补肾法对绝经后骨质疏松症肠道菌群的影响

Effect of kidney-tonifying therapy on intestinal flora of postmenopausal osteoporosis

目的探讨补肾法对绝经后骨质疏松症的可能作用靶点。方法将50只雌性大鼠随机分为空白组、模型组、青娥丸组、补肾化痰组和利维爱组,各10只。采用双侧卵巢切除术制备绝经后骨质疏松症模型。术后第5周开始灌胃,持续12周后取各组大鼠股骨标本,采用micro-CT检测大鼠骨密度及骨微结构。每组取5只大鼠结肠粪便样本,在样本质量评价后每组纳入3个样本进行数据质控,对物种组成、相对丰度信息、主成分和Network网络相关性等进行分析。结果补肾法代表方青娥丸和补肾化痰组均可增加去势骨质疏松大鼠模型骨密度,抑制骨丢失。15个样本中共含有细菌2023种、206种古菌、227种真菌、2508种病毒、以及1035种质粒基因组序列,并按照相对丰度比例显示了排在前20的物种。热图聚类显示各组组内菌属的分布模式规律存在一定相似性;Anosim分析R值为0.201,P值为0.0855;主成分分析提示所有样本的物种相似度较高,组间物种存在不同。说明了PMOP模型大鼠肠道菌群显示了一定的物种多样性,在已有物种属水平基础上,进一步进行Network网络分析发现相关性>0.5(P<0.05)的物种有37种,与丰度排于前20的物种比对后,发现补肾法药物组干预后PMOP可靠菌属为:乳酸杆菌属(Lactobacilles),约氏乳酸杆菌、罗伊氏乳酸杆菌、瑞士乳酸杆菌和卷曲乳酸杆菌尤甚。结论肠道菌群可能是补肾法治疗PMOP的关键靶点之一,其中乳酸杆菌属可能是重要的研究切入点。

Objective To explore the possible target of tonifying,the kidney on postmenopausal osteoporosis.Methods 50 female rats were randomly divided into blank group,model group,Qing e Pill Group,Bushen Huatan Group and Lévis group,10 each.A model of postmenopausal osteoporosis was established by bilateral Oophorectomy.At the 5 th week after operation,the rats femur specimens were taken after 12 weeks,and the bone mineral density and bone microstructure were measured by micro-CT.Five fecal samples were collected from each group,and three samples were included in each group for data quality control.Species composition,relative abundance,principal component and Network correlation were analyzed.Results BMD and bone loss in ovariectomized rats were increased by kidney-tonifying method on behalf of Fangqing’e Pill and kidney-tonifying and phlegm-resolving group.The 15 samples contained 2,023 bacterial species,206 ARCHAEA,227 fungi,2,508 viruses,and 1,035 plasmid genome sequences,showing the top 20 species in relative abundance ratio.The results of thermographic cluster analysis showed that the distribution pattern of Genera in each group was similar,the R value of Anosim analysis was 0.201,the P value was 0.0855,and the principal component analysis indicated that the species similarity of all samples was high,and there were different species among groups.The results showed that the intestinal microflora of PMOP model rats showed a certain species diversity,and there were 37 species with correlation>0.5(P<0.05),compared with the top 20 species in abundance,the reliable PMOP strains were lactobacillus,lactobacillus,lactobacillus,lactobacillus Reuteri,lactobacillus Helveticus,lactobacillus helveticus and Lactobacillus Crimulus in the group of tonifying kidney.Conclusion Intestinal Flora may be one of the key targets of kidney-tonifying therapy for PMOP,and lactobacillus may be an important research entry point.