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双歧杆菌和乳酸杆菌促进便秘小鼠肠嗜铬细胞 5-羟色胺分泌的机制。

Mechanisms underlying the promotion of 5-hydroxytryptamine secretion in enterochromaffin cells of constipation mice by Bifidobacterium and Lactobacillus.

机构信息

College of Food Science and Engineering, Ocean University of China, Qingdao, China.

Qingdao Central Hospital, Qingdao, China.

出版信息

Neurogastroenterol Motil. 2021 Jul;33(7):e14082. doi: 10.1111/nmo.14082. Epub 2021 Jan 15.

Abstract

BACKGROUND

5-Hydroxytryptamine (5-HT) could play a role in alleviating symptoms in constipation. However, the mechanism underlying the role of intestinal flora in the promotion of 5-HT secretion by enterochromaffin cells (ECs) and regulation of the gastrointestinal endocrine system remains unclear.

METHODS

A constipation mouse model was constructed, and the 5-HT, chromogranin A (CGA), substance P (SP), motilin (MTL), dopamine, and noradrenaline expression levels were measured using enzyme-linked immunosorbent assay(Elisa) and immunofluorescence, and key proteins, such as the transient receptor potential (TRP) ion channels/tryptophan hydroxylase (TPH) and olfactory receptor (OR), were determined using western blot. Flow cytometry and in vivo imaging were used to observe microbial colonization in the intestinal tracts of mice.

KEY RESULTS

Bifidobacterium animalis F1-7 (F1-7), Lactobacillus paraccasei F34-3 (F34-3), and Lactobacillus plantarum FWDG (FWDG) promoted 5-HT secretion. F1-7 and F34-3 induced CGA expression, increased catecholamine secretion, and activated the CGA/α2A adrenoreceptor (ADRα2A) cascade signal in ECs. FWDG increased noradrenaline levels and activated the ADRα2A signal in ECs. SP content increased in F1-7 and F34-3, and MTL expression increased in FWDG via the above signal. F1-7 and F34-3 downregulated TRPV4 and upregulated TPH, whereas FWDG upregulated OR2A4 for promoting 5-HT secretion by ECs. Finally, we observed that F1-7, F34-3, and FWDG were well colonized in the large intestine.

CONCLUSIONS AND INFERENCES

F1-7, F34-3, and FWDG promoted 5-HT secretion in ECs of constipation mice by activating the CGA/ADRα2A cascade signal and regulating the TRP/TPH-OR pathways.

摘要

背景

5-羟色胺(5-HT)可能在缓解便秘症状方面发挥作用。然而,肠道菌群促进肠嗜铬细胞(ECs)分泌 5-HT 并调节胃肠内分泌系统的机制尚不清楚。

方法

构建便秘小鼠模型,采用酶联免疫吸附法(ELISA)和免疫荧光法检测 5-HT、嗜铬粒蛋白 A(CGA)、P 物质(SP)、胃动素(MTL)、多巴胺和去甲肾上腺素的表达水平,采用蛋白质印迹法检测瞬时受体电位(TRP)离子通道/色氨酸羟化酶(TPH)和嗅觉受体(OR)等关键蛋白。采用流式细胞术和体内成像观察小鼠肠道微生物定植情况。

主要结果

动物双歧杆菌 F1-7(F1-7)、副干酪乳杆菌 F34-3(F34-3)和植物乳杆菌 FWDG(FWDG)促进 5-HT 分泌。F1-7 和 F34-3 诱导 CGA 表达,增加儿茶酚胺分泌,激活 ECs 中的 CGA/α2A 肾上腺素能受体(ADRα2A)级联信号。FWDG 增加去甲肾上腺素水平并激活 ECs 中的 ADRα2A 信号。F1-7 和 F34-3 增加 SP 含量,FWDG 通过上述信号增加 MTL 表达。F1-7 和 F34-3 下调 TRPV4 并上调 TPH,而 FWDG 上调 OR2A4 以促进 ECs 分泌 5-HT。最后,我们观察到 F1-7、F34-3 和 FWDG 在大肠中定植良好。

结论和推论

F1-7、F34-3 和 FWDG 通过激活 CGA/ADRα2A 级联信号和调节 TRP/TPH-OR 途径促进便秘小鼠 ECs 中 5-HT 的分泌。

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