摘要
目的:应用HPLC法测定结肠中γ-氨基丁酸(GABA)含量,并用免疫组化法检测GABA受体的表达,探讨GABA在溃疡性结肠炎发病中的作用。方法:雄性Wistar大鼠16只随机分为对照组、实验组,对照组自由饮用蒸馏水,实验组自由饮用含3%DSS的蒸馏水7d以诱导建立大鼠溃疡性结肠炎模型,1周后处死所有动物,分离结肠,结肠组织匀浆,应用HPLC法测定结肠组织中GABA含量;HE染色镜下观察病理变化,免疫组化法SP法检测病变组织GABAA受体、GABAB受体的表达水平。结果:实验组大鼠大肠组织内GABA浓度为(698.6±23.5)ng/ml,对照组大鼠大肠组织内GABA浓度为(220±15.9)ng/ml,实验组GABA含量较正常组升高明显,差异具有统计学意义(P<0.05)。镜下实验组以及对照组均能看到阳性表达细胞,受体染色主要在细胞膜及胞浆。与对照组相比,实验组大肠镜下GABA受体阳性细胞明显增多,其差异具有统计学意义(P<0.05))。结论:溃疡性结肠炎的发生发展与结肠组织GABA的增高有关,GABA主要通过其GABAA受体参与溃疡性结肠炎的病理生理过程。其可能机制尚需做进一步的研究。
Objective:Use the way of HPLC to determinate the content of GABA in the colonic,and test the expression of the receptor of the GABA with the immunohistochemical method,and study the effect of GABA in the ulcerative colitis.Methods:Modeling uncreative colitis in rats which induced by sixteen male Wistar rats were randomly divided into control group and experimental group,and the control group drank the distilled water freely,but the experimental group drank the distilled water with 3%DSS,after seven days,killed all the rats,and separated their colon,and homogenated the tissues to use the way of HPLC to determinate the content of GABA in the colon;after HE staining to observe the pathologic change under microscope,and use the way of SP of the immunohistochemical method to test the expression level of the receptor of the GABAA and GABAB from the lesion tissues.Results:The concentration of GABA in the colonic tissues of the experimental group was(698.6 ± 23.5)ng/ml and the control group was(220±15.9)ng/ml,so the content of GABA in the colon of the experimental group was higher than the normal group,and the difference had significance of statistics(P<0.05).Both the experimental group and the control group could see the positive expression cells,the staining of the receptor was mainly at cell membrane and cytoplasm.Comparing with the control group,there was an evident increase of the positive expression cells of the receptor of the GABA in the experimental group under microscope,and the difference had significance of statistics s(P<0.05).Conclusion:The occurrence and development of the uncreative colitis related to the increase of the GABA in the colonic tissues,and mainly by the receptor of GABAA,the GABA participate the pathophysiological process of the uncreative colitis,but it need a further research about the possible mechanism.
基金
湖北省自然科学基金资助项目(2010CDB04405)
