An Experimental Study Of Achyranthes Bidentata Polysaccharide On The Proliferation And Apoptosis Of Articular Chondrocytes In Degenerative Human Knee Osteoarthritis

BackgroundKnee osteoarthritis has become a major chronic degenerative disease with very high disability.The use of traditional Chinese medicine to prevent and treat this disease is a major demand for human health.Knee Osteoarthritis(KOA)is a clinical manifestation of articular cartilage degeneration and pain,stiffness and dysfunction.It consumes a large number of residents' health protection resources and becomes a huge achievement for our healthy China 2030 strategy.challenge.At present,its pathogenesis is still not fully understood.From a pathological point of view,it is a chronic progressive osteoarthrosis characterized by articular cartilage degeneration and bone hyperplasia around the joint.After more than 30 years of clinical experience and literature research,the instructor group believes that the occurrence and development of knee osteoarthritis involves many factors,and "virtual,phlegm and poison" are closely related to this disease.Middle-aged and old people with qi and blood disharmony,spleen and stomach dysfunction,liver and kidney deficiency,the imaginary first,meridian obstruction,blood stasis endogenous,long-term illness must be phlegm,re-feeling cold,cold and external evil,evil victory is the poison,It turns out to be poisonous,and then "smoke,poison" to become imaginary,which leads to the "sin,sputum,and poison three evils".The instructor's group has added the flavored live-active parasitic mixture as one of the main treatments for KOA prevention and treatment.It has been clinically carried out for many years,and through clinical observation,it has been confirmed that the addition of the unique living parasitic mixture KOA can relieve local pain symptoms,effectively improve knee joint function,and reduce joint fluid.Inflammatory factors.Flavored solitary parasitic soup composition: solo,mulberry,Eucommia,Achyranthes,Astragalus,Gentiana,Poria,Cinnamon,Wind,Chuanxiong,Codonopsis,Licorice,Angelica,Angelica,Rehmannia,South Star,Astragalus,Papaya,Wei Lingxian.We have used self-medicated and flavored live-active parasitic mixture to replenish liver and kidney,promote blood circulation to remove blood stasis,detoxify and remove phlegm to treat knee osteoarthritis,and achieved good clinical results.The achyranthes is one of the main ingredients of the flavored live parasitic mixture.This experiment focused on the effects of Achyranthes bidentata polysaccharides on proliferation and apoptosis of human knee joint chondrocytes,and laid a theoretical foundation for the treatment of knee osteoarthritis with Achyranthes.Purpose1.Verify whether Achyranthes bidentata polysaccharide has toxic effects on chondrocytes;2.Confirmed the effect of Achyranthes bidentata polysaccharide on chondrocyte proliferation and apoptosis;3.To determine the effect of Achyranthes bidentata polysaccharides on the expression of Caspase-3/9 protein in chondrocytes,and to explore the theoretical mechanism of Achyranthes bidentata in the treatment of knee osteoarthritis at the cellular level.Method1.Primary human knee chondrocytes were obtained and cultured by trypsin and type II collagenase two-step enzymatic digestion method,and identified by inverted microscope and toluidine blue staining;2.Using IL-1? to model the chondrocyte environment of human knee osteoarthritis,and intervene with different concentrations of Achyranthes bidentata polysaccharides;3.CCK-8 method was used to detect the toxic effect and the value-added effect of Achyranthes bidentata polysaccharide on human knee joint chondrocytes;4.Flow cytometry was used to detect the effect of Achyranthes bidentata polysaccharides on apoptosis of human knee articular chondrocytes.5.Western blot was used to detect the effect of Achyranthes bidentata polysaccharide on the expression of apoptosis protein Caspase-3/9 in human knee articular chondrocytes.Result1.The primary human knee cartilage cells were successfully obtained and cultured by trypsin and type II collagenase two-step enzymatic digestion method,and after identification,meet the experimental requirements;2.The apoptotic rate of chondrocytes increased significantly after IL-1? intervention,meeting the requirements of chondrocyte model of knee osteoarthritis;3.The CCK-8 assay showed that the Achyranthes bidentata polysaccharide had no toxic effect on chondrocytes at a certain concentration,P>0.05.Compared with the control group,the concentration of Achyranthes bidentata polysaccharides at 50mg/L,100mg/L and 200mg/L could promote chondrocyte proliferation,P<0.05;4.The results of flow cytometry showed that the apoptosis rate of chondrocytes in 50mg/L,100mg/L and 200mg/L Achyranthes bidentata polysaccharides group decreased significantly compared with the control group,P<0.05.5.The results of Western blot showed that the expression level of apoptosis protein Caspase-3/9 in chondrocytes of experimental group was significantly decreased compared with the control group,P<0.05.ConclusionA certain concentration of Achyranthes bidentata polysaccharide can promote the proliferation of chondrocytes and inhibit the apoptosis of chondrocytes,thereby maintaining the stability of cartilage matrix and delaying the degeneration of chondrocytes.The mechanism may be through inhibition of Caspase-3/9-CASP3 pathway.