胃肠安丸对葡聚糖硫酸钠诱导溃疡性结肠炎小鼠的治疗作用及机制

目的探讨胃肠安丸对葡聚糖硫酸钠(DSS)诱导溃疡性结肠炎小鼠的治疗作用及机制。方法SPF级C57BL/6小鼠60只,除正常对照组小鼠外,其余小鼠均给予2.5%DSS水溶液,自由饮用7 d制备溃疡性结肠炎模型,模型评价后再随机分为模型组、美沙拉嗪组及胃肠安丸高剂量组、中剂量组、低剂量组。胃肠安丸低、中、高剂量组分别给予0.015、0.03、0.06 g/(kg·d)胃肠安丸混悬液0.5 mL灌胃,1次/d;美沙拉嗪组给予0.52 g/(kg·d)美沙拉嗪混悬液0.5 mL灌胃,1次/d;正常对照组和模型组给予等体积纯水灌胃,均连续干预10 d。对比各组小鼠疾病活动指数(DAI)评分、结肠长度,用HE染色观察小鼠结肠组织病理形态学变化,用流式细胞术检测脾组织中Treg、Th17、CD4^+、CD8^+细胞百分比,ELISA法检测血清细胞因子TNF-α、IL-6、IL-8、IL-10、CRP水平。结果药物干预过程中,各组小鼠体质量均逐日增加,腹泻、便血、体质量下降等症状逐日缓解,与模型组相比,美沙拉嗪组和胃肠安丸高剂量组能更快地缓解上述症状,2组DAI评分逐日下降(P<0.05)。药物干预结束后,与模型组比较,胃肠安丸高剂量组和美沙拉嗪组小鼠结肠较长(P<0.05),结肠组织充血、间质水肿及炎性细胞浸润较轻,CD4^+、CD8^+、Treg细胞百分比及血清IL-10水平较高(P<0.05),Th17细胞百分比及血清TNF-α、IL-6、IL-8、CRP水平较低(P<0.05)。结论胃肠安丸对DSS诱导的小鼠肠道黏膜损伤有一定的修复作用,其作用机制可能与恢复Treg/Th17免疫平衡以及调节相关细胞因子水平有关。

Therapeutic effect and mechanism of Weichang’an Pill on dextran sodium sulfate-induced ulcerative colitis in mice

Objective To explore the therapeutic effect and mechanism of Weichang’an Pill on dextran sodium sulfate(DSS)-induced ulcerative colitis in mice.Methods Sixty SPF C57 BL/6 mice were used.Except for normal control group of mice,the other mice were given 2.5%DSS aqueous solution with free consumption for 7 days to prepare ulcerative colitis models.After model evaluation,they were randomly divided into model group,Mesalazine group,Weichang’an Pill high-dose group,medium-dose group,low-dose group.The Weichang’an pill low,medium,and high dose groups were given 0.015,0.03,and 0.06 g/(kg·d)of Weichang’an pill suspension respectively by intragastric administration 0.5 mL,once a day;the mesalazine group was given 0.52 g/(kg·d)mesalazine suspension by intragastric administration of 0.5 mL once a day;the normal control group and model group were orally administered with equal volume of pure water,all for 10 consecutive days.The disease activity index(DAI)score and colon length of the mice in each group were compared,and the pathological and morphological changes of the colon tissue of the mice were observed with HE staining.The percentage of Treg,Th17,CD4^+,and CD8^+lymphocyte in the spleen tissue was detected by flow cytometry.Serum cytokines TNF-α,IL-6,IL-8,IL-10 and CRP were detected by ELISA.Results During the drug intervention,the body weight of the mice in each group was increased day by day,and the symptoms such as diarrhea,hematochezia,and weight loss were alleviated day by day.Compared with the model group,the above symptoms could be alleviated more rapidly in the mesalazine group and Weichang’an Pill high-dose group.DAI scores of both two groups were reduced day by day(P<0.05).After drug intervention,compared with the model group,the mice in the high-dose Weichang’an Pill group and mesalazine group had longer colons(P<0.05),and the congestion of colon tissue,interstitial edema,and inflammatory cell infiltration were milder.Compared with the model group,the percentage of CD4^+,CD8^+,and Treg cells and serum IL-10 levels in the high-dose group of Weichang’an Pill and mesalazine were higher(P<0.05);the percentage of Th17 cells and serum TNF-α,IL-6,IL-8,CRP level was lower(P<0.05).Conclusion Weichang’an Pill can alleviate the intestinal mucosal damage induced by DSS in mice.The mechanism may be related to the restoration of Treg/Th17 immune balance and the regulation of related cytokines.