Effect of Phlorizin on Osteoarthritis Induced by Anterior Cruciate Ligament Transection in Rats
Randomly divide rats into sham surgery(Sham)group,model group,low-dose Phlorizin(PHL)(PHL-L)group medium-dose PHL(PHL-M)group,high-dose PHL(PHL-H)group,and PHL-H+NF-κB pathway activator PMA(PHL-H+PMA)group.Except for the Sham group,a rat model of osteoarthritis induced by anterior cruciate ligament transection was constructed.Observe the pathological status of rat articular cartilage tissue and evaluate the degree of articular cartilage injury in rats.The Mankin's score was applied to assess the degree of joint cartilage injury.ELISA method was applied to detect the expression levels of TNF-α,IL-6,and IL-1β in joint lavage fluid.The reagent kits were applied to detect the levels of oxidative stress-related indicators such as superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),and malondialdehyde(MDA)in joint lavage fluid.Western blot method was applied to detect the expression levels of NF-κB p65/NLRP3 signaling pathway related proteins,and type Ⅱ collagen(COL-Ⅱ)and matrix degrading enzyme 13(MMP13)in articular cartilage tissue.To investigate the effect of phlorizin(PHL)on anterior cruciate ligament transection induced osteoarthritis in rats by regulating the nuclear transcription factor kappa-B p65/Nod-like receptor protein 3(NF-κB p65/NLRP3)signaling pathway.The results show that,compared with the Sham group,the joint cartilage tissue of rats in the Model group showed obvious pathological damage,with the disappearance of the layered structure of the joint cartilage,surface cell structure damage and disordered arrangement,accompanied by a large amount of inflammatory cell infiltration,and an obvious loss of chondropolysaccharides,the expression levels of TNF-α,IL-6,IL-1β,and MDA in rat joint lavage fluid,the Mankin score,the the expression levels of NF-κB p65,NLRP3,and MMP-13 in knee joint tissue were higher(P＜0.05),the expression levels of SOD and GSH-PX in the lavage fluid and the expression level of COL-Ⅱ in the knee joint tissue were lower(P＜0.05).Compared with the model group,the degree of joint cartilage tissue damage in rats in the PHL-L group,PHL-M group,and PHL-H group gradually decreased,the joint cartilage layer gradually thickened,the four layer structure became clear,the number of bone cells increased and the arrangement gradually tended to be orderly,and the chondropolysaccharides gradually increased,the expression levels of TNF-α,IL-6,IL-1β,and MDA in rat joint lavage fluid,the Mankin score,the the expression levels of NF-κB p65,NLRP3,and MMP-13 in knee joint tissue gradually decreased(P＜0.05),the expression levels of SOD and GSH-PX in the lavage fluid and the expression level of COL-Ⅱ in the knee joint tissue gradually increased(P＜0.05).The addition of NF-κB pathway activator PMA reversed the improvement effect of PHL on osteoarthritis(P＜0.05).The results indicate that PHL can improve osteoarthritis induced by anterior cruciate ligament transection in rats,and its mechanism of action may be related to the inhibition of the NF-κB p65/NLRP3 signaling pathway.
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