基于高通量测序技术研究归脾汤对心脾两虚型轻度阿尔茨海默病患者肠道菌群多样性的影响

Effect of Guipi Tang on Intestinal Flora Diversity in Patients with Mild Alzheimer Disease with Deficiency of the Heart and Spleen Type Based on High-Throughput Sequencing Technology

目的:研究归脾汤对心脾两虚型轻度阿尔茨海默病(AD)患者肠道菌群的影响,探讨归脾汤改善心脾两虚型轻度AD与肠道菌群的关系。方法:选取12例心脾两虚型轻度AD患者的粪便样品作为研究对象,包括服用归脾汤前的样本及连续服用归脾汤1个月后的样本,观察提取各研究样本的染色体DNA,并进行DNA片段长度检测和定量。对提取的DNA样本进行16S r RNA V3区基因扩增及焦磷酸测序。结果:归脾汤给药1个月后,AD患者的肠道菌群整体结构与给药前有统计学差异,主要表现为厚壁菌门、拟杆菌门、变形菌门、放线菌门和互养菌门丰度降低。在Cell division、Ether lipid metabolism这2个代谢通路中,给药前组与给药后组的平均丰度比较,差异均有统计学意义(P<0.05)。给药后组微生物在Cell division通路中的平均丰度高于给药前组(P<0.05),在Ether lipid metabolism通路中的平均丰度低于给药前组(P<0.05)。结论:肠道菌群结构的异常改变与AD的发生发展存在较强的关联性。归脾汤加减可能是通过改变肠道菌群结构的机制改善心脾两虚证轻度AD患者的症状。

Objective:To study the effect of Guipi tang on intestinal flora in patients with mild Alzheimer disease(AD)with deficiency of the heart and spleen type,and to explore the relationship between Guipi tang and intestinal flora in improving mild AD with deficiency of the heart and spleen type.Methods:Stool samples from 12 patients with mild AD with deficiency of the heart and spleen type were selected as study subjects,including samples before taking Guipi tang and samples after continuously taking Guipi tang for a month.The DNA of chromosome in each sample was observed and extracted,and the length of DNA fragments was detected and quantified.The 16 S rRNA V3 region gene amplification and pyrosequencing were performed on the extracted DNA samples.Results:One month after the administration of Guipi tang,the overall structure of intestinal flora in AD patients was statistically different from that before administration,mainly manifested as decreased abundance of firmicutes,bacteroidetes,proteobacteria,actinomycetes and mycobacteria.In the two metabolic pathways of cell division and ether lipid metabolism,the average abundance of the patients before and after administration was statistically different(P<0.05).The average abundance of microbe in cell division pathway after administration was higher than that before administration(P<0.05),and the average abundance in the ether lipid metabolism pathway after administration was lower than that before administration(P<0.05).Conclusion:There is a strong correlation between the abnormal structure of intestinal flora and the occurrence and development of AD.Modified Guipi tang may be used to improve the symptoms of mild AD by changing the structure of intestinal flora.