| Objective:(1) To observe changes of several inflammatory cytokines(IL-16,MMP-1) and transcription factors(NF-κB) in injured articular cartilage tissues, and investigate pathogenesis of post-traumatic osteoarthritis.(2) To investigate rhein and diacerein's depressant effects to the expressions of inflammatory cytokines in synovia and serum, and offer these mechanism of action to treat post-traumatic osteoarthritis in clinic.Methods:(1)The rabbit models of injured articular cartilage made by impacting of low(0.9J) and high(6.3J) energy were separated into low energy injured group(L group) and high energy injured group(H group).(2) The chondrocytes of knee joints from normal and the modals were isolated by enzyme digestion and cultured in vitro, then their abilities of survive, proteoglycan synthesis, type-Ⅱcollagen synthesis,mRNA expressions of IL-βand NF-κB, IL-1βsynthesis and MMP-1 synthesis were measured. After the chondrocytes isolated from H group were administrated with Rhein,HA and N-Ac respectively, The changes of the chondrocytes' biologic and pathologic functions were measured to determine drugs' therapeutic action to injured chondrocytes.(3) The type-B synoviocytes of knee joints from normal and the models were got by tissue cultivation in vitro, then their abilities of IL-1βand MMP-1 synthesis,mRNA expressions of IL-βand NF-κB were measured. After the type-B synoviocytes were administrated with Rhein,HA and N-Ac separately,the changes of these functions were measured to determine drugs' therapeutic action to injured synoviocytes.(4) The articular cartilage were viewed by HE stain, safranin O stain to determine its cataplasia degree in different periods, the expression of IL-1β\NF-κB in articular cartilage and synovium were measured by immunohistochemistry stain and the concentrations of IL-1β\MMP-1 were measured by ELISA. Then the relationships between the inflammatory cytokines expressions and cartilage cataplasia were analysised.(5)The rabbits model of H group were given diacerein, celecoxib, HA, and N-Ac separately. Then the articular cartilages and synoviums were measured by HE stain and safranin 0 stain, and the expressions of IL-1βand NF-κB in the articular cartilages and synoviums were measured by immunohistochemistry stain, Further more, the levels of IL-1βand MMP-1 were tested by ELISA. Through above tests, the drugs' effect in the expressions of inflammatory cytokines to injured articular cartilage and tissue cataplasis were investigated.Results:(1)The chondrocytes of H group and L group compared with normal group had less survival rate, and primary cells had lower abilities of adherence and growth. In the chondrocytes, the intensity of in toluidine blue stain and type-Ⅱcollagen IHC was lower than normal's. Expressions of IL-1βand NF-κB in the chondrocyte and concentrations of IL-1βand MMP-1 in cell culture fluid rised(H group>L group>Normal group, P<0.05).(2)In H group, after the chondrocytes were administrated with Rhein, HA, and N-Ac respectively in different concentration, proteoglycan and type-Ⅱcollagen synthesis in them rised,expressions of IL-1βand NF-κB and concentrations of IL-1βand MMP-1 in cell culture fluid fell.The changes were more obvious when Rhein was administrated as 20ug/ml and HA as 4μg/ml.(3) In the type-B synoviocytes of H and L groups, expressions of IL-1βand NF-κB' mRNA and synthesis of IL-1βand MMP-1 significantly rised, but after intervention of three drugs they all fell. The changes were more obvious when Rhein was administrated as 20ug/ml and HA as 4μg/ml.(4)In early stage of H and L group, the concentration of IL-1βand MMP-1 in synovial fluid and serum significantly rised compared with contrast group(P<0.01). The concentration of IL-1βand MMP-1 recoverd in L group after 4 weeks, but maintained at a higher level in H group.(5)In H and L groups, expressions of IL-1βand NF-κB of cartilage and synovium rised. In L group, the expressions concentrated in surface layer of articular cartilage and liner layer of synovium, and were as similar as contrast group after 4 weeks(P>0.05). In H group, the expressions laid in whole surface layer of articular cartilage and liner layer of synovium, and graduately rised as time.The changes were significantly different compared with L and contrast group(P<0.01).(6)In H group administrated respectively with Rhein, celecoxib, HA, and N-Ac, the concentration of IL-1βand MMP-1 in synovial fluid and serum and the expressions of IL-1βand NF-kB fell. Celecoxib, HA, N-Ac had better effect at early stage of treatment, but Rhein had the best effect and alendronate had the least one after 4 weeks.Conclusion:(1)Impacting directly articular cartilage results in some influences: chondrocyte's abilities of adherence and growth fall, and in the chondrocyte and synoviocyte expressions of IL-1βand NF-κB and concentrations of IL-1βand MMP-1 rised.The influences positively correlate with the size of impacting energy.(2) Rhein improves injuried chondrocyte's abilities in growth, multiplication, and synthesis of proteoglycan and type-Ⅱcollagen. Rhein's effects despond on it's dose.In H group, when Rhein is administrated as 20 ug/ml, injuried chondrocyte's abilities in synthesis of proteoglycan and type-Ⅱcollagen are similar as L group's.Rhein reduces the abilities of injuried chondrocyte and synoviocyte in expressions of IL-1βand NF-κB and synthesis of IL-1βand MMP-1.(3) In L group expressions of IL-1βand NF-κB in articular cartilage and synoviocyte increase especially in surface of tissue, and this change lasts 4 weeks after impacting. Change of IL-1βand MMP-1 synthesis in synovial fluid like above, but the increase of IL-1βand MMP-1 synthesis in serum only lasts 2 weeks after impacting.(4)In H group, expressions of IL-1βand NF-κB in articular cartilage and synoviocyte and concentration of IL-1βand MMP-1 in synovial fluid and serum increase and maintain at a high level. The changes show injuried cartilaginous tissue and chondrocyte can't repair automatically, and get worse continuously.(5) In H group diacerein, celecoxib, HA, N-Ac and alendronate can reduce significantly expressions of IL-1βand NF-κB of cartilage and synovium and IL-1βand MMP-1 synthesis in serum and synovial fluid. They inhibit intraarticular inflammation and depress cataplasis of articular cartilage. In early stage of injury,Celecoxib, HA, N-Ac had better effect,but diacerein had the best effect after 4 weeks.
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