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痛泻要方通过调控肠道菌群-ENS-MM串扰减轻IBS-D结肠运动亢进的机制研究
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| 篇名： | 痛泻要方通过调控肠道菌群-ENS-MM串扰减轻IBS-D结肠运动亢进的机制研究 |
| TITLE： | Mechanism of Tongxie yaofang reducing colon hypermotility in IBS-D rats by regulating gut microbiota-ENS-MM crosstalk |
| 摘要： | 目的 探讨痛泻要方通过调控肠道菌群-肠神经系统（ENS）-肌层巨噬细胞（MM）串扰减轻腹泻型肠易激综合征（IBS-D）大鼠结肠运动亢进的作用机制。方法将40只新生雄性SD大鼠随机分为对照组、模型组、中药组[痛泻要方2.68g/（kg·d），以生药量计]、阳性对照组[双歧杆菌乳杆菌三联活菌片0.27g/（kg·d）]，每组10只。除对照组外，其余3组大鼠均采用母子分离+慢性束缚+番泻叶致泻法复制肝郁脾虚证型IBS-D模型。造模后，各药物组大鼠灌胃相应药液，每天1次，持续2周。于造模结束时和给药后观察各组大鼠的粪便性质（稀便出现率、稀便率）、结肠运动功能（小球排出时间）、内脏敏感性[不同压力下的腹部回缩反射（AWR）评分]；于给药后检测各组大鼠血清脂多糖（LPS）浓度，结肠组织中骨形态生成蛋白2（BMP2）、集落刺激因子1（CSF1）、Toll样受体4（TLR4）蛋白的表达和分布情况，以及肠道菌群的多样性、物种组成及差异。结果造模结束时，除对照组外的其余3组大鼠均100%出现稀便，稀便率和各压力下的AWR评分均较对照组显著升高，小球排出时间均较对照组显著缩短（P＜0.01或P＜0.05）。中药组和阳性对照组大鼠的稀便出现率分别为20%、80%；中药组大鼠稀便率、各压力下的AWR评分、血清LPS浓度和结肠组织肌层中CSF1、TLR4蛋白的表达均较模型组显著降低；小球排出时间，结肠组织肌层中BMP2蛋白的平均光密度，Chao1、Shannon、Faith’sPD、Simpson均较模型组显著延长或升高（P＜0.01或P＜0.05）；厚壁菌门/拟杆菌门相对丰度比值由低到高依次为中药组、对照组、阳性对照组、模型组，中药组大鼠的肠道菌群物种组成更贴近于对照组，其优势菌属包括普雷沃氏菌属、布劳特氏菌属等。结论痛泻要方能通过调节肠道菌群来调控肠道菌群-ENS-MM串扰关键蛋白BMP2、CSF1的表达，从而发挥减轻IBS-D大鼠结肠运动亢进的作用。 |
| ABSTRACT： | OBJECTIVE To study the effects of Tongxie yaofang reducing colon hypermotility in irritable bowel syndrome with diarrhea （IBS-D） rats by regulating gut microbiota-enteric nervous system （ENS）-muscularis macrophages （MM） crosstalk. METHODS Forty newborn male SD rats were randomly divided into control group， model group， TCM group [Tongxie yaofang 2.68 g/（kg·d）， calculated by raw material]， and positive control group [Live combined bifidobacterium and lactobacillus tablets 0.27 g/（kg·d）]， with 10 rats in each group. Except for the control group， the IBS-D model of liver stagnation and spleen deficiency syndrome was induced in the other 3 groups with the method of mother-child separation+chronic restraint+Folium Sennae- induced diarrhea. After modeling， the administration groups were given relevant drug liquid intragastrically， once a day， for consecutive 2 weeks. At the end of modeling and after administration， the fecal properties （the incidence and the rate of loose stools）， colonic motility （colon emptying time）， and visceral sensitivity [abdominal withdrawal reflex （AWR） scores under different pressures] of rats were observed in each group. The concentration of lipopolysaccharide （LPS） in serum was detected after medication， and the expressions and distribution of bone morphogenetic protein 2 （BMP2）， colony stimulating factor 1 （CSF1） and Toll-like receptor 4 （TLR4） in colon tissue were detected； the diversity， species composition and differences of gut microbiota were also determined. RESULTS At the end of modeling， compared with the control group， all rats of the other three groups suffered from loose stools （100%）， the rate of loose stools and AWR scores at different pressures increased significantly， and colon emptying time was shortened significantly （P＜0.01 or P＜0.05）. The incidences of loose stools were 20% in TCM group and 80% in the positive control group； the rate of loose stools and AWR scores at different pressures， serum concentration of LPS and protein expressions of CSF1 and TLR4 in muscle layer of colon tissue in TCM group significantly decreased， compared with the model group； colon emptying time， the average optical density of BMP2 protein in muscle layer of colon tissue， and the indexes of Chao 1， Shannon and Faith’s PD and Simpson E-mail：772699670@qq.com index of rats in TCM group were all prolonged or increased significantly， compared with the model group （P＜0.01 or P＜ E-mail：aiwangzi312@163.com 0.05）. The relative abundance ratio of Firmicutes/Bacteroidetes， from low to high， was in the order of TCM group， control group， positive control group and model group； the species composition of gut microbiota in TCM group was closer to control group， with dominant bacterial genera including Prevotella and Blautia. CONCLUSIONS Tongxie yaofang can regulate the expressions of BMP2 and CSF1， the key proteins of gut microbiota-ENS-MM crosstalk， by changing the gut microbiota， thus alleviating the abnormal hyperfunction of colon motility in IBS-D rats. |
| 期刊： | 2024年第35卷第13期 |
| 作者： | 彭龙；马佳乐；尹艺璇；步曦；赵双梅 |
| AUTHORS： | PENG Long，MA Jiale，YIN Yixuan，BU Xi，ZHAO Shuangmei |
| 关键字： | 痛泻要方；腹泻型肠易激综合征；肝郁脾虚证型；肠道菌群-ENS-MM串扰；骨形态发生蛋白2；集落刺激因子1 |
| KEYWORDS： | Tongxie yaofang； irritable bowel syndrome with diarrhea； liver stagnation and spleen deficiency syndrome； gut |
| 阅读数： | 3 次 |
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