中药槐花对溃疡性结肠炎小鼠肠道黏膜通透性的实验研究

Experimental Study on Influence of Huaihua(Sophorae Flos)on Permeability of Intestinal Mucosa in Mice with Ulcerative Colitis

目的研究槐花对肠道黏膜通透性的影响。方法采用葡聚糖硫酸钠(DSS)建立小鼠溃疡性结肠炎模型,随机分为对照组、急性肠炎模型组、低浓度槐花给药组(低剂量1.04 g·kg^(-1))和高浓度槐花给药组(高剂量3.12 g·kg^(-1)),每组6只,除对照组小鼠给予蒸馏水饮用外,其他组每天自由饮用新鲜配制的3%DSS溶液连续饮用7 d以构建肠炎模型。低、高剂量槐花给药组分别通过灌胃不同浓度槐花悬液给药,对照组及模型组灌胃等体积生理盐水。给药期间,观察小鼠一般情况。给药结束后,对小鼠进行表型测定,采用苏木素-伊红(HE)染色法观察结肠组织病理形态并进行组织学评分,采用血清异硫氰酸荧光素(FITC-dextran)检测肠道黏膜屏障完整性,采用免疫荧光检测两种紧密连接蛋白包括闭锁小带蛋白1(ZO-1)、咬合蛋白(occludin)的表达水平。结果(1)与对照组比较,模型组小鼠体质量、结肠长度明显降低,脾重量、粪便含水量和疾病活动指数(DAI)评分显著升高;与模型组比较,低、高浓度槐花给药组小鼠体质量、结肠长度均显著升高,脾重量、粪便含水量和活动指数(DAI)评分均显著降低。(2)与对照组比较,模型组结肠组织病理学评分升高(P<0.0001);与模型组比较,低、高浓度槐花给药组结肠组织病理学评分均显著降低(P<0.05或P<0.0001)。(3)与对照组比较,模型组小鼠肠道黏膜通透性显著升高;与模型组比较,低、高浓度槐花给药组黏膜通透性显著降低。(4)与对照组比较,模型组小鼠两种紧密连接蛋白ZO-1以及occludin表达水平明显降低;与模型组比较,低、高浓度槐花给药组两种紧密连接蛋白的表达水平有所升高。结论槐花对小鼠肠道黏膜通透性有恢复作用,并可改善小鼠溃疡性结肠炎状况。

Objective To study the effect of Huaihua(Sophorae Flos)on the permeability of intestinal mucosa.Methods The ulcerative colitis model in mice was established with dextran sodium sulfate(DSS),and they were randomly divided into control group,acute enteritis model group,Huaihua(Sophorae Flos)low concentration administration group(1.04 g·kg^(-1))and Huaihua(Sophorae Flos)high concentration administration group(3.12 g·kg^(-1)),at least 6 mice in each group.The mice in the control group were given distilled water for drinking,and the mice of the other groups were free to drink freshly prepared 3%DSS solu-tion every day for continuous 7 days to construct the enteritis model.Huaihua(Sophorae Flos)low and high concentration admin-istration groups were administered different concentrations of Huaihua(Sophorae Flos)suspension by intragastric administration and the control group and model group were intragastrically administered the equal volumes of normal saline.During the adminis-tration period,the general condition of the mice was observed.After the administration,the phenotype of the mice was deter-mined,the pathological morphology of the colon tissue was observed by hematoxylin-eosin(HE)staining and histological scoring was performed,the integrity of the intestinal mucosal barrier was detected by serum FITC-dextran,and the integrity of the intesti-nal mucosal barrier was detected by immunofluorescence.The expression levels of two tight junction proteins ZO-1 and occludin were detected.Results(1)Compared with those of the control group,the body weight and colon length of the mice in the model group were significantly reduced,and the spleen weight,fecal water content and disease activity index(DAI)score were signifi-cantly increased.Compared with those of the model group,the body weight and colon length of the mice in the Huaihua(Sophorae Flos)low and high concentration groups were significantly increased,and the spleen weight,fecal water content and DAI score were all significantly decreased.(2)Compared with those of the control group,the histopathological score of the colon in the mod-el group increased(P<00001).Compared with those of the model group,the histopathological scores of the colon in the Hua-ihua(Sophorae Flos)low and high concentration groups were significantly reduced(P<0.05 or P<00001).(3)Compared with that of the control group,the intestinal mucosal permeability of the mice in the model group was significantly increased.Compared with that of the model group,the mucosal permeability of the Huaihua(Sophorae Flos)low and high concentration groups was significantly reduced.(4)Compared with those of the control group,the expression levels of two tight junction proteins ZO-1 and occludin in the model group were significantly lower.Compared with those of the model group,the expression levels of two tight junction proteins ZO-1 and occludin in the Huaihua(Sophorae Flos)low and high concentration groups were in-creased.Conclusion Huaihua(Sophorae Flos)can restore the permeability of intestinal mucosa in mice and improve the condition of ulcerative colitis in mice.