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半夏泻心汤通过调节肠道微生物群及其代谢物缓解 DSS 诱导的溃疡性结肠炎。

Banxia Xiexin decoction modulates gut microbiota and gut microbiota metabolism to alleviate DSS-induced ulcerative colitis.

机构信息

Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.

Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China; Institute of Traditional Chinese Medicine Health Industry, China Academy of Chinese Medical Sciences, Nanchang, 330000, China; Jiangxi Health Industry Institute of Traditional Chinese Medicine, Nanchang, 330000, China.

出版信息

J Ethnopharmacol. 2024 May 23;326:117990. doi: 10.1016/j.jep.2024.117990. Epub 2024 Feb 27.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Banxia Xiexin decoction (BXD) is a classic traditional Chinese medicine prescription for treating ulcerative colitis (UC). However, its potential mechanism of action is still unclear.

AIM OF THE STUDY

Reveal the correlation between the beneficial impacts of BXD on UC and the composition of the gut microbiota.

MATERIALS AND METHODS

The major constituents of BXD were identified using the HPLC-DAD technique. An experimental model of UC was induced in male C57BL/6 mice by administering dextran sodium sulfate (DSS). A total of 48 mice were divided into different groups, including control, model, high-dose BXD treatment, medium-dose BXD treatment, low-dose BXD treatment, and a group treated with 5-amino acid salicylic acid (5-ASA). Body weight changes and disease activity index (DAI) scores were documented; colon length, colon index, spleen index, and thymus index scores were determined; myeloperoxidase (MPO) and tumor necrosis factor-α (TNF-α) activities were assessed; and histological staining with hematoxylin-eosin and alcian blue/phosphate Schiff was performed. The immunofluorescence technique was employed to examine the presence of ZO-1 and occludin in the colon tissue. 16S rRNA sequencing was employed to assess the gut microbiota's diversity and metabolomics was utilized to examine alterations in metabolites within the gut microbiota. The impact of BXD on the gut microbiota was confirmed through fecal microbiota transplantation (FMT).

RESULTS

BXD exhibited a positive impact on UC mice, particularly in the high-dose BXD treatment group. The BXD group experienced weight recovery, decreased DAI scores, improved colon length, and restored of spleen and thymus index scores compared to the DSS group. Additionally, BXD alleviated colon damage and the inflammatory response while restoring intestinal barrier function. FMT in BXD-treated mice also showed therapeutic effects in UC mice. At the phylum level, the relative abundance of Desulfobacterota, Deferribacterota and Actinobacteriota increased; at the genus level, g__norank__f__Muribaculaceae, Dubosiella, Akkermansia, and Lactobacillus increased, whereas Faecalibaculum, Alloprevotella, Turicibacter, and g_Paraprevotella decreased. g__norank_f__Muribaculaceae was positively correlated with body weight and colon length and negatively with colon index scores, splenic index scores, and MPO levels; Alloprevotella was positively correlated with splenic index scores, histological scores, and TNF-α levels and negatively with thymus index scores and thymus index scores. Faecalibaculum was positively correlated with colon index scores and MPO levels. Metabolic investigations revealed 58 potential indicators, primarily associated with the metabolism of amino acids, purines, and lipids. Alloprevotella, g_Paraprevotella, and Bifidobacterium were strongly associated with metabolic pathways.

CONCLUSION

BXD showed beneficial therapeutic effects in UC mice. The mechanism may be by promoting the balance and variety of gut microbiota, as well as regulating the metabolism of amino acids, purines, and lipids.

摘要

民族药理学相关性

半夏泻心汤(BXD)是治疗溃疡性结肠炎(UC)的经典中药方剂。然而,其潜在的作用机制尚不清楚。

研究目的

揭示 BXD 对 UC 的有益影响与肠道微生物群组成之间的相关性。

材料和方法

采用高效液相色谱-二极管阵列检测(HPLC-DAD)技术鉴定 BXD 的主要成分。通过给予葡聚糖硫酸钠(DSS)诱导雄性 C57BL/6 小鼠建立 UC 实验模型。将 48 只小鼠分为对照组、模型组、BXD 高剂量治疗组、BXD 中剂量治疗组、BXD 低剂量治疗组和 5-氨基水杨酸(5-ASA)治疗组。记录体重变化和疾病活动指数(DAI)评分;测定结肠长度、结肠指数、脾指数和胸腺指数评分;评估髓过氧化物酶(MPO)和肿瘤坏死因子-α(TNF-α)活性;并进行苏木精-伊红和阿尔辛蓝/磷钨酸染色的组织学染色。采用免疫荧光技术检测结肠组织中 ZO-1 和闭合蛋白的存在。采用 16S rRNA 测序评估肠道微生物群的多样性,采用代谢组学检测肠道微生物群中代谢物的变化。通过粪便微生物群移植(FMT)证实 BXD 对肠道微生物群的影响。

结果

BXD 对 UC 小鼠有积极影响,尤其是在 BXD 高剂量治疗组。BXD 组与 DSS 组相比,体重恢复、DAI 评分降低、结肠长度增加、脾指数和胸腺指数评分恢复。此外,BXD 减轻了结肠损伤和炎症反应,同时恢复了肠道屏障功能。BXD 治疗小鼠的 FMT 也显示出对 UC 小鼠的治疗效果。在门水平上,脱硫杆菌门、脱硫菌门和放线菌门的相对丰度增加;在属水平上,g__norank__f__Muribaculaceae、Dubosiella、Akkermansia 和 Lactobacillus 增加,而 Faecalibaculum、Alloprevotella、Turicibacter 和 g_Paraprevotella 减少。g__norank_f__Muribaculaceae 与体重和结肠长度呈正相关,与结肠指数评分、脾指数评分和 MPO 水平呈负相关;Alloprevotella 与脾指数评分、组织学评分和 TNF-α水平呈正相关,与胸腺指数评分和胸腺指数评分呈负相关。Faecalibaculum 与结肠指数评分和 MPO 水平呈正相关。代谢研究发现了 58 个潜在指标,主要与氨基酸、嘌呤和脂质的代谢有关。Alloprevotella、g_Paraprevotella 和双歧杆菌与代谢途径密切相关。

结论

BXD 对 UC 小鼠有有益的治疗作用。其机制可能是通过促进肠道微生物群的平衡和多样性,以及调节氨基酸、嘌呤和脂质的代谢。

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