Study On The Model Of Tripterygium Wilfordii-astragalus In The Treatment Of Collagen-induced Arthritis

Objective:Based on the literature database,we excavated the Tripterygium wilfordii-astragalus drug pair for the treatment of rheumatoid arthritis,and explored the possible mechanism of the effective components of the drug pair(calycosin,triptolide)in the treatment of collagen-induced arthritis in mice.To provide a preliminary basis for the development of new drugs by two joint applications in the later stage.Method:1.Based on the literature database,the combination of Tripterygium wilfordii and Astragalus membranaceus in the treatment of rheumatoid arthritis was analyzed.(1)all clinical prescriptions in Chinese Biomedical Literature Database from 1998 to2018 were searched by using "Tripterygium wilfordii","rheumatoid" and "test" as key words.(2)the compatibility law was analyzed by the auxiliary system of traditional Chinese medicine inheritance,and the combination prescription of Tripterygium wilfordii and Astragalus membranaceus was excavated.(3)the compatibility law was analyzed by the auxiliary system of traditional Chinese medicine inheritance,and the combination prescription of Tripterygium wilfordii and Astragalus membranaceus was excavated.2.Treatment of collagen-induced arthritis by regulating M1 macrophages by calycosin from hairy stamens,one of the main chemical components of Astragalus membranaceus The main results were as follows:(1)the cytotoxicity of calycosin on primary peritoneal macrophages and primary bone marrow macrophages was detected by CCK-8cytotoxicity test and flow cytometry.(2)effect of calycosin from hairy stamens on the production of inflammatory factors in primary macrophages stimulated by inflammation by real-time quantitative PCR.(3)High throughput transcription sequencing was used to find out the possible signaling pathway.Western Blot detection of calycosin in hairy stamens could inhibit the phosphorylation of p65 and reduce the transcriptional activity to reduce inflammation.(4)calycosin can effectively improve the symptom score of collagen-induced arthritis in mice.(5)ABOG,TRAP and Micro-CT showed that calycosin could effectively improve the destruction of bone and cartilage,inhibit synovial hyperplasia and reduce bone loss in collagen-induced arthritis mice.(6)the changes of liver and kidney biochemical indexes(AST,ATL,CRE,BUN)were detected by automatic biochemical analyzer.3.Study on the treatment of collagen-induced arthritis with triptolide nano-dosage form,one of the main chemical components of Tripterygium wilfordii.The main results are as follows:(1)the hydrophobic drug triptolide was encapsulated into the hydrophobic region of nanoparticles by using the hydrophobic effect of the material.(2)p H responsive poly(dimethylaminoethyl methacrylate)(PDMAEMA)was introduced into the hydrophilic part.(3)The characterization of nano-drugs was detected by dynamic light scattering and transmission electron microscopy.(4)the release efficiency of nano-drugs in vitro was measured in different p H environments.(5)CCK-8 and flow cytometry were used to detect the effect of triptolide nanoparticles on apoptosis.(6)the activity of ROS was detected by flow cytometry.(7)Real-time quantitative PCR was used to detect the anti-inflammatory effect of Tripterygium wilfordii nano-drugs at the cell level.(8)the enrichment efficiency of Tripterygium wilfordii nano-drugs in the joints of mice was observed by in vivo imaging of small animals.(9)ABOG,TRAP and Micro-CT were used to observe the effects of different doses of Tripterygium wilfordii nano-drugs on articular cartilage destruction and synovial hyperplasia in mice.(10)the changes of AST,ATL,CRE and BUN in serum of liver and kidney biochemical indexes.(11)HE staining was used to observe the effects of triptolide nanoparticles on liver,kidney and spleen of mice.Results:1.We have developed a prescription of Tripterygium wilfordii combined with astragalus membranaceus in accordance with the concept of "strengthening justice and dispelling evil" in traditional Chinese medicine,which provides ideas for clinicians in the treatment of rheumatoid arthritis.2.We confirmed that calycosin,one of the main chemical components of Astragalus membranaceus,can effectively improve collagen-induced arthritis in mice,and its mechanism may be through regulating M1 macrophages,inhibiting p65 phosphorylation and reducing transcriptional activity,so as to achieve the purpose of anti-inflammation.3.According to the physical and chemical properties of triptolide,one of the main chemical components of Tripterygium wilfordii,the dosage form of triptolide was successfully modified.on the basis of its clear mechanism,the curative effect was increased and the toxicity was effectively reduced by dosage form modification.Conclusion:Calycosin and triptolide can treat collagen-induced arthritis mice.