Study On Activity Screening And Mechanism Of Action On RA Of The Extracts From Capparis Spinosa L.

In this experiment,the fruit of Capparis spinosa L.was used as the research material.Through the organic solvent extraction method:petroleum ether,ethyl acetate,n-butanol extract.In this paper,rutin,p-hydroxybenzoic acid and isoquercetin were analyzed by HPLC.At the same time,different concentrations of n-butanol extract from Capparis spinosa L.were used to interfere with MH7A cells,CCK-8 colorimetry was used to detect the proliferation of MH7A cells,the apoptosisof MH7A cells was detected by flow cytometry.In addition,by establishing a bovine type Ⅱ collagen induced arthritis(CIA)rat model,60 SD rats were divided into six groups:High,medium and low dose group of n-butanol extract from Capparis spinosa L.,Blank group,Model group,Positive group(methotrexate).Using protein chip technology to determine RA related inflammatory cytokines in the serum of various groups of rats,including:pro-inflammatory cytokines TNF-α,IFN-y,IL-1β IL-1α,MCP-1,IL-2,IL-6,anti-inflammatory cytokines:IL-13,IL-10,IL-4,and GO enrichment analysis and KEGG signaling pathway enrichment analysis.The main results are as follows:(1)The results of bacteriostasis and antioxidation showed that the extracts of ethyl acetate and n-butanol was better.(2)The results of HPLC analysis showed that rutin,p-hydroxybenzoic acid and isoquercetin had good linear relationship in the range of 0.5～16,1～32,0.5～16 μg/mL(r>0.9990).RSD value of precision,stability and repeatability test were all less than 2.0%.Average recovery rate of sample addition was 84.81%～101.04%.(3)CCK-8 method and flow cytometry were used to detect the proliferation and apoptosis of MH7A cells.The results showed that the n-butanol extract of Capparis spinosa L.had a significant inhibitory effect on the proliferation of MH7A cells and could induce apoptosis.(4)The results of protein microarray showed that the high,medium and low dose of n-butanol extract from Capparis spinosa L.could significantly down regulate the pro-inflammatory cytokines and up regulate the anti-inflammatory cytokines;the enrichment analysis of GO and KEGG showed that the inflammatory cytokines related to RA were mainly enriched in the regulation of NO biosynthesis,NO metabolism,active oxygen biosynthesis process,metabolism of cytokines and chemokines,the metabolic process of chemokines,the production of immune response molecular mediators,the positive regulation of cell secretion,tumor necrosis factor receptor superfamily,IL-2 receptor,IL-6 receptor,IL-1 receptor,JAK/STAT signaling pathway,IL-17 signaling pathway,and the interaction between cytokines and cytokine receptor.