基于Th17/Treg免疫平衡观察清热化湿调枢方对溃疡性结肠炎模型小鼠的预防作用

Observation of Preventive Effect of Qingre Huashi Tiaoshu Recipe(清热化湿调枢方)on Ulcerative Colitis Model Mice Based on Th17/Treg Immune Balance

目的:基于Th17/Treg平衡的免疫调节作用观察清热化湿调枢方治疗溃疡性结肠炎(UC)的可能作用机制。方法:将60只C57BL/6小鼠随机分为正常组、模型组、清热化湿调枢方高剂量组、清热化湿调枢方中剂量组、清热化湿调枢方低剂量组、美沙拉嗪肠溶片组,每组10只。除正常组外,其余各组小鼠使用2.5%的葡聚糖硫酸钠(DSS)水溶液,自由饮用7 d复制UC模型,造模同时给予药物灌胃治疗。各给药组小鼠分别灌胃给予相应药物,正常组和模型组小鼠灌胃给予等体积纯净水。每天记录小鼠体质量、大便性状及便血情况。以结肠长度、结肠组织学评分反映肠道炎症水平,流式细胞术检测小鼠脾脏调节性T细胞(Treg)、辅助性T细胞17(Th17)百分比。结果:与正常组比较,模型组小鼠体质量明显减轻,结肠长度明显缩短,DAI评分明显升高,结肠黏膜腺体破坏程度及炎症细胞浸润程度较重,脾脏Treg细胞百分比及Treg/Th17值明显降低,Th17细胞百分比明显升高,差异均有统计学意义(P<0.05)。与模型组比较,清热化湿调枢方高、中剂量组小鼠结肠长度明显延长(P<0.05);清热化湿调枢方低剂量组小鼠结肠长度与模型组比较,差异无统计学意义(P>0.05)。与模型组比较,清热化湿调枢方高、中、低剂量组和美沙拉嗪肠溶片组小鼠体质量明显增加、DAI评分明显降低,且结肠黏膜腺体破坏程度及炎症细胞浸润程度减轻,脾脏Treg细胞百分比及Treg/Th17值明显升高,Th17细胞百分比明显降低,差异均有统计学意义(P<0.05)。结论:清热化湿调枢方可改善DSS诱导的溃疡性结肠炎小鼠腹泻、便血、体质量下降等症状和修复肠道黏膜损伤,其作用机制可能与调节Treg/Th17免疫失衡有关。

Objective:To observe the possible mechanism of Qingre Huashi Tiaoshu Recipe in the treatment of UC based on the immune regulation of Th17/Treg balance.Methods:60 C57BL/6 mice were randomly divided into normal group,model group,Qingre Huashi Tiaoshu Recipe high-dose group,Qingre Huashi Tiaoshu Recipe middle-dose group,Qingre Huashi Tiaoshu Recipe low-dose group,mesalazine enteric-coated tablets group,with 10 mice in each group.In addition to the normal group,the remaining groups of mice using 2.5%dextran sodium sulfate aqueous solution,free to drink 7 d copy UC model.Modeling while giving drug treatment.The mice in each administration group were given corresponding drugs by gavage,and the mice in the normal group and the model group were given equal volume of pure water by gavage.The body mass,stool characteristics and blood in stool were recorded every day.Colon length and histological score were used to reflect the level of intestinal inflammation.The percentage of spleen regulatory T cells(Treg)and helper T cells 17(Th17)in mice was detected by flow cytometry.Results:Compared with the normal group,the body mass of mice in the model group was significantly reduced,the length of colon was significantly shortened,the DAI score was significantly increased,the degree of damage to colonic mucosa glands and the degree of infiltration of inflammatory cells were heavier,the percentage of spleen Treg cells and Treg/Th17 value were significantly reduced,and the percentage of Th17 cells was significantly increased,with significant differences(P<0.05).Compared with the model group,the colon length of mice in the Qingre Huashi Tiaoshu Recipe high and middle-dose group of was significantly longer(P<0.05).There was no significant difference in colon length between the Qingre Huashi Tiaoshu Recipe low-dose group and the model group(P>0.05).Compared with the model group,the body mass of mice in the Qingre Huashi Tiaoshu Recipe high,middle and low-dose group and the mesalazine enteric-coated tablets group increased significantly,the DAI score decreased significantly,and the degree of damage to the colonic mucosa glands and the infiltration of inflammatory cells decreased,the percentage of spleen Treg cells and Treg/Th17 values increased significantly,and the percentage of Th17 cells decreased significantly(P<0.05).Conclusion:Qingre Huashi Tiaoshu Recipe can improve the symptoms of diarrhea,hematochezia and weight loss in DSS-induced ulcerative colitis mice and repair intestinal mucosal damage.Its mechanism may be related to the regulation of Treg/Th17 immune imbalance.