葛根芩连汤对抗生素相关性腹泻模型肠道免疫及菌群的调节作用

Modulation of Intestinal Immunity and Bacterial Flora in an Antibiotic-Associated Diarrhoea Model by GeGen QinLian Decoction

关键词:  葛根芩连汤  抗生素相关性腹泻  肠道微生物  肠道免疫

Key words:  gegen qinlian decoction  antibiotic-associated diarrhoea  intestinal microorganisms  intestinal immunity

摘要:

目的 探讨葛根芩连汤通过调节肠道菌群以及提高肠道免疫功能来治疗抗生素相关性腹泻(antibiotic-associated diarrhoea,AAD)的作用机制。方法 将60只SD大鼠分为空白组、模型组、葛根芩连汤高、中、低剂量组、丽珠肠乐组,每组十只。除空白组外,其余组给予克林霉素诱导AAD模型后药物治疗2周。使用苏木精-伊红染色法(hematoxylin-eosin,HE)观察结肠形态学变化；免疫组织化学检测免疫相关因子活化诱导胞嘧啶脱氨酶(activation-induced cytidine deaminase,AICDA)、干扰素调节因子4(interferon regulatory factor 4,IRF4)、X-box结合蛋白1(x-box binding protein 1,XBP1)、BCL6、免疫球蛋白A(immunoglobulin A,IgA)的表达情况；宏基因组学测序方法分析各组大鼠粪便肠道菌群结构和功能变化。结果 葛根芩连汤和丽珠肠乐治疗后能修复AAD大鼠结肠形态、结构、黏膜上皮细胞损伤,上调免疫相关因子AICDA、BCL6、IRF4、XBP1、IgA的表达；宏基因组学结果显示,葛根芩连汤能改善肠道微生物多样性以及微生物丰度。在门水平上,厚壁菌门/拟杆菌门比值上升；在属水平上,Ligilactobacillus、Lactobacillus、Limosilactobacillus、Romboutsia、Corynebacterium、Enterococcus丰度上升；在肠道功能上,瓜氨酸、赖氨酸、半胱氨酸、鸟氨酸合成途径上调,肠道乳酸、乙酸、丙酸产生途径上调。结论 葛根芩连汤能改善 AAD大鼠炎症损伤,提高肠道免疫反应,调节肠道菌群结构和功能。

Abstract:

ABSTRACT: OBJECTIVE To investigate the mechanism of action of GeGen QinLian Decoction in treating AAD by regulating the intestinal flora as well as improving the intestinal immune function. METHODS Sixty SD rats were divided into a blank group, a model group, a high, medium and low dosage group of GeGen QinLian Decoction, and a group of Live Bifidobacterium Power, with ten rats in each group. Except for the blank group, the rest of the groups were given clindamycin to induce AAD model after drug treatment for 2 weeks. Morphological changes of the colon were observed using HE; immunohistochemistry was used to detect immunity-associated factor AICDA, IRF4, IRF4, XBP1, BCL6, and IgA; and macro-genomics sequencing was used to analyse the structural and functional changes in the faecal intestinal flora of rats in each group. RESULTS Treatment with GeGen QinLian Decoction and Live Bifidobacterium Power repaired the morphology, structure, and mucosal epithelial cell damage of the colon in AAD rats, and up-regulated the expression of immune-related factors AICDA, BCL6, IRF4, XBP1, and IgA. Macro-genomics results showed that GeGen QinLian Decoction improved the diversity of intestinal microorganisms as well as the microbial abundance. At the phylum level, the thick-walled bacterial phylum/anabolic bacterial phylum ratio increased; at the genus level, the abundance of Ligilactobacillus, Lactobacillus, Limosilactobacillus, Romboutsia, Corynebacterium, and Enterococcus increased; and at the intestinal function level. Citrulline, Lysine, Cysteine, Ornithine synthesis pathways were up-regulated, and intestinal lactic acid, acetic acid, propionic acid production pathways were up-regulated. CONCLUSION GeGen QinLian Decoction can improve inflammatory injury, enhance intestinal immune response, and regulate the structure and function of intestinal flora in AAD rats.