Microbiology Spectrum:“强大助攻”益生菌!可增强哮喘常规用药治疗效果!

2021-10-18 MedSci原创 MedSci原创

Microbiology Spectrum:补充益生菌能通过调节肠道微生物组和血清代谢组减轻哮喘症状

哮喘是一种常见的慢性炎症性呼吸系统疾病,在全球范围内具有高发病率和死亡率,其临床表现为喘息、咳嗽、胸闷、呼吸急促、呼吸困难、睡眠障碍等,并且可能诱发气胸、肺不张、慢阻肺、支气管扩张或肺源性心脏病等并发症。
 
目前有3.3亿儿童和成人正在遭受哮喘的折磨,预计到2025年患有哮喘的人数将增加到4亿。哮喘的发病机制并不明确,目前研究表明肠道失调很可能是诱发这类过敏性疾病的重要因素之一。本研究为一项为期3个月的随机对照试验,研究乳酸双歧杆菌M8 (Probio-M8)在两个支气管哮喘队列(益生菌组,n = 29;安慰剂组26例)的作用。Probio-M8是一种从健康女性母乳中分离的益生菌菌株,该菌株在之前的研究中已显示出多种促进健康的特性。

本研究旨在通过分析患者的临床指标、肠道菌群和血清代谢组的变化,评价m8益生菌缓解哮喘的临床疗效,并揭示其缓解哮喘症状的机制,为哮喘的发病机制和治疗、新型疾病管理策略的机制以及益生菌治疗的应用提供新的见解。

1、益生菌可改善哮喘相关症状。

为了评估益生菌治疗哮喘症状的辅助疗效,在第0、30和90天监测了多个临床参数,包括哮喘对照试验评分(ACT)、CaNO、呼气峰流量(PEF)、PEV1和FeNO,在第0天,益生菌组和安慰剂组在这些指标上没有观察到显著差异。

有趣的是,与安慰剂组相比,Probio-M8联合布地奈德/福莫特罗吸入剂在干预第30天显著降低FeNO水平(P = 0.049),并在干预结束时改善ACT评分(P = 0.023)。更重要的是,Probio-M8组在干预第30天显著降低了CaNO水平(P = 0.038),干预90天后这种效果更加明显(P = 0.001)。

此外,与第0天相比,干预前后Probio-M8组ACT评分显著升高,CaNO和FeNO水平降低(P < 0.05)。Probio-M8组与安慰剂组在肺功能指标(PEF、PEV1、PVC)和外周血嗜酸性粒细胞计数方面均未观察到辅助疗效(表S3)。m8益生菌组与安慰剂组各时间点血清IgE水平无显著差异;而安慰剂组的IgE水平在研究期间和研究结束时显著升高(P <0.003;图1 a)。提示m8益生菌辅助治疗可明显改善部分哮喘相关临床症状。

2、基因组的特性

在第0、30和90天,对31名受试者共93个样本进行了深入的MAGs分析。结果表明大多数基因组属于已知的微生物群落(图1b)。跟踪样品中的Probio-M8发现在益生菌组中,Probio-M8菌株的丰度逐渐增加,而在安慰剂组中则没有,这表明摄入的Probio-M8菌株可以很容易地通过消化道(图S2c)。

3、益生菌调节肠道菌群组成

Probio-M8组和安慰剂组在各时间点的α多样性和β多样性均无显著差异(P> 0.05,图2a和b)。然而,与第0天相比,安慰剂组的α多样性在第30和90天显著降低(P值分别为0.05和0.005),而Probio-M8组在整个试验期间保持稳定(图2a)。这表明摄入益生菌可能有助于维持肠道菌群多样性的稳定。此外,按年龄进行亚组比较时,各时间点的肠道菌群α / β多样性均无显著差异(成人:28 ~ 59岁;老年人:61岁至71岁;P >0.05;图S2d和e)。

 

