阻塞性黄疸对肠上皮细胞氯离子分泌影响的研究  

Effects of obstructive jaundice on enterocyte chloridion secretion in rats

作　　者：陈振勇[1] 戴红梅[1] 杨鹏[1] 王延刚[1] 黄文广[1] 冯贤松[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院综合外科,湖北武汉430022

出　　处：《中国病理生理杂志》2011年第11期2194-2198,共5页Chinese Journal of Pathophysiology

基　　金：湖北省自然科学基金资助项目(No.2010CDB08005)

摘　　要：目的:探讨阻塞性黄疸(OJ)对肠上皮细胞氯离子分泌的影响。方法:建立OJ大鼠模型,分别于胆管结扎7 d(OJ1组)和14 d(OJ2组)后,检测实验动物外周血Cl-浓度,取末端回肠上皮细胞体外培养,荧光分光光度法测量细胞内Cl-浓度,全细胞膜片钳技术检测Cl-电流的改变,并用Western blotting法分析电压门控氯离子通道蛋白2(ClC-2)表达的变化。结果:OJ1组和OJ2组的血清Cl-浓度分别为(88.67±4.68)mmol/L和(82.01±6.25)mmol/L,均低于对照组(均P<0.05)。细胞内Cl-相对浓度,OJ1组(3.14±0.38)和OJ2组(3.55±0.47)均高于对照组(均P<0.05)。对照组肠上皮细胞Cl-电流,从-80 mV的(-15.45±7.56)pA/pF升到80 mV的(5.85±0.81)pA/pF,呈外向整流趋势。OJ组Cl-平均电流密度绝对值逐渐减小,与对照组比较差异显著(均P<0.05)。Western blotting分析显示ClC-2蛋白表达条带在90 kD附近,对条带的定量分析表明,OJ1组为0.20±0.04,OJ2组为0.19±0.06,均低于对照组(0.27±0.06)(均P<0.05),但OJ1、2组间比较没有显著差异(P>0.05)。结论:OJ抑制肠腔氯离子分泌,缩小上皮细胞内外氯离子浓度差,减少Cl-外向电流,此作用与细胞膜上氯离子通道蛋白2的表达降低有关。AIM： To investigate the effects of obstructive jaundice（OJ） on enterocyte chloridion secretion in rats.METHODS： The OJ model of rats was set up.The rats were divided into OJ1 group and OJ2 group randomly.The animals in OJ1 group were sacrificed by exsanguination 7 days after operation and the rats in OJ2 group were sacrificed 14 days after operation.The Cl-concentration in peripheral blood was detected.The epithelial cells in the terminal ileum mucosas were cultured in vitro.The concentration of intracellular Cl-was detected by fluorospectrophotometry.The Cl-current was measured by whole-cell patch clamp experiments.Western blotting was used to examine the change of voltage-gated Cl-channel 2 protein（ClC-2） and the images were analyzed by image analysis system and statistical software quantitatively.RESULTS： The serum Cl-concentration in OJ1 group and OJ2 group were obviously lower than that in control group [（88.67±4.68） mmol/L and（82.01±6.25） mmol/L vs（95.57±7.51） mmol/L,both P0.05].The intracellular Cl-relative concentrations in OJ1 group and OJ2 group were higher than that in control group（3.14±0.38 and 3.55±0.47 vs 2.69±0.41,both P0.05）.The Cl-current of normal epithelial cells displayed outward rectification,and transformed negative value into positive value following cell membrane depolarization.The Cl-current was（-15.45±7.56） pA/pF and（5.85±0.81） pA/pF when voltage was-80 mV and 80 mV.The ranges of upgrade of absolute values of average electric current density in OJ groups were lower than those in control group（all P0.05）.Western blotting analysis showed that the ClC-2 protein generated a broad band about 90 kD.The average gradations of Western blotting images were lower in OJ groups than that in control group（0.20±0.04 and 0.19±0.06 vs 0.27±0.06,both P0.05）.However,no difference between the 2 OJ groups was observed（P0.05）.CONCLUSION： The chloridion secretion of intestinal epithelium is restrained in rats with OJ.The concentration gradient bet

关 键 词：黄疸 梗塞性 肠细胞 氯化物通道 

分 类 号：R364.5[医药卫生—病理学]