Resta-Lenert Silvia, Smitham Jane, Barrett Kim E
Division of Gastroenterology, Department of Medicine, University of California, School of Medicine, San Diego, CA 92103, USA.
Am J Physiol Gastrointest Liver Physiol. 2005 Jul;289(1):G153-62. doi: 10.1152/ajpgi.00395.2004. Epub 2005 Mar 17.
P-glycoprotein, the product of the multidrug resistance protein 1 (MDR1) gene, is a xenobiotic transporter that may contribute to the physiology of the intestinal barrier. Twenty-five percent of mdr1a-deficient (mdr1a(-/-)) mice spontaneously develop colitis at variable ages when maintained under specific pathogen-free conditions. We hypothesized that this disease would result from epithelial dysfunction and that conventional housing would increase incidence and severity of the colitis phenotype. Wild-type congenic FVB (+/+) mice were maintained under the same conditions as controls. Knockout and wild-type mice were matched for age and gender and observed for signs of colitis. Colonic tissues from both groups of mice were examined for macroscopic and microscopic injury and for basal ion transport and transepithelial resistance (TER). Translocation of bacteria across the intestine was assessed by culturing the spleen and mesenteric lymph nodes. Protein analysis was performed by Western blot analysis. All mdr1a(-/-) mice developed weight loss and signs of colitis, whereas wild-type mice never showed such signs. Within the mdr1a(-/-) group, males consistently developed severe colitis earlier than females. Knockout mice showed increased basal colonic ion transport (females, 162.7 +/- 4.6 vs. 49.7 +/- 3.8 muA/cm(2); males, 172.6 +/- 5.6 vs. 54.2 +/- 3.1 muA/cm(2); P < 0.01) and decreased TER (females, 25.4 +/- 0.3 vs. 36.4 +/- 0.8 Omega.cm(2); males, 23.1 +/- 1.0 vs. 38.3 +/- 0.2 Omega.cm(2); P < 0.01) compared with wild-type mice. Barrier dysfunction was accompanied by decreased phosphorylation of tight junction proteins. Expression of cyclooxygenase-2 and inducible nitric oxide synthase in intestinal tissues was increased in the mdr1a(-/-) group (P < 0.01) and correlated with disease severity. Bacterial translocation was greater both in incidence (P < 0.01) and severity (P < 0.001) for the knockout group. With respect to all indexes studied, mdr1a(-/-) males performed worse than females. Our data support the hypothesis that alterations in the intestinal barrier alone, in the absence of immune dysfunction, may rapidly lead to colitis in the setting of a normal colonic flora.
P-糖蛋白是多药耐药蛋白1(MDR1)基因的产物,是一种外源性物质转运蛋白,可能对肠道屏障的生理功能有作用。25%的多药耐药蛋白1a缺陷(mdr1a(-/-))小鼠在无特定病原体条件下饲养时,会在不同年龄自发发生结肠炎。我们推测这种疾病是由上皮功能障碍引起的,常规饲养会增加结肠炎表型的发生率和严重程度。野生型同基因FVB(+/+)小鼠与对照组在相同条件下饲养。基因敲除小鼠和野生型小鼠按年龄和性别匹配,观察结肠炎症状。检查两组小鼠的结肠组织,观察宏观和微观损伤情况以及基础离子转运和跨上皮电阻(TER)。通过培养脾脏和肠系膜淋巴结评估细菌在肠道内的移位情况。通过蛋白质印迹分析进行蛋白质分析。所有mdr1a(-/-)小鼠均出现体重减轻和结肠炎症状,而野生型小鼠从未出现此类症状。在mdr1a(-/-)组中,雄性小鼠比雌性小鼠更早出现严重结肠炎。与野生型小鼠相比,基因敲除小鼠的基础结肠离子转运增加(雌性:162.7±4.6对49.7±3.8 μA/cm²;雄性:172.6±5.6对54.2±3.1 μA/cm²;P<0.01),TER降低(雌性:25.4±0.3对36.4±0.8 Ω·cm²;雄性:23.1±1.0对38.3±0.2 Ω·cm²;P<0.01)。屏障功能障碍伴随着紧密连接蛋白磷酸化的减少。mdr1a(-/-)组肠道组织中环氧合酶-2和诱导型一氧化氮合酶的表达增加(P<0.01),且与疾病严重程度相关。基因敲除组的细菌移位在发生率(P<0.01)和严重程度(P<0.001)上都更高。就所有研究指标而言,mdr1a(-/-)雄性小鼠的表现比雌性小鼠更差。我们的数据支持这样的假设,即在没有免疫功能障碍的情况下,仅肠道屏障的改变就可能在正常结肠菌群环境中迅速导致结肠炎。