柴参解郁汤对腹泻型肠易激综合征小鼠粪便胆汁酸代谢轮廓的影响
Effects of Chaishen Jieyu Decoction on the Metabolic Profile of Fecal Bile Acids in Mice with Diarrhea Predominant Irritable Bowel Syndrome
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摘要:目的 通过研究柴参解郁汤对腹泻为主型肠易激综合征(Diarrhea predominant irritable bowel syndrome, IBS-D)模型小鼠粪便中胆汁酸代谢轮廓的影响, 探讨柴参解郁汤改善IBS-D的可能机制。方法 将C57BL/6小鼠随机分为3组, 空白组、模型组、柴参解郁汤组(15 g·kg-1)。IBS-D小鼠模型采用束缚性应激复合灌胃番泻叶造模, 造模成功后, 灌胃给予柴参解郁汤2周, 分析小鼠粪便含水量, 检测空肠病理损伤, Western blot检测空肠中COX-2、iNOS的蛋白表达, 通过检测FITC标记的右旋糖苷及紧密连接蛋白ZO-1、OCCLUDIN的mRNA表达，测定小鼠胃肠道通透性。进一步采用靶向代谢组学分析小鼠粪便胆汁酸代谢轮廓。结果 与空白组相比, 模型组小鼠粪便含水量增高(P < 0.01), 给予柴参解郁汤后粪便含水量下降(P < 0.01);与空白组相比, 模型组小鼠空肠中COX-2和iNOS蛋白表达量上调(P < 0.05，P < 0.01), 而柴参解郁汤可下调空肠细胞中iNOS蛋白表达(P < 0.01);与空白组相比，模型组小鼠空肠中ZO-1、OCCLUDIN的mRNA表达量明显下降(P < 0.01)，FIFC-右旋糖苷含量下降(P < 0.01)，柴参解郁汤治疗后表达量有所回调(P < 0.05，P < 0.01)。与空白组相比, 模型组小鼠空肠绒毛大部分脱落坏死, 绒毛的固有层细胞溶解, 绒毛结构模糊不清, 给予柴参解郁汤后趋于正常。IBS-D小鼠粪便中胆汁酸组成和含量发生显著改变, 柴参解郁汤可显著回调模型组小鼠的胆汁酸代谢紊乱, 如石胆酸(LCA)、去氧胆酸(DCA)、鹅去氧胆酸(CDCA)等。结论 柴参解郁汤可有效缓解IBS-D小鼠空肠组织病理改变并能改善IBS-D小鼠胆汁酸异常变化, 这可能是由于柴参解郁汤修复肝肠循环胆汁酸合成和转化途径, 增加结合型胆汁酸含量而起到保护肠道的作用。Abstract:OBJECTIVE To investigate therapeutic effect and potential mechanism of the Chaishen Jieyu Decoction on IBS-D mice model from regulating the fecal bile acid profile.METHODS The C57BL/6 mice were randomly divided to 3 groups, normal mice, IBS-D mice and the Chaishen Jieyu Decoction intervention group. IBS-D mice were induced by the resisting immobilization stress and gavage of Folium senna. We studied the pathological changes of the mouse jejunum sections by HE, the protein expression of COX-2 and iNOS in jejunum, FITC-dextran and detected the relative mRNA expressions of main tight junctions proteins(ZO-1 & OCCLUDIN) in mice to determine the gastrointestinal permeability and analysis of fecal bile acid profile in mice using targeted metabolomics method.RESULTS Compared with the control group, the fecal water content had an uptrend in the model group, showed a decreasing trend after treatment with Chaishen Jieyu Decoction. COX-2 and iNOS protein expression was upregulated in the jejunum of mice in the model group when compared to the control group, and Chaishen Jieyu Decoction could downregulate iNOS protein expression in jejunum (P < 0.01), and the mouse jejunum mRNA expression of ZO-1 and OCCLUDIN in model group were significantly decreased when compared with the control group(P < 0.01), while CSJY Decoction could increase these two proteins' mRNA expression(P < 0.05, P << 0.01);Also, compared with the control group, the pathological alterations such as incomplete jejunal structural organization, disorganized intestinal epithelial cell arrangement, crypt damage as well as intestinal epithelial integrity in the model group mice tended to be normalized after administration of CSJY Decoction. the composition and content of fecal bile acids in IBS-D mice were altered, after Chaishen Jieyu Decoction intervention, the disturbed fecal bile acid metabolism and disposition were significantly altered, especially for lithic cholic acid (LCA), deoxycholic acid (DCA) and deoxycholic acid (CDCA).CONCLUSION Chaishen Jieyu Decoction has therapeutic effect on IBS-D mice and this effect may be partially contributed to regulating the bile acids metabolism.
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