续苓健骨方对骨质疏松模型大鼠肠道菌群结构和功能的影响

Effects of Xuling Jiangu Recipe on the Composition of Intestinal Flora and Gene Function in Osteoporosis Model Rats

摘要: 目的：研究续苓健骨方对去卵巢骨质疏松模型大鼠肠道菌群的组成及基因功能影响。方法：建立去卵巢骨质疏松大鼠模型，随机分为四组：假手术组、模型组、续苓健骨方组、雌激素组，每组6只。造模4周后，药物干预12周取材，双能X线骨密度仪小动物扫描模式检测大鼠胫骨骨密度，宏基因组全基因组测序技术检测大鼠肠道菌群的组成、结构及功能。结果：与假手术组相比，模型组大鼠胫骨骨密度显著降低(P < 0.01)；与模型组比较，续苓健骨方组和雌激素组骨密度明显提高(P < 0.01)；对不同组别肠道菌群的物种组成及丰度情况进行分析，在门、纲、目、科、属、种各个水平，组间差异有统计学意义(P < 0.001)；LEfSe差异性分析结果显示，厚壁菌门、杆菌纲、乳酸杆菌目等菌种是骨质疏松症模型组大鼠肠道内关键差异菌群；与模型组比较，古菌门、甲烷杆菌纲目科、甲烷球菌属种是续苓健骨方组关键差异菌群；进一步对差异基因的KEGG功能注释结果统计并进行富集分析，在KEGG第一层级水平，各组大鼠肠道菌群基因注释到6大类生物代谢通路，其中注释到第一位的为新陈代谢通路；在KEGG第二层级水平，注释到的主要代谢通路包括：碳水化合物代谢、氨基酸代谢等，富集到的通路中60%属于在KEGG一级水平中的新陈代谢通路；对基因注释得到的功能丰度进行差异分析，续苓健骨方组与骨质疏松症组肠道菌群基因功能注释结果差异具有统计学意义，其中差异明显的代谢通路包括内分泌抵抗通路、ErbB信号通路、TGF-β信号通路等。结论：续苓健骨方在骨质疏松模型大鼠肠道菌群结构组成、多样性及生物代谢通路上具有调节作用，平衡肠道微生态稳态是续苓健骨方实现防治骨质疏松症作用的靶点之一。

Abstract: Objective: To study the effect of Xuling Jiangu recipe on the composition and gene function of in-testinal flora in ovariectomized osteoporosis model rats. Methods: Rat models of ovariectomized osteoporosis were established and randomly divided into four groups: sham operation group, model group, Xuling Jiangu recipe group, and estrogen group, with 6 rats in each group. After 4 weeks of modeling, samples were collected after 12 weeks of drug intervention. The bone mineral density of rat tibia was detected by dual-energy X-ray absorptiometry in small animal scanning mode, and the composition, structure and function of intestinal flora in rats were detected by metagenomic whole genome sequencing technology. Results: Compared with the sham operation group, the bone mineral density of the tibia of the model group was significantly decreased (P < 0.01); The species composition and abundance of intestinal flora in different groups were analyzed, and there were statistically significant differences between groups at the levels of phylum, class, order, family, genus, and species (P < 0.001); LEfSe difference analysis results. It showed that the bacteria species such as Firmicutes, Bacillus, Lactobacillus were the key differential flora in the intestinal tract of rats in the osteoporosis model group; It is the key differential flora of the Xuling Jiangu recipe group; further statistics on the KEGG functional annotation results of differential genes and enrichment analysis are carried out. At the first level of KEGG, the intestinal flora genes of rats in each group are annotated into 6 categories of organisms Metabolic pathways, of which the first annotated is the metabolic pathway; at the second level of KEGG, the main metabolic pathways annotated in-clude: carbohydrate metabolism, amino acid metabolism, etc. 60% of the enriched pathways be-long to the KEGG level Metabolic pathways in the horizontal level; the difference analysis of the functional abundance obtained by gene annotation, the results of functional annotation of intes-tinal flora genes between the Xuling Jiangu prescription group and the osteoporosis group were statistically significant, and the metabolic pathways with significant differences were significantly different including endocrine resistance pathway, ErbB signaling pathway, TGF-β signaling pathway, etc. Conclusion: Xuling Jiangu recipe has a regulatory effect on the composition, diversity and biological metabolic pathways of intestinal flora in osteoporosis model rats. Balancing the intestinal microecological homeostasis is the key to the prevention and treatment of osteoporosis by Xuling Jiangu recipe, which is one of the targets of the disease.