Effect Of Reinforcing Qi And Nourishing Blood Formula Prescription On Intestinal Flora Of Knee Osteoarthritis(Qi And Blood Deficiency Type)

Objective:To investigate the efficacy of Reinforcing qi and nourishing blood formula prescription on patients with osteoarthritis of the knee(KOA)(Qi and Blood deficiency type)and the effect of intestinal flora,so as to provide a theoretical basis for the follow-up study on the prevention and treatment of osteoarthritis of the knee by Chinese medicine intervention on intestinal flora.Methods: A total of 20 patients with osteoarthritis of the knee(Qi and Blood deficiency type)were included as the treatment group and 20 healthy volunteers were included as the control group.The treatment group was treated with reinforcing qi and nourishing blood formula prescription for 8 weeks,and the imaging classification,joint function classification,VAS score,WOMAC score,Lequesne score and TCM syndrome score of KOA patients were compared before and after treatment.Stool specimens from KOA patients before treatment were collected as the pre-treatment group,stool specimens after treatment were collected as the post-treatment group,and stool specimens from 20 healthy volunteers were collected as the healthy control group,a total of 60 samples were collected and sequenced based on the Pac Bio Sequel II platform.The sequencing results of the three groups were subjected to categorical operational unit statistics,Alpha and Beta diversity analysis,between-group difference significance analysis to compare the differences in intestinal flora between the healthy and pre-treatment groups and between the pre-treatment and post-treatment groups,and Lef Se analysis to identify biomarkers that were significantly different after the three groups.Results: 1.The imaging classification of KOA patients decreased after treatment with reinforcing qi and nourishing blood formula prescription,but it was not statistically significant(P>0.05);the joint function classification,VAS score,WOMAC score,Lequesne score and TCM evidence score all decreased and were statistically significant(P<0.05).2.A total of 749 OTUs were obtained in the three groups.The number of OTUs increased after treatment with reinforcing qi and nourishing blood formula prescription,and the number of OTUs overlapping with the healthy control group increased in the post-treatment group.3.Alpha diversity analysis showed that the sequencing coverage of sample species was reasonable and the sequencing amount was sufficient.The richness and diversity of the pre-treatment group were lower than those of healthy people,with significant difference in richness(P < 0.05)and no significant difference in diversity(P >0.05).The diversity of the group after treatment was lower than that before treatment,but there was no statistical significance(P > 0.05).4.Beta diversity analysis: PC1 explained24.61% and PC2 explained 14.60% in PCo A analysis,both healthy control and pre-and post-treatment groups had their own tendency to aggregate,and the post-treatment group had the highest degree of aggregation.n MDS analysis Stress < 0.2.Statistical tests:PERMANOVA test: R2 = 0.103,P = 0.001;Anosim test: R = 0.160,P = 0.001.5.The groups with significant differences between groups were analysed at each taxonomic level: the most significant difference at the phylum level was Firmicutes;the greatest difference in abundance at the order level was Bacteroidia;the species with significant differences at the order level was Bacteroidales;the greatest difference in abundance at the family level was Bacteroidaceae;at the genus level Bacteroides The greatest differences were found at the family level for Bacteroidaceae;at the genus level for Bacteroides the most diverse;and at the species level for Megamonas funiformis.6.LEf Se analysis showed that the most representative biomarkers in the healthy control group were Firmicutes,Bacilli,Peptostreptococcaceae,Bacteroidaceae,Bacteroides,Dialister invisus;the most representative biomarkers in the post-treatment group were Selenomonadaceae,Megamonas,Megamonas funiformis.Conclusions: 1.The flora abundance of KOA(Qi and blood deficiency type)patients and healthy volunteers was different in phylum,class,order,family,genus and species,and some of the flora abundance was corrected after treatment,which explains that reinforcing qi and nourishing blood formula prescription may intervene in the occurrence and development of KOA(Qi and blood deficiency type)by regulating intestinal flora.2.After treatment,some microflora did not improve,which provided a reference for clinical adjustment of the use of prescription.