Western印跡法檢測NF-κB、PPARγ蛋白和燐痠化NF-κB抑製物(IκB)α抗體、IκB激酶(IKK)β和IκBα的錶達差異、IKKβ活性和IκBα燐痠化的變化.免疫熒光法和Western印跡法檢測NF-κB(p65和p50)的覈移位.免疫沉澱法檢測IκBα與NF-κB的變化.結果 吡咯列酮不僅抑製IKKβ活性(吡咯列酮+雨蛙肽組:雨蛙肽組為1.6;3.7)及IκBα的燐痠化(吡咯列酮+雨蚌肽組:雨蛙肽組為0.9:1.5),還能加彊IκBα與NF-κB的結閤(吡咯列酮+雨蛙肽組:雨蛙肽組為0.8:0.3),抑製NF-κB的覈移位及NF-κB的轉錄活性(P<0.01),而PPARγ拮抗劑GW9662則逆轉瞭吡咯列酮對NF-κB活性的抑製作用(P<0.05).結論 在雨蛙肽處理的AR42J細胞中PPARγ通過榦擾NF-κB的活化產生抗炎作用.
목적 료해경우와태처리적이선선포세포AR42J중과양화물매체증식인자활화수체γ(PPARγ)여핵인자(NF)-κB활성간적관계.방법 장이선선포AR42J세포분위대조조(상규배양)、필각렬동조(40 μmol/L필각렬동)、필각렬동+우와태조(40 μmol/L필각렬동+10~(-8) mol/L우와태)、우와태조(40 μmol/L이갑기아풍+10~(-8)mol/L우와태)화필각렬동+GW9662+우와태조(40 μmol/L필각렬동+5 μmol/L GW9662+10~(-8)mol/L우와태),각조배양30 min후검측PPARγ화NF-κB활성.Western인적법검측NF-κB、PPARγ단백화린산화NF-κB억제물(IκB)α항체、IκB격매(IKK)β화IκBα적표체차이、IKKβ활성화IκBα린산화적변화.면역형광법화Western인적법검측NF-κB(p65화p50)적핵이위.면역침정법검측IκBα여NF-κB적변화.결과 필각렬동불부억제IKKβ활성(필각렬동+우와태조:우와태조위1.6;3.7)급IκBα적린산화(필각렬동+우방태조:우와태조위0.9:1.5),환능가강IκBα여NF-κB적결합(필각렬동+우와태조:우와태조위0.8:0.3),억제NF-κB적핵이위급NF-κB적전록활성(P<0.01),이PPARγ길항제GW9662칙역전료필각렬동대NF-κB활성적억제작용(P<0.05).결론 재우와태처리적AR42J세포중PPARγ통과간우NF-κB적활화산생항염작용.
Objective To investigate the putative relationship between peroxisome proliferators activated receptor gamma (PPARγ) and nuclear factor (NF)-κB in cerulein-treated pancreatic aeinar AR42J cells. Methods The AR42J cells 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) mol/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 cerulein). Activity of NF-κB and PPARγ 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 IKKβ 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 IκBα was observed by immunopreeitation. Results Treatment of AR42J cells with pioglitazone attenuated cerulein induced cytosolic activity of IKK protein (1.6 : 3.7)or IκBa 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 PPARγ in the activity of NF-κB in cerulein treated AR42J cells.
