Effects Of Panax Notoginseng Saponins (PNS) On The Proliferation And Metabolism Of Rabbit Articular Chondrocytes Induced By TNF-α In Vitro

Objective:This study aimed at the effects of Panax Notoginseng Saponins on the proliferation of rabbit articular chondrocytes which are normal and induced by TNF-αcultured in vitro.To observe the the effects of Panax Notoginseng Saponins on the metabolism of proteoglycan(PG) and collagen,and the SOD,NOS of chondrocytes.To probe the possible mechanism of prevention and cure OA with PNS on the cellular level,and providing guidance for clinical practice.Methods:Chondrocytes were isolated from rabbit article cartilage and cultured in vitro.To observe the morphological changes and growth feature by intertrd microscope.The proliferation rates of chondrocytes which were normal and induced by TNF-αwere detected by MTT chromometry,after cultured in different concentrations of PNS(25μg/ml,50μg/ml,100μg/ml,200μg/ml,400μg/ml,800μg/ml).the production of collagen and proteoglycan were estimated respectively by determination of the hydroxyp roline content and Alician Blue method.And the content of SOD and NOS in supernate are tested.Results:The original generation of chondrocytes were in polygonal shape,adhering to the wall after cultured 48 hours.And growed faster when passed to the next generation.The cells maintained the morphology staining pattern within the first three generation,while after the fifth generation,it's changed obviously.Compared with the control group,50μg/ml,100μg/ml,200μg/ml of PNS group could promote the proliferation of normal chondrocytes with remarkable difference(P＜0.05),the 100μg/ml group had the greatest function among them.Compared with the blank control group and the model group,all the 50μg/ml, 100μg/ml,200μg/ml of PNS group could promote the proliferation of chondrocytes which were induced by TNF-αand PNS,and had remarkable difference(P＜0.05).No matter cultured by TNF-αfor 4 hours first and then added PNS or on the contrary,the value of OD had no remarkable difference(P＞0.05).The 100μg/ml of PNS group could promote the anabolism of proteoglycan obviously(P＜0.05),compared with the blank control group.While 50μg/ml, 100μg/ml of PNS group were also promote the anabolism of proteoglycan(compared with the model group).Compared with the blank control group 25μg/ml,50μg/ml,100μg/ml of PNS group could increase the content of collagen remarkabley(P＜0.05).With the concentration of PNS increasing,the function enhanced gradually,while the 400μg/ml,800μg/ml of PNS group were reduced obviously(P＜0.05).Compared with the model group,25μg/ml,50μg/ml,100μg/ml, 200μg/ml of PNS group could increase the content of collagen remarkabley (P＜0.05),and with the concentration increasing,the function enhanced gradually.When the concentration of PNS were higher than 200μg/ml,the content of collagen was reduced,the higher the less.And in the 800μg/ml of PNS group,the content of collagen was remarkably reduced(P＜0.05).At a certain concentration of PNS(during 25μg/ml to 200μg/ml),the chondrocytes' SOD activity was much higher than both the blank control group and the model group(P＜0.05).While the concentration as hingh as 800μg/ml,it's not affected.PNS could reduce the chondrocytes' NOS activity remarkably which were induced by TNF-α(P＜0.05),when the concentration is lower than 800μg/ml.During 25μg/ml to 200μg/ml,PNS had no remarkabkle influence to the NOS activity (P＞0.05),compared with the control group.As the concentration increasing,NOS activity enhanced and much higher than the blank control group(P＜0.05). Conclusions:The Chondrocytes from the second to the fourth generation were perfect celles for study.A certain concentration of PNS could promote the proliferation,promote the matrix synthesis,enhance the SOD activity,reduce the NOS activity of chondrocytes.PNS could counteract the damage of cartilage induced by TNF-α, accelerate the repairability of chondrocytes.So the study of PNS is a momentous and effective way to prevent and cure OA with drugs in clinic practice.