Experimental Study On Detoxicated Of Aconitum Carmichaeli Debx Effect In The Rat Model Of Osteoarthritis

Objective Through preparation to detoxicated of Fuzi and to establish the animal model of osteoarthritis in rats,under the d-fuzi intervention,observation the therapeutic effects of osteoarthritis to d-fuzi,discuss the possible mechanisms in the treatment of osteoarthritis of the d-fuzi.Methods To preparation of the d-fuzi,the alkaloids content of d-fuzi by the HPLC analysis method,and through the QTOF-MS analysis method to measure the chemical composition.In vivo experiment,The forty-five SD rats,aged eight weeks,were randomly divided into three groups:blank groups,model groups,d-fuzi groups.and fifteen rats in each group.No treatments to the blank groups,and others through were injected with iodine acetic acid(1.25g)in articular cavity to establish osteoarthritis animal model.The d-fuzi groups drink the d-fuzi recipe through intragastric administration,Blank control group and model group also through intragastric administration give physiological saline.To give medicine four weeks after the chloral hydrate anesthesia,conduct by Radiographic analysis and Histopathologic analysis to take on both sides of rats knees.In vitro experiment.The ten SD rats,aged four weeks,were randomly divided into two groups:blank groups and d-fuzi groups,and five rats in each group.The d-fuzi groups according to the drug dosage of lOg per kilogram of body weight of rats for conversion to intragastric administration,the blank group with 0.9%physiological saline.one times a day,continuous seven days.At the end of intragastric administration 2h later,anesthesia animals and take blood from heart,through centrifugal to get blank and drug-containing serum.Configured to concentration respectively 5%,10%,15%and 20%of the blank serum and d-fuzi-containing serum.Take the blank group rats knee joint to foster cartilage cell,After identify of toluidine blue,use the third generation of cartilage cells for experiment.Respectively to join the different concentrations of d-fuzi-containing serum(5%,10%,15%,20%),each concentration with 10 duplicate hole;Control group with the blank serum intervention,Use the MTT fresh solution to testing after fostering 24h,48h,72h.Conclusion(1)D-fuzi by HPLC to pharmaceutical analysis,the Baifu table' saconitine,mesa-conitine and hypaconitine significantly reduced by the traditional method of concoct.And with long fried after 120min,mesaconitine and hypaconitine further decrease in the content of alkaloid,including complete decomposition to disdetect aconitine.ByQTOF-MS analysis,there are 23 to d-fuzi of aconite alkaloids component.(2)Radiographic analysis:In the OA group,the knee articular cartilage degeneration was obviously,the joint dissolution,bone hyperplasia,cartilage surface uneven,especially evident in tibia platform.Compared with control group,the tibial bone mineral density decreased significantly(P<0.01).In d-fuzi group,the articular cartilage degeneration is not obvious,articular cartilage surface smooth,the joint space is normal,compared with control group,the tibial bone density has no obvious statistical significance(P>0.05).(3)Histopathologic analysis:In model group,the tibial cartilage were severely damaged,there lack of cartilage,cartilage cell necrosis,osteophyte formation,subchondral bone exposure,and sclerosis of subchondral bone,cyst,trabecular bone area to reduce.Compared with control group,Mankin score significantly higher(P<0.01).In d-fuzi group,reduced the articular cartilage damage,not seen the obvious bone defects,part of reduce the trabecular bone,not see the clearly subchondral bone sclerosis,cysts.Compared with model group,Mankin score significantly reduced(P<0.01).(4)Cartilage cell proliferation:Compared with the blank serum,d-fuzi medicated serum can promote the appreciation of chondrocytes,and depend on the concentration and the,especially in the 10%serum concentration significantly(P<0.01).Also,compared with the blank serum medicated+MIA group,d-fuzi medicated serum+MIA group can obviously reduce the iodine acetic acid damage to the cartilage cells,especially evident in the cultivation of 24h and 48h(P<0.05),It is suggesting to effect of protective cartilage cells of d-fuzi.Results Through preparation process improvement to Baifupian,preparation to d-fuzi,validation reached to no poison effect.Efficacy study found that d-fuzi protective of cartilage,it can promote the appreciation of chondrocytes,reached to no poison but protection effect.