进一步探索肠道菌群变化,发现在第0天两组间丰度没有显著差异,但在时间点之间或处理组之间丰度有差异(表S6)。对于益生菌组,三种SGB——T16C.M023 (Bifidobacterium animalis), C14B.M022 (Roseburia hominis), 和C1A.M016 (Ruminococcus callidus)显著增加,而1种SGB ——C5A.M066,(Parabacteroides distasonis)显著降低(P < 0.05)。安慰剂组检测到16种应答性SGBs,包括C8B.M011,T17A.M023,C12C.M016, T5B.M042, T16A.M006(梭状芽胞杆菌科),T10B.M022, C6C.M025, T4C.M019(厚壁菌门)和T4C.M040, C14C.M035,干预期间/干预后的丰度变化显著。
三个SGBs在/干预后明显减少,而三个SGBs则显著升高(P< 0.05)。在干预结束时,三种益生菌响应性SGBs在组间表现出差异丰度。在第90天,比起安慰剂组,Probio-M8组中C12A.M023(长双歧杆菌)和T2B.M023(普雷沃氏菌sp. CAG)显著增加,而C8C.M022(梭菌属的细菌)则相反。(P<0.05;图2 c)。
3、益生菌调节GMMs和预测肠道生物活性化合物
利用MetaCyc和KEGG数据库进行了以基因组为中心的代谢重建,主要关注与哮喘相关的发展、病理生理学和免疫应答相关的模块(表S7)。共鉴定出10个不同门,主要为厚壁菌门(72.75%)、拟杆菌门(13.11%)和放线菌门(4.11%)。有趣的是,在干预期间/之后,安慰剂组和益生菌组之间的分布有很大的不同。两种GMMs,丁酸合成I和色氨酸降解,在益生菌组中占主导地位,而醋酸盐降解在安慰剂组中占很高比例(图3a)。
MelonnPan进一步预测了肠道活性化合物,共鉴定出80个代谢产物。Procrustes分析结果证实了肠道微生物组与代谢组之间具有良好的协同性(相关系数= 0.351;P = 0.001),表明在治疗过程中,预测的肠道代谢物和SGB之间的黏结性改变(图3b)。13种反应性代谢物在第0天没有显著变化,但在治疗期间/之后在安慰剂组和益生菌组之间出现了显著差异(表S8)。在第30天和第90天,益生菌组的肾上腺酸、C18:0鞘磷脂、C18:1胆固醇酯、红嘌呤酸和亚麻油酰乙醇酰胺的含量显著增加(P<0.05;图3c),代表了m8在促进特定生物活性代谢物及相关通路合成方面的辅助效果。
4、益生菌调节血清代谢物
第30天时,益生菌组与安慰剂组血清代谢组差异无统计学意义(P < 0.06)然而,通过PCoA和Adonis检验对血清代谢组进行纵向分析时,发现Probio-M8联合布地奈德/福莫特罗吸入剂组在第30天和第90天的血清代谢组发生了接近显著的变化(P值分别为0.06和0.04;Adonis试验),但安慰剂组未见类似变化(图4a)。在第90天,与安慰剂组相比,益生菌组的6个特征,即5-十二烯酸、肠二醇、丁香酸、1-棕榈酰乙酸-甘油、色氨酸和鞘磷脂都明显增强(图4c)。提示Probio-M8联合布地奈德/福莫特罗吸入剂在治疗过程中特异性调节了部分血清代谢物。
5、益生菌调节肠道病毒谱
研究结果表明:肠道病毒群具有较高的新颖性(图5a)。在所有时间点上,Probio-M8组和安慰剂组肠道病毒群的α和β多样性均无显著差异(P>0.05;图5b和c)。然而,安慰剂组肠道病毒群的多样性显著降低(P< 0.05),而益生菌组无此变化(图5b),说明Probio-M8+布地奈德/福莫特罗吸入剂联合用药有助于维持肠道噬菌体的多样性。PCoA发现,在不同时间点,安慰剂组和益生菌组的整体肠道噬菌体谱没有显著差异(P >0.05, ANOSIM;图5 c)。
在我们的数据集中鉴定了10个噬菌体科,其中最占优势的病毒科是Iridoviridae和Podoviridae(图5d)。Probio-M8组Myoviridae的平均丰度在第30天显著降低,而Herelleviridae的平均丰度在第90天显著升高(图S3a)。值得注意的是,肠道细菌菌群的α多样性与肠道病毒群的α多样性具有很强的相关性(总体R = 0.895, P< 0.001,图5 e;益生菌组R = 0.916, P < 0.001 ;安慰剂组 R = 0.873, P < 0.001)。Procrustes分析一致发现肠道细菌微生物群和肠道病毒群之间存在强相关(相关系数= 0.782;P = 0.001;图5f),提示肠道病毒对其细菌宿主具有高度的感染特异性。与安慰剂组相比,益生菌组的肠道菌群和肠道病毒群之间的互动连接网络更为强大(图S3b和c)。

