Xuelian Injection Prevention And Treatment Of Osteoarthritis And Its Mechanism

[Objective]OA(osteoarthritis)of knee joint is a kind of common disease.It effects seriously on work and live of people.It is thought that the pathogenesis of osteoarthritis of knee joint results from such factors as stress,cytokine(CK), apoptosis,auto-immune,heredity,inflammation,matrixmetallo proteinase(MMP)and tissue-inhibitor mentallo proteinase(TIMP).However,the pathogenesis of OA is still not dearly determined.At present,the authors with an attempt to further explore the mechanism of OA.The pathology of OA includes mainly the histology and metabolic change of the chondrocyte and the biochemical and configuration change of extra cellular matrix (ECM)and secondary synovitis.The well-balanced chondrocyte can spontaneously excrete microcrystalline interlukin-1β,but the chondrocyte of knee joint osteoarthritis can excrete interlukin-1βexceptionally manifold,and consequently bring on apoptosis and decreasing level of collagenⅡand PG(proteoglycan).Finally extra cellular matrix(ECM)suffer degradation.The mail remedy of knee joint osteoarthritis is to stave off the cataplasia of cartilage and to promote the regenesis of hyaline cartilage.As an important dispelling rheumatism medicine,XueLian injection has been widely used in curing RA (rheumatoid arthritis)and OA(osteoarthritis)and has gained well curative effect.This topic tries to find out the mechanism and the preventive method of XueLian injection.We infused XueLian injection into osteoarthritis rabbit model by intramuscular injection to study the effect on histology,collagen phenotype,ultramicrostructure,interlukin-1βlevel,HA,NO and OFR in OA model.[Method]OA model animals were randomly divided into 4 groups:group A,group B,group C and group D.Animals in group A received XueLian injection and animals in group B received sterilized normal saline intramuscularly,group C was control and received nothing and another group(group D)was used for normal control.After the animals were killed,cartilage specimen were obtained and sections were made and stained With HE/SOFG for Mankin score;change of ultramicrostructure of cartilage was observed with transmission electron microscope. After the animals were killed,the IL-1βlevel in joint fluid of A group was determined and compared with that of group B,C,D.in-vitro:cartilage of rabbit were obtained, isolated and cultured.The secondary passes cells were divided into groupⅠandⅡ, different concentrations of XueLian injection were added to groupⅠfor 72 hours; groupⅡa received different concentrations of XueLian injection for 24 hours and then received LPS for 72 hours,groupⅡb received LPS first,after 24 hours different concentrations of XueLian injection solution were added.72 hours later,the supernatant of each group were collected and the IL-1βlevel was determined by RIA. After the animals were killed,cartilage specimen were obtained and sections were tested by immunohistochemistry.The hyaluronic acid level in joint fluid of A group was determined and compared with that of group B,C,D.The nitric oxide level in osteoarthritis serum and superoxide dismutase and malondialdehyde level in osteoarthritis synovialis of A group was determined and compared with that of group B,C,D.[Result]Mankin score in group A was better than that in group B and C at each test period;cartilage impairment degree in group A was lower than in group B and C,and impairment manifestation in group B and C was similar on the whole. XueLian injection can significantly decrease the interlukin-1βlevel in the joint fluid of OA model animal and the in-vitro cultured chondrocyte supernatant.Rresult of IHC shows that positive stain area of collagenⅠWas lower in group A than that in group B and C.and positive stain area of collagenⅡWas higher than that in group B and C at each test period;cartilage impairm ent degree in group A Was lower than in group B and C,and impairment manifestation in group D and K was similar on the whole.XueLian injection can significantly increase the hyaluronic acid level in the joint fluid of OA model animal.XueLian injection can significantly decrease the nitric oxide level in osteoarthritis serum of OA model animal.[Conclusion]XueLian injection may protect articular cartilage by maintaining and promoting the chondrocyte synthesizing proteoglycan,by decreasing the IL-1βlevel in joint fluid and inhibiting the secretion of IL-1βfrom chondrocytes,by maintaining and promoting the typeⅡcollagen,by increasing the hyaluronic acid level in joint fluid,by decreasing the nitric oxide level in osteoarthritis serum of OA model animal.The first section:Effect of XueLian injection on histomorphology in cartilage of osteoarthritis animal model[Objective]The object of this research is to study the effect of XueLian injection on histology and ultramicrostructurc in cartilage of osteoarthritis animal model[Method]OA model animals were randomly divided into 4 groups:group A,group B,group C and group D.Animals in group A received XueLian injection and animals in group B received sterilized normal saline intramuscularly,group C was control and received nothing and another group(group D)was used for normal control.After the animals were killed,cartilage specimen were obtained and sections were made and stained with HE/SOFG for Mankin score;change of ultramicrostructure of cartilage was observed with transmission electron microscope.