活血利水方干预膝骨关节炎大鼠的疗效及对NF-κB/HIF-1α/VEGF信号通路的调控作用OA

Objective:This study aims to investigate the thera-peutic effects and underlying mechanisms of the Huoxue Lishui formula in treating knee osteoarthritis(KOA)through regulation of the NF-κB/HIF-1α/VEGF signaling pathway.Methods:Thirty-six SD rats were divided into Chinese medi-cine group,Western medicine group,model group,and sham-operated group,with 9 rats in each group.KOA models were established in the right hind limbs of rats in the model group,the Chinese medicine group,and the Western medicine group using the Hulth surgery procedure.For the sham-operated group,only the medial skin and subcutaneous tissue of the right knee joint were incised to expose the joint cavity,which was then sutured.After successful modeling,rats in the Chinese medicine group were administered the Huoxue Lishui formula via oral gavage at a crude drug dose of 6 g/(kg·d),the Western medicine group received celecoxib solution at 12 mg/(kg·d)via oral gavage,and the model group and sham-operated group were given normal saline at 10 mL/(kg·d)via oral gavage.The intervention lasted for 8 weeks.Blood,knee joint cartilage,and synovial tissues were collected from each group.Pathological changes were observed using HE staining,serum levels of TNF-α,HIF-1α,and VEGF were detected by ELISA,and the protein and mRNA expression lev-els of NF-κB/HIF-1α/VEGF pathway-related factors were measured by Western Blot and qRT-PCR,respectively.Results:Compared with the sham-operated group,the model group exhibited significantly increased serum levels of TNF-α,HIF-1α,and VEGF(all P<0.001).Huoxue Lishui formula treatment significantly reduced the levels of all three factors(TNF-α,P=0.001;HIF-1α,P<0.001;VEGF,P<0.001),while the Western medicine group showed a significant reduction only in VEGF(P<0.001),with no significant effects on TNF-α(P=0.435)or HIF-1α(P=0.905).The VEGF content in the Chinese medicine group was lower than that in the Western medicine group,with statistical signifi-cance(P<0.001).At the molecular level,mRNA expression of NF-κB,HIF-1α,and VEGF was significantly upregulated in the model group(all P<0.01).Huoxue Lishui formula downregulated the mRNA expression of all three genes(all P<0.001),outperforming the Western medicine group in suppressing NF-κB(P=0.023)and HIF-1α(P=0.045)mRNA expression.The Western medicine group had no significant effect on VEGF mRNA(P=0.511).In the model group,the protein levels of synovial NF-κB,HIF-1α,and VEGF were higher than those in the sham-operated group(P<0.001,P=0.029,P<0.001).Treatment with Chinese medicine significantly reduced all three proteins,whereas Western medicine reduced NF-κB/VEGF(P<0.05).Chinese medicine resulted in lower levels of NF-κB/VEGF compared with Western medicine(P<0.001).Histopathological results confirmed that Huoxue Lishui formula provided the most significant improvement in cartilage structure and exhibited the strongest anti-inflammatory effect.Conclusion:Huoxue Lishui formula may delay the progression of KOA by suppressing the activation of the NF-κB/HIF-1α/VEGF signaling pathway,thereby reducing synovial inflammation,knee joint inflammatory responses,and pathological neovascularization.