Effect Of Puerarin On Matrix Metalloproteinase And Its Inhibitor Of Cartilage Of Rabbit Knee Joint With Osteoarthritis

Objective:Knee osteoarthritis(KOA) is one of the most clinical common,frequently-occurring disease,with the aging population,the disease is increasingly becoming social and family burden.Articular cartilage degeneration in the disease, destruction,subchondral bone sclerosis as the main pathological features of the degenerative articular cartilage changes in the incidence of osteoarthritis(OA) and the core of the pathological basis.This study on the articular cartilage matrix metalloproteinases and their inhibitors tries to explore the effect of Puerarin(Pue) on OA articular cartilage, as well as its mechanism on treatment of KOA.Method:5 for the experimental group 50 weight 2kg(±200g) healthy adult rabbits(half male and female) animals were randomly divided into normal group,model group,Sodium Chloride group,Hyaluronic Acid group,Puerarin group by 50 healthy adult rabbits(half male and female),each one weights 2kg(±200g).Of 10 randomly selected as the normal group,and the remaining 40 were duplicated by Hulth technique,or cutting the medial collateral ligament,anterior horn of medial meniscus,anterior cruciate ligament model approach.After model surgery 1 week all rabbits were drived for 4 weeks,that is a model of KOA.Osteoarthritis model to be established,did not do anything to deal with the normal group;the model group only by Hulth model and non-drug treatment;Sodium Chloride group,Hyaluronic Acid(HA) group,Pue group in the first 1 week after preparation of animal models were put into strict aseptic operation,were given the knee joint injection of 0.9%Sodium Chloride,HA,Pue each 0.3ml,2 times per week,that is,5 weeks continuously.In 1 week after end of treatment,all were sacrificed by air embolism,and the knee joint cavity was exposed.①To observate,photograph and survey the morphological and histological changes of rabbits knees articular cartilage cartilage with eyes and microscope.②To detect the expression of MMP-1 and TIMP-1 by immunohistochemistry.③To survey the mRNA expression of MMP-1 and TIMP-1 by RT-PCR.Results:①Smooth articular surface shiny,transparent and no obvious defect in the cartilage and the new biological material,without synovial hyperemia were found by naked eye in normal group.Articular surface of the model is not smooth, matte,opaque cartilage there is an obvious defect and the new biology,synovial swelling congestive dark color.Articular cavity Pue group was no significant new biological,dark color, cartilage worse than the smooth but transparent.Sodium Chloride group was similar to the model group;HA group was similar to groups.Pue group is more formation of cartilage cells,cartilage cells,necrotic degeneration of the surface than the model group and the Sodium Chloride group with light rare exudation of inflammatory cells by microscope,we could see new blood vessels and fibroblasts.②2The expression rates of MMP-1 and TIMP-1:there was a little MMP-1 and TIMP-1 positive expression in normal group.In model group,MMP-1 positive cell count was significantly increased than the normal group,and its expression of TIMP-1 was negative.Sodium Chloride group was similar to model group.MMP-1 positive cells of Pue group were lower than the model group,decreased expression than the model group,HA group is similar to Pue group.TIMP-1 in the puerarin group and the expression of hyaluronic acid group were lower,and no significant differences.③The Positive expression rates of MMP-1 and TIMP-1 mRNA:the mRNA expression rates of MMP-1 and TIMP-1 in knee cartilage of nomal groups were very low.The mRNA expression rates of MMP-1 and TIMP-1 in knee cartilage of model group and significantly increased than nomal group(P<0.05).The expression of TIMP-1 was negative in model group.The mRNA expression rates of MMP-1 of Pue and HA group were higher than nomal group and lower than model group.The mRNA expression rate Of TIMP-1 Of the group of the Pue was significantly different than model the and Sodium Chloride group(P<0.05).The mRNA expresion rates of TIMP-1 in Pue group was very low and no significant difference(P>0.05).Conclusions:Matrix metalloproteinase is able to reflect changes in bone metabolism of articular cartilage,its articular cartilage extracellular matrix degradation have to increase the damage to cartilage degeneration,that has a very important relationship.The study found,through experimental animal model of cartilage tissue of MMP-1 expression was significantly increased,and TIMP-1 expression in very few,and the cartilage extracellular matrix of collagen typeⅡcontent decreased.After the treatment of animals cartilage MMP-1 expression decreased in Pue group,while TIMP-1 expression increased on articular cartilage,the extracellular matrix increases collagen content.That is,Puerarin could put off extracellular matrix collagen degradation with a certain degree, and may slow down the course of osteoarthritis progress.The study supply theory foundation for Puerarin on OA treatment.