醋炙减弱甘遂醋酸乙酯部位对小鼠胃肠黏膜通透性影响的研究

Attenuation by stir baking with vinegar on effect of ethyl acetate fraction from Kansui Radix on mice gastrointestinal permeability

目的探讨醋炙减弱甘遂醋酸乙酯部位对小鼠胃肠道黏膜通透性影响的机制。方法将小鼠分为对照组、甘遂组、醋甘遂组,ig给药7 d后,取小鼠胃、肠组织进行透射电镜观察;并通过免疫组化的方法,测定胃肠组织中E-钙黏蛋白和血管细胞黏附分子-1(VCAM-1)蛋白的表达。结果透射电镜观察结果显示,甘遂组胃固有腺体上皮排列紊乱,细胞质电子密度不均匀,细胞核形态不规则,腺上皮表面有大量溶酶体出现;醋甘遂组胃固有形态上皮细胞形态规则,部分线粒体畸形;甘遂组肠黏膜表面微绒毛高度变矮,细胞间连接丰富,但近表面的紧密连接较短;醋甘遂组肠上皮细胞排列规则,细胞形态一致,表面微绒毛数量减少、高度变矮,但较规则,线粒体结构良好。免疫组化检查结果显示,与对照组相比,甘遂组E-钙黏蛋白表达降低,VCAM-1蛋白表达增强(P<0.05、0.01);与甘遂组相比,醋甘遂组E-钙黏蛋白表达增强(P<0.05、0.01),VCAM-1表达降低(P<0.05、0.01)。结论醋炙能明显降低甘遂对小鼠胃肠黏膜上皮细胞的刺激,减弱对胃肠黏膜通透性的影响,从而降低甘遂的刺激性。

Objective To study the mechanism of attenuation by stir baking with vinegar on effect of ethyl acetate fraction of Kansui Radix on mice gastrointestinal permeability. Methods Mice were randomly divided into blank control, Kansui Radix, and Kansui Radix stir-baked with vinegar groups. The gastric and intestinal tissues were removed from mice after ig administration for 7 d and were observed by the transmission electron microscope （TEM）. The expression of E-cadherin and VCAM-1 was examined by immunohistochemical techniques. Results Using TEM, the arrangement and karyon morphology of gastric glandular epithelium cells were irregular with the uneven electronic of cytoplasm and lots of lysosomes in Kansui Radix group; The morphology of the gastric epithelial cells was irregular with some abnormal mitochondrions in the cytoplasm in Kansui Radix stir-baked with vinegar group; The intestinal mucosa cells were covered with short microvilli with well-connected intercellular junctions, but the tight junctions on the near-surface were short in Kansui Radix group; The arrangement and morphology of intestinal epithelial cells were fairly uniform with few and short microvilli and intact mitochondrion in Kansui Radix stir-baked with vinegar group. Using immunohistochemical techniques, compared with the control group, Kansui Radix could decrease the expression of E-cadherin and increase the expression of VCAM-1 （P 〈 0.05, 0.01）. While compared with Kansui Radix group, Kansui Radix stir-baked with vinegar could obviously increase the expression ofE-cadherin （P 〈 0.05, 0.01） and decrease the expression ofVCAM-I （P 〈 0.05, 0.01）. Conclusion Stir-baking with vinegar can attenuate the Kansui Radix-induced gastrointestinal inflammation. The mechanism may be that it can improve the morphology of mice gastrointestinal mucosal cells, reduce the membrane permeability, and decrease the stimulating effect of Kansui Radix.