双参平肺方对博来霉素致肺纤维化大鼠肠道菌群的影响
Effect of Shuangshen Pingfei Formula on Gut Microbiota in BLM-Induced Pulmonary Fibrosis Rats
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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。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.
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