痛泻安肠方对肠易激综合征小鼠短链脂肪酸代谢及Th1/Th2细胞免疫平衡的影响

The Effect of Tongxie Anchang Decoction on Short-chain Fatty Acids Metabolism and Th1/Th2 Cell Immune Balance in Mice with Irritable Bowel Syndrome

目的观察痛泻安肠方对肝郁脾虚证腹泻型肠易激综合征(IBS-D)小鼠模型短链脂肪酸代谢及Th1/Th2细胞免疫平衡的影响。方法30只18~22 g的雄性C57BL/6小鼠随机分为空白组、模型组、痛泻安肠方低剂量组、痛泻安肠方中剂量组、痛泻安肠方高剂量组,每组6只。采用束缚应激联合番泻叶灌胃复制IBS-D小鼠模型,造模成功后分别予相应浓度中药灌胃14 d,每日1次,通过靶向代谢组学检测小鼠粪便中短链脂肪酸含量的变化,并通过流式细胞术检测肠系膜淋巴结中CD3+CD4+INF-γ+T细胞(Th1细胞)、CD3+CD4+IL-4+T细胞(Th2细胞)的占比情况。结果与空白组相比,模型组小鼠粪便短链脂肪酸(SCFAs)相对含量显著升高(P<0.01),尤其是乙酸、丙酸及丁酸的含量均显著升高(P<0.01或P<0.05);不同剂量痛泻安肠方治疗后,粪便总SCFAs相对含量显著降低(P<0.01),乙酸、丙酸及丁酸均有不同程度降低,其中痛泻安肠方各剂量组均可显著降低乙酸和丁酸的含量,差异具有统计学意义(P<0.01),低剂量组及中剂量组可显著降低丙酸的含量,差异具有统计学意义(P<0.01)。造模后,肠易激综合征小鼠肠系膜淋巴结的Th2细胞占比明显降低,而Th1细胞占比及Th1/Th2细胞比率明显升高,差异具有统计学意义(P<0.05);治疗后,痛泻安肠方各剂量组Th2细胞占比均有升高,Th1细胞占比及Th1/Th2细胞比率降低。结论痛泻安肠方可以降低IBS-D小鼠粪便SCFAs的含量,调节小鼠Th1/Th2细胞免疫紊乱。

Objective:To investigate the modulating effect of Tongxie Anchang Decoction on short-chain fatty acids metabolism and Th1/Th2 cell immune balance in mice of irritable bowel syndrome with predominant diarrhea(IBS-D)with syndrome of stagnation of liver qi and spleen deficiency.Methods:30 male C57BL/6 mice,weigh⁃ing 18~22 g,were randomly divided into control group,model group,Tongxie Anchang Decoction low group,Tongx⁃ie Anchang Decoction medium group,and Tongxie Anchang Decoction high group,with 6 mice in each group.The IBS-D mouse model was replicated using restraint stress combined with senna leaf gavage.After successful model⁃ing,corresponding dose of Tongxie Anchang Decoction were administered by gavage once a day for 14 days.The changes in short chain-fatty acid content in the feces of mice were detected by targeted metabolomics.The propor⁃tion of CD3+CD4+INF-γ+T cells(Th1 cells)and CD3+CD4+IL-4+T cells(Th2 cells)in mesenteric lymph nodes were detected by flow cytometry.Results:Compared with the control group,the relative content of fecal shortchain fatty acids(SCFAs)of mice in the model group increased(P<0.01),with butyric acid,propionic acid and acetic acid all significantly increased(P<0.01 or P<0.05).After treatment with the different dose of Tongxie An⁃chang Decoction,the relative content of total SCFAs in feces decreased(P<0.01),while butyric acid,propionic ac⁃id and acetic acid all decreased to varying degrees.Among them,each dose group of Tongxie Anchang Decoction significantly reduced the content of acetic acid and butyric acid(P<0.01).The low dose and medium dose groups significantly reduced the content of propionic acid(P<0.01).In the model group,the proportion of Th2 cells in mouse mesenteric lymph nodes significantly decreased,while the proportion of Th1 cells and Th1/Th2 significantly increased,with a statistically significant difference(P<0.05);after treatment,the proportion of Th2 cells in each dose group of Tongxie Anchang Decoction increased,while the proportion of Th1 cells and Th1/Th2 decreased.Conclusion:Tongxie Anchang Decoction can reduce the content of SCFAs in the feces of IBS-D mice with syn⁃drome of stagnation of liver qi and spleen deficiency,and regulate Th1/Th2 cell immune disorders in mice.