[Result]Mankin score in group A was better than that in group B and C at each test period;cartilage impairment degree in group A was lower than in group B and C,and impairment manifestation in group B and C was similar on the whole. [Conclusion]XueLian injection may protect articular cartilage by maintaining and promoting the chondrocyte synthesizing proteoglycan.The second section:Effect of XueLiau injection on interlukin-1 level in osteoarthritis joint fluid and chondrocyte culture supernatant[Objective]The object of this research is to study the effect of XueLian injection on interlukin-1βlevel in OA model and in-vitro culture chondrocyte supematant.[Methods]in-vivo:OA model animals were randomly divided into 4 groups:A XueLian injection group,B Saline group,C Control group. Group A B received XueLian injection im and sterilized normal saline im respectively, group C received nothing,and anothergroup(group D)was used for normal control. After the animals were killed,the IL-1βlevel in joint fluid of A group was determined and compared with that of group B,C,D.in-vitro:cartilage of rabbit were obtained, isolated and cultured.The secondary passes cells were divided into groupⅠandⅡ, different concentrations of XueLian injection were added to groupⅠfor 72 hours; groupⅡa received different concentrations of XueLian injection for 24 hours and then received LPS for 72 hours,groupⅡb received LPS first,after 24 hours different concentrations of XueLian injection solution were added.72 hours later,the supernatant of each group were collected and the IL-1βlevel was determined by RIA. [Results]XueLian injection can significantly decrease the interlukin-1βlevel in the joint fluid of OA model animal and the in-vitro cultured chondrocyte supernatant. [Conclusion]The therapeutic effect of XueLian injection on OA can be achieved by decreasing the IL-1βlevel in joint fluid and inhibiting the secretion of IL-1βfrom chondrocytes.The third section:Effect of XueLian injection on histomorphology in cartilage of osteoarthritis animal model[Objective]The object of this research is to study the efect of XueLian injection on collagen phenotype in cartilage of osteoarthritis animal model.[Method]OA model animals were randomly divided into the expression level of collagenⅠandⅡin mamx was 4 groups:group A,group B,group C and group D.Animals in group A received XueLian injection and animals in group B received sterilized normal saline intramuscularly,group C was control and received nothing and another group(group D)was used for normal control.After the animals were killed,cartilage specimen were obtained and sections were tested by immunohistochemistry.[Result]Rresult of IHC shows that positive stain area of collagenⅠWas lower in group A than that in group B and C.and positive stain area of collagenⅡWas higher than that in group B and C at each test period;cartilage impairment degree in group A Was lower than in group B and C,and impairment manifestation in group D and K was similar on the whole.[Conclusion]XueLian injection may protect articular cartilage by maintaining and promoting the typeⅡcollagen.The fourth section:Effect of XueLian injection on Hyaluronic acid level in osteoarthritis joint fluid[Objective]The object of this research is to study the effect of XueLian injection on hyaluronic acid level in OA model.[Methods]OA model animals were randomly divided into 4 groups:A XueLian injection group,B Saline group,C Control group.Group A B received XueLian injection im and sterilized normal saline im respectively,group C received nothing,and anothergroup(group D) was used for normal control.After the animals were killed,the hyaluronic acid level in joint fluid of A group was determined and compared with that of group B,C,D. [Results]XueLian injection can significantly increase the hyaluronic acid level in the joint fluid of OA model animal.[Conclusion]The therapeutic effect of XueLian injection on OA can be achieved by increasing the hyaluronic acid level in joint fluid.The fifth section:Effect of XueLian injection on Nitric oxide level in osteoarthritis serum and Superoxide dismutase and Malondiaidehyde level in osteoarthritis synovialis[Objective]The object of this research is to study the effect of XueLian injection on nitric oxide level in osteoarthritis serum and superoxide dismutase and malondialdehyde level in osteoarthritis synovialis[Methods]OA model animals were randomly divided into 4 groups:A XueLian injection group,B Saline group,C Control group.Group A B received XueLian injection im and sterilized normal saline im respectively,group C received nothing,and anothergroup(group D)was used for normal control.After the animals were killed, the nitric oxide level in osteoarthritis serum and superoxide dismutase and malondialdehyde level in osteoarthritis synovialis of A group was determined and compared with that of group B,C,D.[Results]XueLian injection can significantly decrease the nitric oxide level in osteoarthritis serum of OA model animal [Conclusion]The therapeutic effect of XueLian injection on OA can be achieved by decreasing the nitric oxide level in osteoarthritis serum of OA model animal...