| BackgroundOsteoarthritis(OA)is the most common degenerative joint disease that affects millions of middle-aged and elderly people worldwide and is characterized by joint pain and chronic inflammation.In clinical treatment of arthritis,analgesic and anti-inflammatory drugs are routinely used,which have large side effects.Joint synovial inflammatory factors are mainly derived from synovial cells.The main functional cells of synovial tissue are Fibroblast-like synoviocytes(FLS),but the mechanism of FLS in OA and other inflammatory joint diseases is still unclear.It is not fully understood,and there is an urgent need to treat joint synovial inflammation by drug research intervening in FLS.Ski is the intracellular homolog protein of avian oncogene v-ski,which also inhibits wound inflammation.We evaluated the potential of the endogenous transcriptional regulator Ski as an anti-inflammatory alternative for OA through experimental studies in animal models and in vitro cells.ObjectiveThe OA prototype model was induced by Monosodium iodoacetate(MIA),and the primary culture method of FLS and the in vitro inflammatory model were established to explore the anti-inflammatory effect of endogenous factor Ski on FLS in knee osteoarthritis(KOA)rats.MethodsThe KOA of SD rats was induced by MIA,and the synovial tissue of knee osteoarthritis was isolated under sterile conditions for primary culture of synovial tissue and LPS-induced secretion of primary FLS pro-inflammatory factors to establish an in vitro FLS inflammation model.In vitro,Ad-ski was transfected to overexpress Ski,and it was observed whether Ski could inhibit the expression of pro-inflammatory factors IL-1β,IL-6 and TNF-α mRNA and protein in FLS and U-937 cells under LPS stimulation.In addition,it was also observed whether Ski could up-regulate the protein expression of the anti-inflammatory factor IL-4 and down-regulate the protein expression of downstream molecules related to the NF-κB inflammatory signaling pathway,so as to explore the attenuating or inhibiting effect of Ski on OA-induced synovial inflammation.Results1 After MIA was injected into the joint cavity of rats,it destroyed chondrocytes and glycolytic energy metabolism,the expression of HIF-1α continued to increase,the secretion of IL-1β and TNF-α increased,the weight-bearing ability of hindlimbs of rats decreased,and the osteoarthritis biomarker osteopontin was overexpressed,and KOA modeling was successful.2 The synovial tissue of the rats in the Sham group and the control group(KOA)group was separated,and the primary cultured synovial cells from the 3rd to 7th passages under physiological and pathological conditions were obtained by collagenase type I digestion,the FLS purity is above 98%,and it has the biological activity of synovial tissue FLS.Primary cultured FLS can be used as a cellular model for the study of OA in vitro.3 The in vitro inflammatory model of FLS was successfully induced by LPS,and the optimal concentration of LPS to stimulate FLS was 1 μg/mL,and the optimal duration was 3 hours.4 It was found that Ski had an anti-inflammatory effect on KOA synovial FLS and macrophages,and down-regulated the proteins of NF-κB p65-related downstream factors VEGF,VCAM1 and MMP-(1-3),significantly decreased the expression of pro-inflammatory factor IL-6 stimulated by LPS and significantly increased the expression of anti-inflammatory factor IL-4.4.1 After FLS was transfected with Ad-ski,it could not only reduce the basal expression of IL-1β and TNF-α protein in FLS,but also blocked the high expression of IL-1β and TNF-αinduced by LPS.The effect on the protein expression of IL-1β and TNF-α was the same as that of dexamethasone.4.2 After U937 cells were transfected with Ad-ski,it could not only reduce the basal expression of IL-1β and TNF-α protein in U937 cells,but also blocked the high expression of IL-1β and TNF-α induced by LPS.The effect on the expression of IL-1β and TNF-α protein was the same as that of dexamethasone.4.3 After Ad-ski transfection,the proteins of the downstream factors VEGF,VCAM1 and MMP-(1-3)related to NF-κB p65 were changed,indicating that the anti-inflammatory mechanism of Ski may be related to the NF-κB signaling pathway.ConclusionsSki has an inhibitory effect on synovial cell inflammation of SD rat with knee osteoarthritis induced by monosodium iodoacetate.If Ski is an endogenous anti-inflammatory factor,it has less immunogenicity and less possible side effects.The development of Ski as a drug for the treatment of KOA should have great potential.Of course,to explore whether the anti-inflammatory effect of Ski is through the NF-κB signaling pathway,the next step is to complete the mechanism study;and then expand from cell models to animal models to clinical applications. |
PDF Full Text Request