| Inflammation and pain are the two main symptoms of rheumatoid arthritis (RA),and the clinical therapeutic purposes of RA are anti-inflammatory and analgesic, ourexperiments is according to the technical requirements of the traditional Chinesemedicine drug research, and observing Wutou injection of a variety of animalmodels of inflammation, immune and analgesic effects of its anti-RApharmacodynamic studies, and AA model of its mechanism how to act, which findpharmacological basis for its clinical application.1. Anti-inflammatory experiment1.1Carrageenin induced rats paw swellingWutou injection were given to the rats,on7th day, carrageenin injection wasadministered subcutaneous resulting paw swelling of rats, from time to time thedegree of swelling was examined and we calculated the rate of swelling. The resultsshowed, Wutou injection0.8,1.6mg/kg dose all could significantly inhibitcarrageenin inducing rats paw swelling.1.2Formaldehyde induced rats paw swellingReplication rat paw swelling model induced by formaldehyde, the degree ofswelling was monitored from time to time.Wutou Injection0.8,1.6mg/kg dose couldsignificantly inhibit rat paw swelling induced by formaldehyde from6h to48h.Itindicated that Wutou injection could significantly inhibite rat paw swelling inducedby formaldehyde.1.3The permeability of the blood capillary in abdominal cavity of miceEvan’s blue was administered through intravenous injection, Wutou injection0.5,1.0,2.0mg/kg could not decrease the content of Evan’s blue in abdominal cavity.The results showed Wutou injection had no effect to the permeability of the bloodcapillary in abdominal cavity of mice.1.4Leukocytopenia from the subcutaneous CMC-Na cysts of ratsAfter administration seven days, counting the number of leukocyte in CMC-Nacysts, the results showed, Wutou injection0.8mg/kg dose could significantly inhibitLeukocyte from the CMC-Na cysts of rats at3h of injection. Otherwise, Wutouinjection1.6mg/kg dose could significantly decrease the number of Leukocyte from the CMC-Na cysts of rats at3h and7.5h of injection.1.5Granuloma induced by cotton ball in ratsAfter planting cotton ball in inguinal, Wutou injection were given to the ratsand lasted for seven days.On8th day, dissection of granuloma and calculatinggranuloma index. The results showed, Wutou injection0.8,1.6mg/kg dose couldsignificantly inhibit granuloma index, but thymus index, spleen index and adrenalindex did not change significantly.2. The effect of Wutou injection on immune system2.1Mouse T lymphocyte transformation testMitogen ConA in vitro stimulation of murine T lymphocyte proliferativeresponse was detected by MTT. The results showed that Wutou injection2.0mg/kg dosegroup T lymphocyte stimulation index was significantly reduced, which showed that Wutouinjection high-dose group can inhibit T lymphocyte transformation.2.2Mouse peritoneal macrophages to produce IL-1function testsWutou injection can significantly reduce mouse peritoneal macrophages toproduce the amount of IL-1. IL-1samples diluted1/5, Wutou injection all dosegroup can significantly inhibit the amount of mouse peritoneal macrophages toproduce IL-1. IL-1samples1/10dilution, Wutou injection1.0,2.0mg/kg dose groupcould significantly inhibit murine peritoneal macrophages to produce the amount ofIL-1.3. The effect of analgesic activity3.1Writhing experimentUsing acetic acid-induced mouse writhing, noting the times of mouse writhingand latent period. The results showed, Wutou injection1.0,2.0mg/kg had notableaction on relieving acetic acid-induced mouse pain and reducing writhing times.3.2Hot-plate experimentRecording the points which mice tim hind foot before drug and after drug atdifferent time, to calculate the pain threshold percentage. Wutou injection1.0mg/kgcould significantly improving mice hot plate test pain threshold at4h, and Wutouinjection2.0mg/kg could significantly improve mice thermal plate method of painthreshold at1h,1.5h,2h,3h,4h,5h, Wutou injection0.5mg/kg had no significanteffect on pain threshold of the mouse hot plate.3.3Light-hot experiment Recording the points which rat tail flick time as a pain threshold before drugand after drug at different time, to calculate the pain threshold percentage. Theresults showed that, Wutou injection0.8mg/kg could significantly improve the painthreshold induced by light thermally at1h,2h,3h after drug, Wutou injection1.6mg/kg can significantly increase the pain threshold induced by light thermally at1h,2h,3h,4h after drug, Wutou injection0.4mg/kg caused no significant effect onthe pain threshold induced by light thermally.4. Therapeutic effects of Wutou injection on Adjuvant Arthritis in Rats and itsMechanismOn18th day that the arthritic model of rats was induced by Freund’s adjuvant,Wutou injection (in dosage of0.4,0.8,1.6mg/kg) were given to the rats viaintramuscular administration once a day and lasted for fourteen days. The swellingdegree in both hind paws were examined at different time after treatment and wecalculate the rate of swelling. On19th day after inflammation, PMφ’function wasevaluated, the content of PGE2in inflammatory tissue fluid was measured; thechanges of hemorheology was determined; the contents of TNF-α, IL-1β, IL-6,MDA, NO and SOD in serum were measured in all groups. The results showed theprimarily swelling in right hind paw and the secondum swelling in left hind pawwere relived significantly, the function of PMφ, the content of PGE2ininflammatory tissue fluid, the blood viscosity of10/s,60/s,120/s and erythrocytesedimentation rate were reduced significantly; At the same time, the contents ofMDA, NO, IL-1β and TNF-α in serum were decreased significantly, and activity ofSOD was increased notably, but the content of IL-6in serum had no significanteffect.Wutou injection had anti-inflammatory effect and can inhibit the activity of theimmunity. Wutou injection had certain extent effect to analgesic. Wutou injection hadtherapeutical effect to AA, which may be by inhibiting the activation of PMφ, enhancing therat’s capability of eliminating the free radicals, decreasing the levels of TNF-α, IL-1β andIL-6in serum and improving the changes of hemorheology. This research providesscientific grounds for treating RA. |
PDF Full Text Request