肾上腺髓质素对肠上皮细胞缺氧/复氧损伤后屏障功能的影响  

Effect of adrenomedullin on intestinal epithelial barrier function under hypoxia/reoxygenation injury and possible mechanism

作　　者：田铸[1] 周维均 苟亚军[1] 

机构地区：[1]第三军医大学西南医院急救部 [2]解放军77113部队卫生队

出　　处：《局解手术学杂志》2016年第4期251-254,共4页Journal of Regional Anatomy and Operative Surgery

摘　　要：目的研究肾上腺髓质素(AM)在缺氧/复氧(H/R)条件下对肠上皮细胞屏障功能的影响,对其机制进行了初步探讨。方法建立单层肠上皮Caco-2细胞缺氧/复氧损伤模型,实验分为3组:正常细胞培养(N)组、缺氧/复氧损伤模型(H/R)组、缺氧/复氧损伤+肾上腺髓质素(H/R+AM)组。检测跨上皮细胞电阻(TEER)值,使用激光共聚焦显微镜观察紧密连接的变化,Western blot方法检测细胞紧密连接蛋白Occludin和ZO-1的表达及核因子-κB(NF-κBp65)蛋白表达水平。结果与H/R组相比,H/R+AM组的紧密连接破坏明显减少;N组,H/R组和H/R+AM组的TEER值分别为(157.68±5.54)、(96.06±3.42)、(134.56±4.72)Ohm/cm2,H/R+AM组较H/R组升高了40.00%(P<0.05)。N组,H/R组和H/R+AM组的Occludin和ZO-1蛋白表达量分别为(0.43±0.03)和(0.27±0.04)、(0.20±0.03)和(0.15±0.07)、(0.32±0.15)和(0.21±0.03),H/R+AM组的Occludin和ZO-1蛋白表达量较H/R组分别升高60.00%、40.00%(P<0.05),N组、H/R组、H/R+AM组的NF-κBp65蛋白表达量分别为(0.53±0.30)、(2.89±0.16)、(1.75±0.25),H/R+AM组的NF-κBp65蛋白表达量较H/R组降低了40.00%。结论 AM可通过减少H/R损伤诱导的紧密连接的破坏来保护肠上皮屏障,其机制可能与抑制NF-κB信号通路有关。Objective To investigate the effect of adrenomedullin on epithelial barrier function of Caco-2 cells under hypoxia / reoxygenation injury and the molecular mechanism. Methods Build hypoxia / reoxygenation injury model,Caco-2 cells were randomly divided into three groups： normal control group（ N）,hypoxia / reoxygenation group（ H / R） and hypoxia / reoxygenation plus 1 μmol / L adrenomedullin group（ H / R ＋ AM）. Transepithelial electrical resistance（ TEER） was determined,and the laser confocal microscope was used to detect the construction of tight junction. The protein expressions of tight junction proteins（ Occludin and ZO-1） and nuclear factor kappa B（ NF-κBp65）were examined by using western blotting. Results Pretreatment of AM significantly attenuated the disruption of morphological structure of tight junction caused by H / R. The TEER of N group,H / R group and H / R ＋ AM group were（ 157. 68 ± 5. 54）,（ 96. 06 ± 3. 42）,（ 134. 56 ±4. 72） Ohm / cm2 respectively. After AM pretreatment,the TEER were significantly raised compared with that in H / R group by 40. 00%（ P〈0. 05）. The relative protein expressions of Occludin and ZO-1 in N group,H/R group and H/R ＋ AM group were [（ 0. 43 ± 0. 03）,（ 0. 27 ± 0. 04） ],[（ 0. 20 ± 0. 03）,（ 0. 15 ± 0. 07） ],[（ 0. 32 ± 0. 15）,（ 0. 21 ± 0. 03） ]respectively. After AM pretreatment,the protein expression of Occludin and ZO-1 were significantly raised compared with that in H / R group（ by 60. 00% and 40. 00%）. Simultaneously,the relative protein expressions of NF-κBp65 in N group,H / R group and H / R ＋ AM group were（ 0. 53 ± 0. 30）,（ 2. 89 ± 0. 16）,（ 1. 75 ± 0. 25）respectively. And H / R induced NF-κBp65 activation was significantly inhibited by AM treatment（ decreased as compared with H / R group by40. 00%）. Conclusion AM can attenuate intestinal epithelial barrier dysfunction by regulating the expression of intestinal tight junction proteins. The inhibition of NF-κB ac

关 键 词：肾上腺髓质素 缺氧/复氧 肠上皮屏障 紧密连接蛋白 

分 类 号：R333.3[医药卫生—人体生理学]