CAJ | 학술논문

目的观察骨桥蛋白(osteopontin,OPN)在右旋葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的小鼠结肠炎中的变化,探讨OPN在炎症性肠病(inflammatory bowel disease,IBD)发病中的作用.方法 32只雄性BALB/C小鼠随机分为对照组(予蒸馏水饮用23 d)、模型组(5%DSS饮用9 d后2.5%DSS维持2周)、柳氮磺胺吡啶(SASP)治疗组(在模型组基础上从第10天起予SASP 600 mg/kg灌胃2周)和英夫利昔治疗组(在模型组基础上在第10天予尾静脉注射英夫利昔100 mg/kg 1次).酶联免疫法检测小鼠血浆OPN浓度,逆转录聚合酶链反应和Western印迹法检测小鼠结肠组织中OPN的mRNA水平和蛋白质水平表达,免疫组化法检测OPN在结肠组织中的定位表达.结果各组小鼠的血浆OPN浓度分别为(5.26±1.93)、(10.21±2.37)、(4.58±1.83)和(4.82±1.83)ng/ml;结肠组织中OPN mRNA表达量分别为(0.36±0.16)、(0.71±0.17)、(0.32±0.07)和(0.42±0.22);结肠组织中OPN的蛋白表达量分别为(0.44±0.10)、(0.85±0.04)、(0.61±0.11)和(0.58±0.17);结肠黏膜固有层OPN阳性细胞数分别为(46.6±10.9)、(155.5±43.8)、(73.1±6.8)和(70.6±8.3).与对照组相比,模型组血浆和结肠组织中OPN的表达明显增高(P均<0.05),SASP治疗组和英夫利昔治疗组OPN的表达均较SASP治疗组下降(P均<0.05),SASP治疗组和英夫利昔治疗组OPN的表达差异无统计学意义.结论 OPN在DSS诱导的小鼠结肠炎中表达增加,经药物治疗后表达降低.OPN促进了DSS诱导的小鼠结肠炎的发生.
목적 관찰골교단백(osteopontin,OPN)재우선포취당류산납(dextran sodium sulfate,DSS)유도적소서결장염중적변화,탐토OPN재염증성장병(inflammatory bowel disease,IBD)발병중적작용.방법 32지웅성BALB/C소서수궤분위대조조(여증류수음용23 d)、모형조(5%DSS음용9 d후2.5%DSS유지2주)、류담광알필정(SASP)치료조(재모형조기출상종제10천기여SASP 600 mg/kg관위2주)화영부리석치료조(재모형조기출상재제10천여미정맥주사영부리석100 mg/kg 1차).매련면역법검측소서혈장OPN농도,역전록취합매련반응화Western인적법검측소서결장조직중OPN적mRNA수평화단백질수평표체,면역조화법검측OPN재결장조직중적정위표체.결과 각조소서적혈장OPN농도분별위(5.26±1.93)、(10.21±2.37)、(4.58±1.83)화(4.82±1.83)ng/ml;결장조직중OPN mRNA표체량분별위(0.36±0.16)、(0.71±0.17)、(0.32±0.07)화(0.42±0.22);결장조직중OPN적단백표체량분별위(0.44±0.10)、(0.85±0.04)、(0.61±0.11)화(0.58±0.17);결장점막고유층OPN양성세포수분별위(46.6±10.9)、(155.5±43.8)、(73.1±6.8)화(70.6±8.3).여대조조상비,모형조혈장화결장조직중OPN적표체명현증고(P균<0.05),SASP치료조화영부리석치료조OPN적표체균교SASP치료조하강(P균<0.05),SASP치료조화영부리석치료조OPN적표체차이무통계학의의.결론 OPN재DSS유도적소서결장염중표체증가,경약물치료후표체강저.OPN촉진료DSS유도적소서결장염적발생.
Objective To investigate the changes of osteopontin (OPN) expression in murine colitis induced by dextran sodium sulfate (DSS) and its role in inflammatory bowel disease (IBD).Methods Thirty-two male BALB/C mice were randomly assigned into 4 groups: control group (group A), DSS-induced murine colitis model group (group B), salazosulfapyridine treatment group (group C) and infliximab treatment group (group D). Plasma OPN concentration was measured by enzyme linked immunosorbent assay (ELISA). OPN mRNA level was detect by using reverse transcriptase polymerase chain reaction (RT-PCR) and protein expression in colonic tissues was analyzed by using Western blotting. The location expression of OPN in colonic tissues was determined by immunohistochemical staining. Results In group A, B,C and D, the plasma concentrations of OPN were (5.26±1.93) ng/ml, (10.21±2.37) ng/ml, (4.58±1.83) ng/ml and (4.82±1.83) ng/ml,respectively: the mRNA levels of OPN in colonic tissues were (0.36±0.16), (0.71±0.17),(0.32±0.07) and (0. 34±0. 08), respectively; the protein levels of OPN in colonic tissues were (0.44±0.10), (0.85±0.04), (0.61±0.11) and (0.58±0.17), respectively. The OPN-positive cell numbers in lamina propria mononuclear were (46.6±10.9), (155.5±43.8), (73.1±6.8) and (70.6±8.3), respectively. The expression of OPN in group B was significantly higher than that in group A (P<0.05). Compared with group B, the expressions of OPN in group C and D were significantly decreased (all P valus <0. 05). No significant difference was detected between group C and D. Conclusions The study shows that the expression of OPN significantly increased in DSS-induced murine colitis and decreased after treated with drugs. OPN-mediated immune response contributes to DSS-induced murine colitis.