Grasa Laura, Abecia Leticia, Forcén Raquel, Castro Marta, de Jalón José Antonio García, Latorre Eva, Alcalde Ana Isabel, Murillo María Divina
Department of Pharmacology and Physiology, Faculty of Veterinary Sciences, University of Zaragoza, Miguel Servet, 177, 50013, Zaragoza, Spain.
Department of Physiology and Biochemistry of Animal Nutrition, Estación Experimental del Zaidín (CSIC), Granada, Spain.
Microb Ecol. 2015 Oct;70(3):835-48. doi: 10.1007/s00248-015-0613-8. Epub 2015 Apr 21.
We examine the impact of changes in microbiota induced by antibiotics on intestinal motility, gut inflammatory response, and the function and expression of toll-like receptors (TLRs). Alterations in mice intestinal microbiota were induced by antibiotics and evaluated by q-PCR and DGGE analysis. Macroscopic and microscopic assessments of the intestine were performed in control and antibiotic-treated mice. TLR expression was determined in the intestine by q-RT-PCR. Fecal parameter measurements, intestinal transit, and muscle contractility studies were performed to evaluate alterations in intestinal motility. Antibiotics reduced the total bacterial quantity 1000-fold, and diversity was highly affected by treatment. Mice with microbiota depletion had less Peyer's patches, enlarged ceca, and mild gut inflammation. Treatment with antibiotics increased the expression of TLR4, TLR5, and TLR9 in the ileum and TLR3, TLR4, TLR6, TLR7, and TLR8 in the colon, and it reduced the expression of TLR2, TLR3, and TLR6 in the ileum and TLR2 and TLR9 in the colon. Antibiotics decreased fecal output, delayed the whole gut and colonic transit, and reduced the spontaneous contractions and the response to acetylcholine (ACh) in the ileum and colon. Activation of TLR4 by lipopolysaccharide (LPS) reverted the reduction of the spontaneous contractions induced by antibiotics in the ileum. Activation of TLR4 by LPS and TLR5 by flagellin reduced the response to ACh in the ileum in control mice. Our results confirm the role of the microbiota in the regulation of TLRs expression and shed light on the microbiota connection to motor intestinal alterations.
我们研究了抗生素诱导的微生物群变化对肠道运动、肠道炎症反应以及Toll样受体(TLR)功能和表达的影响。通过抗生素诱导小鼠肠道微生物群发生改变,并采用定量聚合酶链反应(q-PCR)和变性梯度凝胶电泳(DGGE)分析进行评估。对对照小鼠和经抗生素处理的小鼠进行肠道的宏观和微观评估。通过q-RT-PCR测定肠道中TLR的表达。进行粪便参数测量、肠道转运和肌肉收缩性研究,以评估肠道运动的改变。抗生素使细菌总量减少了1000倍,且多样性受到治疗的高度影响。微生物群耗竭的小鼠派尔集合淋巴结较少、盲肠增大且有轻度肠道炎症。抗生素治疗增加了回肠中TLR4、TLR5和TLR9以及结肠中TLR3、TLR4、TLR6、TLR7和TLR8的表达,同时降低了回肠中TLR2、TLR3和TLR6以及结肠中TLR2和TLR9的表达。抗生素减少了粪便排出量,延迟了全肠道和结肠的转运,并降低了回肠和结肠的自发收缩以及对乙酰胆碱(ACh)的反应。脂多糖(LPS)激活TLR4可逆转抗生素诱导的回肠自发收缩减少。LPS激活TLR4以及鞭毛蛋白激活TLR5可降低对照小鼠回肠对ACh的反应。我们的结果证实了微生物群在调节TLR表达中的作用,并揭示了微生物群与肠道运动改变之间的联系。