肠复方对葡聚糖硫酸钠诱导的急性溃疡性结肠炎小鼠的干预作用

Decoction for intestinal recovery alleviates acute ulcerative colitis induced by dextran sulfate sodium in mice

目的观察肠复方对葡聚糖硫酸钠诱导的急性溃疡性结肠炎(UC)小鼠症状、结肠组织炎症程度的影响。方法将40只雄性C57BL/6小鼠随机分为空白组、模型组、肠复方组和美沙拉秦组,每组10只。除空白组外,其余各组均自由饮用3%葡聚糖硫酸钠7 d建立急性UC模型。实验第8天开始,肠复方组给予0.85 g/mL的肠复方颗粒溶液灌胃,美沙拉秦组给予0.08 g/mL的美沙拉秦缓释颗粒混悬液灌胃,空白组及模型组给予等体积蒸馏水灌胃,均1次/d,连续7 d。实验过程中每天记录小鼠一般情况并计算疾病活动指数(DAI)。实验第15天取材,测量结肠长度,苏木精-伊红染色法观察结肠组织病理形态并评分,ELISA法检测结肠组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-4(IL-4)、白细胞介素-10(IL-10)水平,Western blot法检测结肠组织中闭合小环蛋白-1(ZO-1)、闭合蛋白-1(Claudin-1)表达情况。结果与空白组比较,模型组小鼠结肠长度明显缩短(P<0.05),DAI、结肠组织病理评分及结肠组织中TNF-α、IL-6水平均明显升高(P均<0.05),结肠组织中IL-4、IL-10水平均明显降低(P均<0.05),结肠组织中ZO-1、Claudin-1蛋白相对表达量均无明显变化(P均>0.05)。肠复方组与美沙拉秦组小鼠结肠长度均明显长于模型组(P均<0.05),DAI、结肠组织病理评分及结肠组织中TNF-α、IL-6水平均明显低于模型组(P均<0.05),结肠组织中IL-4、IL-10水平及结肠组织中ZO-1、Claudin-1蛋白相对表达量与模型组比较差异均无统计学意义(P均>0.05),且肠复方组与美沙拉秦组各指标比较差异均无统计学意义(P均>0.05)。结论肠复方可调节由葡聚糖硫酸钠诱导的急性UC小鼠的肠黏膜炎症因子水平,改善小鼠症状及减轻结肠组织炎症,其作用与美沙拉秦缓释颗粒等效。

Objective It is to observe the effects of decoction for intestinal recovery on symptoms and inflammation of colonic tissue in mice with acute ulcerative colitis(UC)induced by dextran sulfate sodium(DSS).Methods Forty male C57BL/6 mice were randomly divided into blank group,model group,decoction for intestinal recovery group and mesalazine group,with 10 mice in each group.The mice of all groups except for the blank group were gavaged with 3%DSS for 7 days to establish acute UC models.On the 8th day of the experiment,the decoction for intestinal recovery group was given granule solution of decoction for intestinal recovery 0.85 g/mL by gavage,the mesalazine group was given mesalazine sustained-release granule suspension 0.08 g/mL by gavage,while the normal group and the model group were given an equal volume of distilled water by gavage,all once daily,continuously treated for 7 days.During the experiment,the general condition of the mice was recorded daily and their disease activity index(DAI)was calculated.On the 15 th day of the experiment,the length of colon was measured,the pathological morphology of colon tissue was observed and scored by hematoxylin-eosin staining,and the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-4(IL-4),interleukin-10(IL-10)in colon tissue were detected by enzyme-linked immunosorbent assay(ELISA),the expressions of ZO-1 and CLAUDIN-1 proteins in colon tissue were detected by Western-Blot.Results Compared with the blank group,the colon length of mice in the model group was significantly shorter(P<0.05),the DAI,colon histopathological score and the levels of TNF-αand IL-6 in colon tissues were significantly higher(all P<0.05),the levels of IL-4 and IL-10 in colon tissues were significantly lower(all P<0.05),and the relative expressions of ZO-1 and Claudin-1 proteins in colon tissue were not significantly changed(all P>0.05).The colon length of mice in the decoction for intestinal recovery group and mesalazine group was significantly longer than that in the model group(all P<0.05),and the DAI,colon histopathological score and levels of TNF-αand IL-6 in colon tissue were significantly lower than those in the model group(all P<0.05),but the differences in the levels of IL-4 and IL-10 in colon tissue and the relative expressions of ZO-1 and Claudin-1 in colon tissue were not statistically significant compared with those in the model group(all P>0.05),the differences in each index were also not statistically significant between the decoction for intestinal recovery group and mesalazine group(all P>0.05).Conclusion Decoction for intestinal recovery can regulate the levels of inflammatory factors in intestinal mucosa,thus to improve the symptoms and relieve inflammation of colon tissue in mice with acute ulcerative colitis induced by DSS,its efficacy is equivalent to mesalazine sustained-release granules.