Analysis Of Intestinal Flora Diversity In Patients With Primary Osteoporosis Of "spleen And Kidney Yang Deficiency" Typ

Aim:To explore the structural characteristics of the gut microbiota in patients with primary osteoporosis of "spleen-kidney yang deficiency" type,and to study the mechanism of gut microbiota on "spleen-kidney yang deficiency" primary osteoporosis by comparing the differences of gut microbiota structure and metabolites between patients with "spleen-kidney yang deficiency" primary osteoporosis and normal healthy people.Material and method:In this study,a case-control study method was used to select socially healthy subjects and the subjects who were treated in the First Orthopaedic Ward of the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine from April2021 to November 2021.According to the diagnostic criteria of TCM syndrome and osteoporosis,they were divided into normal group and spleen-kidney yang deficiency osteoporosis group.After fully explaining the purpose and test method of this study with the subjects and their families,and signing the informed consent,the basic information such as age,gender and eating habits of the two groups of subjects were counted by questionnaire.The venous blood of the two groups of subjects was collected,and the related indexes of bone metabolism were detected and counted.The feces of two groups of subjects were collected,the genomic DNA of gut microbiota was extracted,the DNA of 16sv3-v4 region of gut microbiota was amplified by PCR,and the original sequence was obtained.The original sequence is de cluttered to generate the optimized sequence,and the optimized sequence is classified into multiple operation units according to the similarity.After comparing each operation unit with the gene bank,the species annotation is obtained.Combined with the species annotation,the structure of the two groups of gut microbiota is analyzed.This study uses spss22 0 software for statistical analysis of data.Results:In this study,43 subjects were collected,2 cases fell off,and 41 cases were finally included.There were 22 subjects in the normal group and 19 subjects in the spleen-kidney yang deficiency osteoporosis group.There was no significant difference in gender,β-CTX、PINP、IL-10 and OCN between the two groups;PTH and VD3 in the spleen-kidney yang deficiency osteoporosis group were lower than those in normal group(P < 0.05).The gut microbiota OTUs of the two groups partially overlapped.According to the analysis of alpha diversity between the two groups,there is a significant difference in alpha diversity index between the two groups.Chao1 index and observed species index in osteoporosis group are higher than those in normal group,indicating that there are more species of gut microbiota in osteoporosis group than in normal group.According to the principal component analysis of the two groups of gut microbiota,there are different aggregation tendencies,and the structural composition of gut microbiota is different;The structure analysis of gut microbiota at each classification level of the two groups is displayed in the form of relative abundance histogram.The analysis of different species between the two groups shows that the structure of gut microbiota was different between the two groups at each classification level.In the comparison of main gut microbiota at each classification level,it is found,at the target level,that the abundance of Burkholderiales in the osteoporosis group is lower than that in the normal group.At the genus level,the abundance of Blautia in osteoporosis group was lower than that in normal group,at the species level,that the abundance of Bacteroides vulgatus,Bacteroides stercoris,Bacteroides coprocola in the osteoporosis group is lower than that in the normal group,with statistical difference.The difference of main gut microbiota between the two groups at other classification levels was not significant after statistical analysis;In the low abundance gut microbiota at each taxonomic level,there are many different gut microbiota between the two groups,including spirochaetota et al.Lefse analysis showed that the key biomarkers with statistical difference between the two groups were lactobacillales,agathobacter,blautia,lachnoclostridium,lachnospiraceae,lachnospirales,parasutterella,sutterellaceae and burkholderiales.Conclusion:1.In this experiment,the number of gut microbiota species in patients with primary osteoporosis of spleen-kidney yang deficiency type was significantly different from that in normal healthy people(P < 0.05).2.There were differences in the structure of gut microbiota between the two groups.Among the main gut microbiota with high relative abundance(≥ 1%),the relative abundance of Burkholderiales in osteoporosis group was lower than that in normal group at the order level,the relative abundance of Blautia in osteoporosis group was lower than that in normal group at the genus level,and the relative abundance of Bacteroides vulgatus,Bacteroides stercoris,Bacteroides coprocola in the osteoporosis group was lower than that in the normal group at the species level.At the phylum,class and family,no significant differences were found in gut microbiota with higher relative abundances.In the gut microbiota with low relative abundance,there are many species with significant differences at each classification level.3.The species that play a key role in the structural difference of gut microbiota between the two groups of subjects include agathobacter,blautia,lachnoclostridium,lachnospiraceae,lachnospirales and parasutterella.Most of these key differential species can produce SCFAs,so the gut microbiota may affect the occurrence and development of primary osteoporosis of spleen-kidney yang deficiency type through SCFAs.