探讨莫沙必利改善心理应激小鼠5-羟色胺代谢通路、肠道菌群以及食管炎症的作用机制。

选择SPF级雄性8周龄的昆明小鼠数字化随机分为正常对照组、心理应激组及莫沙必利干预组（每组6只）；心理应激组和莫沙必利组小鼠每天在自制式束缚器中限制活动2 h，其余时间3组小鼠在相同环境中自由饮水摄食，实验持续14 d；通过HE染色观察食管病理学改变；采用RT-PCR方法检测分析3组食管组织中色氨酸（TPH）、色氨酸羟化酶-1（TPH-1）、吲哚胺2，3加双氧酶-1（IDO-1）、色氨酸2，3-双加氧酶（TDO）、犬尿酸（KYN）、5-羟色胺（5-HT）、5-HT4受体（5-HT4R）、单核细胞/巨噬细胞表面标志物（F4/80、CD68）及炎症因子[白细胞介素-1β（IL-1β）IL-1β、Toll样受体4(TLR-4)、肿瘤坏死因子-α（TNF-α）]的mRNA表达水平；利用16s rRNA高通量测序技术检测3组小鼠肠道菌群，并对比分析3组小鼠的菌群组成差异。

心理应激组显著降低5-HT及5-HT4R的mRNA表达水平（P<0.001）；而莫沙必利干预显著增高5-HT及5-HT4R的mRNA水平（P<0.01）。心理应激组显著升高食管组织中5-HT代谢通路中的酶物质（IDO-1、TDO、KYN）的mRNA水平（P<0.01、P<0.001）；同时其显著降低TPH-1和TPH的mRNA表达水平（P<0.01、P<0.001）；莫沙必利干预能够改善心理应激诱导上述5-HT代谢通路酶物质的异常表达（P<0.01）。心理应激组小鼠菌群多样性较低；莫沙必利干预显著改善心理应激小鼠Helicobacteraceae（螺杆菌科）、Alloprevotella（拟普雷沃菌属）、Lactobacillaceae（乳杆菌科）等菌群的异常变化。莫沙必利显著抑制心理应激诱导食管粘膜炎症浸润反应、免疫细胞（CD68、F4/80）以及炎症因子（IL-1β、TLR-4、TNF-ɑ）的高表达（P<0.001）。

莫沙必利通过调节5-羟色胺代谢途径抑制心理应激诱发肠道菌群结构异常及食管炎症的发生。

To explore the mechanism of Mosapride in improving 5-hyd roxytryptamine metabolic pathway, gut bacteria and esophageal inflammation in mice under stress.

8 week old SPF male mice were randomly divided into control, stress and mosapride intervention group (n=6); The mice in the stress and mosapride were restricted in the self-made restraint device for 2h every day and kept for 14 days period; The pathological changes of esophagus were observed by HE staining; tryptophan (TPH), tryptophan hydroxylase-1 (TPH-1), indoleamine 2,3 dioxygenase 1 (IDO-1), tryptophan 2,3 dioxygenase (TDO), canine uric acid (KYN), 5-HT,5-HT4R, F4/80, CD68 and inflammatory factors [interleukin-1 β (IL-1 β), toll-like receptor-4 (TLR-4), tumor necrosis factor-α (TNF-ɑ)] mRNA level were detected by RT-PCR; The 16s rRNA high-throughput sequencing technology was used to detect the intestinal flora of mice.

The mRNA expression levels of 5-HT and 5-HT4R were decreased in the stress group (P<0.001); Mosapride increased the mRNA levels of 5-HT and 5-HT4R (P<0.01). The mRNA level of enzymes (IDO-1, TDO, KYN) in 5-HT metabolic pathway was increased in the stress group (P<0.01, P<0.001); stress reduced the mRNA expression levels of TPH-1 and TPH (P<0.01, P<0.001); Mosapride intervention can improve the abnormal expression of 5-HT metabolic pathway enzymes induced by stress (P<0.01). The microbial diversity of mice in stress group was low; Mosapride intervention significantly improved the abnormal changes of Helicobacterceae, Alloprevotella and Lactobacillus in stress mice. Mosapride significantly inhibited the infiltration of esophageal mucositis induced by stress, and suppressed over expressions of immune cells (CD68, F4/80) and inflammatory factors(IL-1β, TLR-4, TNF-ɑ) (P<0.001).

Mosapride can inhibit the occurrence of abnormal intestinal flora structure and esophageal inflammation induced by stress through regulating 5-HT metabolism.