Investigation Of Therapeutic Mechanism Of Compatibility Of Aconiti Lateralis Radix Praeparata And Paeoniae Radix Alba In Wind-Cold-Dampness Type Bi-Syndrome

Objective:Based on the theory of compatibility of traditional Chinese medicine,combined with the wind-cold-dampness type bi-syndrome model rats,the analgesia,anti-inflammatory and acute toxicity experiments,calculation of combination index,determination of inflammatory cytokines,histopathological examination of synovium and metabolomics were used as the main technical means to evaluate the efficacy enhancing and toxicity reducing effects and its scientific connotation of compatibility of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in the treatment of wind-cold-dampness type bi-syndrome.Methods:1.Using the body-twisting test induced by acetic acid and the pain test induced by hot-plate,rapidly select the main components of alkaloids of Aconiti Lateralis Radix Praeparata that could produce analgesic effect.2.The analgesic and anti-inflammatory effects of compatibility of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in the treatment of wind-cold-dampness type bi-syndrome were studied by the joint pressing pain and foot swelling experiments.The best compatibility components and proportion of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba was selected for the study of the efficacy enhancing mechanism by combination index,cytokines,histopathology and metabolomics.3.Acute toxicity experiment was used to study the toxicity reducing effect of compatibility of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba,and the best compatibility proportion was selected for the study of the toxicity reducing mechanism by metabolomics.4.Based on serum anti-cyclic citrullinated peptide antibody,the combination index of compatibility of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in the treatment of wind-cold-dampness type bi-syndrome was calculated by Ting-Chao Chou method and CompuSyn software.The efficacy enhancing effects of the two drugs were investigated by the serum contents of IL-1?,IL-6,IL-10,TNF-?,IFN-?and HE staining results of synovial tissue.5.UPLC-LTQ/Orbitrap MS with untargeted metabolomics,combined with multivariate statistical analysis including PCA and PLS-DA,the differential endogenous metabolites were screened and identified.The efficacy enhancing and toxicity reducing mechanism of compatibility of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in the treatment of wind-cold-dampness type bi-syndrome was analyzed from the perspective of small molecule metabolite changes.Results:1.Lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata could significantly improve the pain tolerance of mice induced by acetic acid and hot-plate,which were the main components that produce analgesic effect.2.The compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony or polysaccharide of peony had better analgesic and anti-inflammatory effects in the treatment of wind-cold-dampness type bi-syndrome,and the compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony?proportion of medicinal materials 1:2?was the best choice.3.The LD₅₀ values of compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony were significantly higher than that of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata,which had better toxicity reducing effect,and the best compatibility proportion was 1:2.4.The combination index of compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony?proportion of medicinal materials 1:2?was 0.213,indicating synergistic effect in the treatment of wind-cold-dampness type bi-syndrome.The serum contents of IL-1??IL-6?TNF-??IFN-?in wind-cold-dampness type bi-syndrome model rats were significantly higher than healthy rats,with severe synovial hyperplasia accompanied by a large number of inflammatory cells infiltration.Compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony?proportion of medicinal materials 1:2?had significant inhibitory effects on serum IL-1?,IL-6 and IFN-?,and had obvious therapeutic effects on synovial hyperplasia and inflammation,the therapeutic effects were better than that of single drug administration groups.5.Compared with lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata administration group,compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony could enhance the regulation of citric acid,tryptophan,phenylalanine and catechol metabolic disorders in wind-cold-dampness type bi-syndrome rats,and had unique regulatory effects on the metabolic disorders of tyrosine,vanillylmandelic acid and isoleucine,but had no obvious regulatory effects on the metabolic disorders of leucine,arginine and linoleate.Total glucosides of peony could regulate the metabolic disorders of galactitol,6-hydroxynicotinic acid,adenosine,carnitine,arginine,UMP and lysine induced by the toxic dose of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata,to produce detoxification effects,but it had no obvious regulatory effects on the metabolic disorders of D-galactose,deoxycholic acid,glycochenodeoxycholic acid,proline,N-formylanthranilic acid and 2-Deoxycytidine.Conclusions:Compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony in the treatment of wind-cold-dampness type bi-syndrome had better analgesic and anti-inflammatory effects,and could significantly reduce the overexpression of some pro-inflammatory cytokines such as IL-1??IL-6?IFN-?in vivo,prevention of synovial lesions,enhance the regulatory effects on energy and amino acid metabolic disorders to achieve synergistic effects.The compatibility of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata and total glucosides of peony could significantly improve the tolerance of wind-cold-dampness type bi-syndrom rats to the toxicity of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata,and total glucosides of peony could reduce the interference of lipid-soluble alkaloids of Aconiti Lateralis Radix Praeparata on fat,amino acids and energy metabolism,which was a possible mechanism of toxicity reducing.