基于16S rRNA技术分析桔梗超微粉对小鼠肠道菌群多样性的影响

doi:10.16431/j.cnki.1671-7236.2025.07.036

摘要/Abstract

摘要： 【目的】观察桔梗超微粉对小鼠肠道菌群多样性的影响，为桔梗超微粉的临床应用及其安全性提供依据。【方法】将24只雄性昆明小鼠随机分为对照组、桔梗超微粉高(3.0 g/kg)、中(1.5 g/kg)、低(0.3 g/kg)剂量组，每组6只。桔梗超微粉各剂量组小鼠灌胃相应剂量溶于水后的桔梗超微粉，对照组灌胃等体积纯化水，连续给药14 d，每日记录体重和耗料量。在试验第15天处死各组小鼠进行剖检及组织病理学观察，同时无菌取小鼠粪便，扩增粪便菌群16S rRNA基因的V3-V4区，采用高通量测序分析菌群Alpha和Beta多样性及门和属水平的物种组成并进行微生物功能预测分析。【结果】 ①桔梗超微粉中、高剂量组小鼠平均体增重均显著降低于对照组(P＜0.05)，桔梗超微粉中剂量组小鼠平均耗料量明显低于对照组。②与对照组相比，桔梗超微粉各剂量组小鼠胃、十二指肠、空肠和回肠组织均未见病理学异常。③桔梗超微粉各剂量组小鼠肠道菌群Alpha多样性指数均高于对照组(P＞0.05)，且随着桔梗超微粉使用剂量增多呈上升趋势。肠道菌群Beta多样性分析结果显示，与对照组相比，低、高剂量桔梗超微粉对小鼠肠道菌群结构影响不显著(P>0.05)，中剂量桔梗超微粉可显著改变小鼠肠道菌群结构(P＜0.05)。④与对照组相比，在门水平上，桔梗超微粉中剂量组小鼠肠道内厚壁菌门和蓝藻门丰度显著增加(P＜0.05)，拟杆菌门、变形菌门和浮霉菌门丰度显著下降(P＜0.05)；在属水平上，中剂量组小鼠肠道内普雷沃菌属、副拟杆菌属和rc4_4丰度显著增加(P＜0.05)，Kaistobacter属和假单胞菌属等丰度显著下降(P＜0.05)。⑤微生物功能预测分析发现，桔梗超微粉中剂量组小鼠肠道微生物基因组功能均主要富集在氨基酸生物合成及辅因子、假体基团、电子载体与维生素生物合成和核苷酸生物合成等方面。与对照组相比，桔梗超微粉中剂量组存在11条显著下调的差异代谢通路，主要涉及有氟苯甲酸酯降解、多环芳烃降解和酵母减数分裂等。【结论】桔梗超微粉可增加小鼠肠道菌群多样性，其中1.5 g/kg桔梗超微粉可显著改变肠道菌群结构。本试验结果为桔梗超微粉临床使用的科学用量及安全性提供数据支持，同时为今后桔梗超微粉与肠道菌群相关的药效学研究提供了数据支持。

关键词: 桔梗超微粉; 16S rRNA; 小鼠; 肠道菌群; 微生物多样性

Abstract: 【Objective】 The purpose of this study was to observe the effect of Platycodon grandiflorum ultrafine powder on the diversity of intestinal flora in mice,and provide evidence for clinical application and safety of Platycodon grandiflorum ultrafine powder. 【Method】 Twenty-four male Kunming mice were randomly divided into control group and high (3.0 g/kg),medium (1.5 g/kg) and low (0.3 g/kg) dose groups of Platycodon grandiflorum ultrafine powder，6 mice in each group.Mice in each dose groups of Platycodon grandiflorum ultrafine powder were gavaged with the corresponding dose of Platycodon grandiflorum ultrafine powder dissolved in water,and mice in control group were gavaged with equal volume of purified water,the medication was administered continuously for 14 days.The body weight and feed consumption of mice in each group were recorded daily.On the 15th day of the experiment,mice in each group were sacrificed for autopsy and histopathological observation.Meanwhile,the feces of mice were aseptically collected to amplify the V3-V4 regions of the 16S rRNA gene of the fecal microbiota.High-throughput sequencing was used to analyze Alpha and Beta diversity of the microbiota and the species composition at phylum and genus levels.And microbial function prediction and analysis were conducted. 【Result】 ①The body weight gain of mice in medium and high dose groups were significantly lower than those in control group (P＜0.05).The average feed consumption of mice in medium dose group of Platycodon grandiflorum superfine powder was lower than that in control group.②Compared with control group,no pathological abnormalities were found in stomach,duodenum,jejunum and ileum of mice in each dose group of Platycodon grandiflorum superfine powder.③The Alpha diversity index in intestinal flora of mice in each dose group of Platycodon grandiflorum superfine powder was higher than that in control group(P＞0.05),and increased with the increase of the dosage.Beta diversity analysis of intestinal flora showed that low and high doses of Platycodon grandiflorum superfine powder had no significant effect on the structure of intestinal flora in mice,while medium doses of Platycodon grandiflorum superfine powder could significantly change the structure of intestinal flora in mice (P＜0.05).④Compared with control group,at phylum level,the contents of Firmicutes and Cyanobacteria in the intestinal tract of mice in medium dose group of Platycodon grandiflorum ultrafine powder were significantly increased (P＜0.05),while the abundance of Bacteroidetes,Proteobacteria and Planctomycetes was significantly decreased (P＜0.05).At genus level,the abundance of Prevotella,Parabacteroides and rc4_4 in the intestinal tract of mice in medium dose group was significantly increased (P＜0.05),while the abundance of Kaistobacter and Pseudomonas etc.was significantly decreased (P＜0.05).⑤Microbial function prediction analysis found that the intestinal microbial genome functions of mice in medium dose group of Platycodon grandiflorum superfine powder were mainly enriched in amino acid biosynthesis and cofactors,prosthetic groups,electronic carriers and vitamin biosynthesis and nucleotide biosynthesis.Compared with control group,there were 11 significantly downregulated differential metabolic pathways in medium dose group of Platycodon grandiflorum superfine powder,mainly involving the degradation of fluorobenzoates,polycyclic aromatic hydrocarbons and yeast meiosis. 【Conclusion】 Platycodon grandiflorum ultrafine powder was able to increase the diversity of intestinal flora in mice,and 1.5 g/kg Platycodon grandiflorum ultrafine powder could significantly change the structure of intestinal flora.The results of this experiment provided data support for the scientific dosage and safety of Platycodon grandiflorum ultrafine powder’s clinical use,and also support for future pharmacodynamic studies related to Platycodon grandiflorum ultrafine powder and intestinal flora.

Key words: Platycodon grandiflorum ultrafine powder; 16S rRNA; mice; intestinal flora; microbial diversity

中图分类号:

S853.74

李芷珊, 周照月, 刘天龙, 王帅玉, 江海洋. 基于16S rRNA技术分析桔梗超微粉对小鼠肠道菌群多样性的影响[J]. 中国畜牧兽医, 2025, 52(7): 3372-3385.

LI Zhishan, ZHOU Zhaoyue, LIU Tianlong, WANG Shuaiyu, JIANG Haiyang. Analysis of the Effects of Platycodon grandiflorum Ultrafine Powder on the Diversity of Intestinal Flora in Mice Based on 16S rRNA Technology[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3372-3385.

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