| ObjectiveTo study the mechanism of the active ingredient of Aconitum flavum Hand.-Mazz oncure RA.Methods1. We extracted Aconitum flavum Hand.-Mazz with Solvent method, enriched theeffective parts of Aconitum flavum Hand.-Mazz by Macroporous Absorption Resin and CationExchange Resin, determined the content of the total flavonoids in effective parts by UVspectrophotometry.2. Adopt MTT assay to determine and filtrate the effective parts and active ingredients ofAconitum flavum Hand.-Mazz.3. Male SD rats of72, weight180±20g, randomized divided into9groups. In additionto the blank group, the rest groups of the rats toe were intradermal injected in Freund’sadjuvant to prepare rat adjuvant arthritis (adjuvant arthritis, AA) model. After modeling8days later，gavaged the rats (ig,dosing volume of1.0ml/100g).30%alcohol active extracts ofhigh, medium and low group to30%alcohol parts of the solution (60,30,15mg/kg) orally;Gavaged rats the non-alkaloid fraction solution (120,60,30mg/kg) of non-alkaloid fraction ofhigh, medium and low group. Control group was gavaged with (9.45mg/kg) TG tabletssuspension; normal group and model group was gavaged with0.5%CMC-Na solution.Continuously gavaged to the rats21days, then the rats were accordingly treated2hours laterin the last gavagation. 4. Adopt HE staining, joint swelling, polyarthritis index, weight and other indicat-ors to evaluate the AA model of the rats, and use these indicators to observed thetherapeutical effect of effective parts of Aconitum flavum Hand.-Mazzto AA rats.5. Adopt ELISA assay to investigate IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-17and thecontent of TNF-alpha of serum of AA rats(p<0.05); Adpot immunohistochemical method(Immunohistochemisty,IHC) to observe vascular endothelial growth factor (vascularendothelial growth factor,VEGF) and the expression of matrix metalloproteinase-3(matrixmetalloproteinases-3, MMP-3) in synovial tissue.Results1. We got0.3kg extractum liquidum from3kg original materials, obtained12.3g from30%alcohol by Macroporous Absorption Resin,yield of4.1%; we got4g non-alkaloidfraction by Cation Exchange Resin, yield of50%.2. The total flavonoids content was1.2%in the raw herb of Aconitum flavumHand.-Mazz. The total flavonoids content was9.6%in30%alcohol by Macroporous AbsorptionResin by Cation Exchange Resin, non-alkaloid effective part of total flavonoids was29.26%.3. Linarin(0.05,0.01mg/kg),30%ethanol elution parts by Macroporous resin (3.12g/kg)can significantly inhibit the ConA-induced of mice with spleen lymphocytesproliferativeresponse.4. The dose group (60mg/kg) non-alkaloid fraction of30%ethanol elution parts (60,30mg/kg) by Macroporous resin has anti-inflammatory and analgesic effects, can reduce thesecondary paw swelling in AA rats（p<0.05）, and significantly reduce rat polyarthritisindexand ease the decline weight of AA rats（p<0.05）, improve the pressure pain tolerancevalue of AA rats（p<0.05）.5. The non-alkaloid fraction dose group (60mg/kg) of the30%ethanol elution parts (60,30mg/kg) play anti-inflammatory and immunomodulatory effects through inhib iting the AA rats inflammatory cytokines IL-1beta,IL-4, IL-5,IL-6, TNF-α secretion andincreasingIL-2, IL-17.6. The non-alkaloid fraction dose group (60mg/kg) of the30%ethanol elution parts(60,30mg/kg) can prevent erosion of the cartilage and subchondral bone by down-regulatingVEGF expression in the vascular wall, and reduce the proliferation of endothelial cells,vascular exclusion newborn and the expression of vascular endothelial cells MMP-3.Conclusion1. The active parts of Aconitum flavum Hand.-Mazz. have pharmacological effects oftreatment of main ingredients may be flavonoid composition.2. The active parts of Aconitum flavum Hand.-Mazz. have pharmacological effects ofimmunoregulation.3. The therapeutical effect of effective parts of Aconitum flavum Hand.-Mazz to RA, itmaybe relate to the production of the cytokines, reduce the release of inflammation of themeson and regulate the immune function and subchondral bone by down-regulating VEGFexpression in the vascular wall, and reduce the proliferation of endothelial cells, vascularexclusion newborn and the expression of vascular endothelial cells MMP-3. |
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