6、肠道菌群对监测特征的影响大小
结果显示,在益生菌组和安慰剂组之间,272个SGBs (P < 0.05)对纳入特征的方差平均贡献了55.34%(范围从28.8%到78.31%)。值得注意的是,与安慰剂组相比,益生菌组的肠道菌群对预测生物活性化合物、临床指标和血清代谢物的解释差异分别为9.80%、3.60%和1.30%(图6a)。
另一方面,在噬菌体宏基因组中观察到相反的趋势。有20种sbs对probioo - m8处理始终表现出最强的效应量,特别是长双歧杆菌(C12A.M023)、厚壁菌门细菌AM10-47 (T10B.M022)、瘤胃球菌sp. AF37-6AT (T17B.M018)和厚壁菌门细菌CAG:83 (T4C.M019)的sbs。在干预结束时,益生菌组富集了这些蛋白质(图S3d)。综上所述,与单纯常规药物治疗相比,益生菌+布地奈德/福莫特罗吸入剂联合用药对临床症状、血清代谢组和肠道微生物代谢潜力的影响更大,对肠道病毒群的影响较小。
综上,评估益生菌在控制哮喘症状方面的辅助功效试验结果发现,益生菌作为哮喘治疗辅助剂可以显著提升治疗效果。通过研究哮喘患者的肠道微生物多样性,发现摄入益生菌增加了肠道微生物群的恢复能力,使肠道微生物多样性水平维持在基线水平,说明益生菌的联合使用通过维持肠道微生物群多样性的稳定,还能够使得有益菌群富集,缓解哮喘。并且益生菌能够增强抗炎微生物的生物活性潜力,进而促进哮喘治疗功效。
除此之外,益生菌组受试者血液中十二烷酸、肠二醇、色氨酸以及鞘磷脂等代谢物水平提升,而这些代谢物能够发挥抗炎作用参与免疫反应,说明益生菌能够作为宿主和肠道微生物之间的调节因子,在联合常规治疗哮喘的过程中能特异性地调节部分血清代谢物,提高治疗效果。
总的来说,益生菌对缓解哮喘症状有一定的作用。益生菌与常规药物治疗联合应用可维持哮喘患者肠道细菌菌群和病毒群的多样性和稳定性。同时,益生菌的应用通过增加某些抗炎血清代谢物、肠道生物活性代谢物及相关途径,对哮喘患者产生了促进健康的作用。这些变化直接调节肠-肺轴,提高了治疗效果。本研究结果表明,联合使用Probio-M8与常规治疗协同缓解肠道-肺轴相关疾病,如哮喘,扩大了相关慢性疾病的治疗选择。
原文来源:
Ailing Liu, et al.Adjunctive Probiotics Alleviates Asthmatic Symptoms via Modulating the Gut Microbiome and Serum Metabolome.Microbiology Spectrum,September/October 2021  Volume 9  Issue 2  e00859-21

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    2022-04-25 sunylz
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    2022-02-10 xjy02
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    2022-06-19 feather89

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