双参平肺方对博来霉素致肺纤维化大鼠肠道菌群的影响

陈叶青; 孟小艺; 范欣生

doi:10.14148/j.issn.1672-0482.2023.0257

双参平肺方对博来霉素致肺纤维化大鼠肠道菌群的影响

doi: 10.14148/j.issn.1672-0482.2023.0257

南京中医药大学中医学院·中西医结合学院, 江苏南京 210023

基金项目:

国家自然科学基金青年科学基金项目 81904084

国家自然科学基金面上项目 82174258

南京中医药大学自然科学基金青年项目 NZY81904084

江苏省中药资源产业化过程协同创新中心重点项目 ZDXM-2020-22

详细信息

作者简介:
陈叶青, 女, 讲师, E-mail: chenyeqing@njucm.edu.cn

通讯作者:
范欣生, 女, 研究员, 主要从事经典方剂方证药理效应及机制研究, E-mail: shshfan@njucm.edu.cn

中图分类号: R285.5
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- 被引次数: 0
出版历程
- 收稿日期: 2022-10-05
- 网络出版日期: 2023-03-21
- 发布日期: 2023-03-10

Effect of Shuangshen Pingfei Formula on Gut Microbiota in BLM-Induced Pulmonary Fibrosis Rats

College of Chinese Medicine, College of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China

摘要

摘要: 目的研究博来霉素(BLM)致肺纤维化(PF)及给予双参平肺方对肠道菌群结构的影响。方法采用气管内滴注BLM(5 mg·kg^-1)复制PF大鼠模型, 将造模后的大鼠随机分为模型组、双参平肺方组, 并设空白对照组, 每组6~8只, 双参平肺方组灌胃双参平肺方6.5 g·kg^-1, 模型组、空白对照组灌胃等量生理盐水, 连续灌胃28 d后进行肺组织病理学评价, 并测定肺组织羟脯氨酸(HYP)含量及金属酶蛋白7(MMP7)mRNA水平, 采用16S rDNA测序技术检测肠道菌群变化, 通过相关性分析, 筛选与双参平肺方改善PF程度相关的菌群, 通过ROC分析评估肠道菌群识别PF及双参平肺方对其作用的可能性。结果与空白对照组比较, 模型组大鼠肺组织出现病变, HYP含量及MMP7 mRNA水平显著升高(P < 0.01);给予双参平肺方后, 上述指标较模型组均显著改善(P < 0.01)。16S rDNA测序结果显示, 模型组有17个肠道菌发生了改变, 给予双参平肺方后能改善这一变化, 结合相关性分析, 发现双参平肺方能下调部分致病菌属如梭杆菌属、志贺氏菌属、异杆菌属、梭菌属等的丰度, 同时上调部分有益菌属如凝聚杆菌属、柯林斯氏菌属、双歧杆菌属、乳杆菌属等菌属的丰度。ROC分析表明使用梭杆菌鉴定PF及验证双参平肺方改善作用是可靠的。结论 PF导致肠道菌群紊乱, 给予双参平肺方后可通过上调益生菌并下调致病菌的丰度改善这一紊乱。
- 肺纤维化 /
- 双参平肺方 /
- 肠道菌群 /
- 16S rDNA /
- ROC分析 /
- 梭杆菌
Abstract: OBJECTIVE To explore the effect of Shuangshen Pingfei formula (SSPF) on gut microbiota in bleomycin (BLM)-induced pulmonary fibrosis rats. METHODS The PF rat model was established by intratracheal instillation of BLM. The rats were randomly divided into three groups (n=6-8): saline group, BLM+saline group, BLM+SSPF group. SSPF (6.5 g·kg^-1) was given every day via gavage starting after 1 days of BLM/normal saline instillation. Equal volumes of saline were given to rats in the other two groups. At 28 days post-treatment, rat lungs were harvested to detect the lung coefficient and the status of lung inflammation and fibrosis, and hydroxyproline (HYP) content and metalloprotein 7 (MMP7) mRNA expression were measured. 16S rDNA sequencing technology was applied to detect intestinal microflora changes. The microbiota related to SSPF-induced improvement of fibrosis was screened out by correlation analysis. The possibility of gut microbiota in identifying pulmonary fibrosis and SSPF-induced anti-pulmonary fibrosis activity was assessed by ROC analysis. RESULTS Histological analysis showed that chronic inflammation and pulmonary fibrosis occurred 28 days after BLM instillation, and decreased after SSPF administration. BLM increased HYP content and MMP7 mRNA expression, which were reversed by SSPF treatment. 17 genera of microflora were altered in the model, which were improved after administration of SSPF. Combined with correlation analysis, SSPF decreased the abundance of some pathogenic bacteria such as Fusobacterium, Shigella, Allobaculum and Clostridium, and increased the abundance of some beneficial bacteria such as Aggregatibacter, Collinsella, Bifidobacterium and Lactobacillus. ROC analysis indicated that it was dependable to identify pulmonary fibrosis and verify SSPF-induced improvement in pulmonary fibrosis by using gut microorganisms. CONCLUSION Pulmonary fibrosis can cause the changes in gut microbiota, and SSPF can improve this disorder by up-regulating probiotics and down-regulating the abundance of pathogenic bacteria.
- pulmonary fibrosis /
- Shuangshen Pingfei formula /
- gut microbiota /
- 16S rDNA /
- ROC analysis /
- Fusobacterium

HTML全文

图 1 双参平肺方对大鼠肺组织病理学的影响(×200)

Figure 1. Lung histological analysis of BLM-treated pulmonary fibrosis rats (×200)

下载: 全尺寸图片幻灯片

图 2 双参平肺方对PF大鼠肺组织HYP含量及MMP7 mRNA水平的影响

注: 与空白对照组比较, ^{* *}P < 0.01;与模型组比较, ^##P < 0.01。x±s, n=6。

Figure 2. HYP content and MMP7 mRNA levels in lung tissue of rats in each group

下载: 全尺寸图片幻灯片

图 3 多样性分析和物种组成分析

注: A.α多样性分析; B.β多样性分析; C.门水平和属水平物种组成分析; Control.空白组; Model.模型组; SSPF.双参平肺方组

Figure 3. Diversity analysis and species composition analysis

下载: 全尺寸图片幻灯片

图 4 物种差异与标志物分析

注: A.韦恩图; B.LEfSe分析; C.门水平和属水平物种组成热图; Control.空白组; Model.模型组; SSPF.双参平肺方组

Figure 4. Species difference and marker analysis

下载: 全尺寸图片幻灯片

图 5 不同菌属与生化指标的相关性

注: 红色代表正相关, 蓝色代表负相关, 颜色深浅表示相关性强弱; ^*P < 0.05, ^{* *}P < 0.01。

Figure 5. Correlation between different bacterial genera and biochemical parameters(Spearman analysis)

下载: 全尺寸图片幻灯片

图 6 代表性差异肠道菌群ROC曲线

注: A.空白组/模型组；B.模型组/双参平肺方组

Figure 6. ROC curves of representatively different gut microbiota

下载: 全尺寸图片幻灯片

表 1 双参平肺方对PF大鼠肺组织病理变化的影响(x±s, n=6)

Table 1. HE and Masson staining scores in lung tissue of rats in each group(x±s, n=6)

组别	Masson评分	HE评分
空白对照组	0.33±0.26	1.50±1.64
模型组	2.33±0.52^**	6.17±0.75^**
双参平肺方组	1.25±0.61^##	3.67±0.82^##
注: 与空白对照组比较, ^{* *}P < 0.01;与模型组比较, ^##P < 0.01。

下载: 导出CSV

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