绝经综合征患者肠道菌群结构变化及桂仙合剂的干预作用

Changes of Intestinal Flora Structure in Patients with Menopausal Syndrome and the Intervention Effect of Guixian Mixture

目的:该研究旨在探究绝经综合征(MPS)患者肠道菌群结构变化及中药桂仙合剂(GXD)干预后对肠道菌群结构的影响。方法:选取符合标准的MPS患者35名作为MPS组(E组,n=35),同时招募12名绝经前后健康女性作为健康组(H组,n=12),E组患者在GXD干预后作为治疗组(AE组,n=35)。使用16S rRNA测序对3组所采集的粪便标本进行肠道菌群测序分析及功能预测。结果:多样性分析显示,E组与AE组间存在显著差异(P<0.05),而E组与H组、AE组与H组间无差异(P>0.05)。与H组相比,E组厚壁菌门(Firmicutes)/拟杆菌门(Bacteroidetes)比值降低,但无差异(P>0.05);AE组该比值上升,并与E组存在显著差异(P<0.05)。在属水平上,经粘液真杆菌属(Blautia)、另枝菌属(Alistipes)在AE组中富集(P<0.05)基于线性判别分析(LEfSe)发现,直肠真杆菌属(Agathobacter)及经粘液真杆菌属(Blautia)对3组肠道菌群间显著性差异影响力最大(LDA score>4)。组间DESeq2显示,E组有6个菌属丰度显著高于H组,其中黄杆菌属(Flavonifractor)、霍尔德曼氏菌属(Holdemanella)在GXD干预后丰度上调;直肠真杆菌属(Agathobacter)、梭菌属(Clostridia_UCG_014)丰度显著低于H组,但在GXD干预后丰度上调。功能预测显示,E组在脂多糖生物合成功能上显著增强(P=0.03),但在GXD干预后减弱;而甲状腺激素信号传导通路、多巴胺能突触功能、逆行性的内啡肽信号传递功能、羟色胺能突触功能在GXD干预后显著增强(P<0.01)。结论:与绝经前后健康女性相比,MPS患者肠道菌群结构出现明显改变,而中药GXD可以调整其肠道菌群结构,并可能通过改善后的肠道菌群调整相关功能的强弱,进而改善MPS相关症状。

Objective:This study aims to investigate the changes in the structure of intestinal flora in patients with menopausal syndrome(MPS)and the effect of traditional Chinese medicine Guixian mixture(GXD)on the structure of intestinal flora after intervention.Methods:35 MPS patients who met the criteria were selected as the MPS group(group E,n=35),and 12 healthy women before and after menopause were recruited as the healthy group(group H,n=12).Treatment group(AE group,n=35).16S rRNA sequencing was used to perform sequencing analysis and functional prediction of intestinal flora on the stool samples collected from the three groups.Results:Diversity analysis showed that there was a significant difference between E group and AE group(P<0.05),but there was no difference between E group and H group,AE group and H group(P>0.05).Compared with group H,the ratio of Firmicutes/Bacteroidetes in group E decreased,but there was no difference(P>0.05);the ratio increased in group AE,and there was a significant difference from group E(P<0.05).At the genus level,the genus Blautia and Alistipes were enriched in the AE group(P<0.05).Based on linear discriminant analysis(LEfSe),it was found that the genus Agatho bacterium and Blautia had the greatest influence on the significant differences among the three groups of intestinal flora(LDA score>4).DESeq2 between groups showed that the abundance of 6 genera in group E was significantly higher than that in group H,among which the abundance of Flavonifractor and Holdemanella were up-regulated after GXD intervention;(Agathobacter),Clostridia(Clostridia_UCG_014)abundance was significantly lower than that of group H,but the abundance was upregulated after GXD intervention.Functional prediction showed that group E significantly enhanced lipopolysaccharide biosynthesis function(P=0.03),but attenuated after GXD intervention;while thyroid hormone signaling pathway,dopaminergic synapse function,retrograde endorphin signaling function,serotonergic synapse function was significantly enhanced after GXD intervention(P<0.01).Conclusion:Compared with healthy women before and after menopause,the intestinal flora structure of MPS patients is significantly changed,and the traditional Chinese medicine GXD can adjust the intestinal flora structure,and may adjust the strength of related functions through the improved intestinal flora,Thus improving MPS-related symptoms.