理筋正骨手法联合热补针对新西兰大白兔膝关节炎软骨损伤、免疫平衡及肝X受体NF-κB通路的影响
Effects of Lijin Zhenggu Therapy combined with heat supplementing needles on cartilage damage, immune balance, and liver X receptor nuclear factor NF-κB pathway in knee arthritis of New Zealand white rabbits
摘要目的:探讨理筋正骨手法联合热补针对新西兰大白兔膝关节炎软骨损伤、免疫平衡及肝X受体α(LXRα)/NF-κB通路的影响。方法:将60只大白兔按随机数字表法分为正常组、模型组、理筋正骨组、热补针组、联合组,每组12只。除正常组外,其余各组建立膝关节炎模型,理筋正骨组采用理筋正骨手法干预,热补针组采用热补针干预,联合组采用理筋正骨手法联合热补针干预。采用测痛仪检测各组大白兔痛阈值;HE染色观察软骨损伤形态;ELISA法检测关节面软骨组织中前列腺素E 2(PGE 2)、IL-1β、β-内啡肽(β-EP)水平;PCR检测滑膜组织LXRα、NF-κB mRNA水平;Western blot法检测滑膜组织TLR4、髓样分化因子(MyD88)、干扰素调节因子-7(IRF-7)、NF-κB P65蛋白表达。 结果:与模型组比较,热补针组、理筋正骨组、联合组大白兔痛阈值升高( P<0.05);关节面软骨组织PGE 2、IL-1β水平降低( P<0.05),β-EP水平升高( P<0.05);滑膜组织LXRα mRNA水平升高( P<0.05),NF-κB mRNA水平降低( P<0.05);滑膜组织TLR4、MyD88、IRF-7、NF-κB P65蛋白表达降低( P<0.05)。 结论:理筋正骨手法联合热补针可降低膝关节炎模型大白兔PGE 2、IL-1β水平,提高β-EP水平,改善疼痛及软骨组织形态,其机制可能与调控LXRα/NF-κB通路,减少炎症反应,维持免疫平衡有关。
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abstractsObjective:To explore Lijin Zhenggu Therapy combined with heat supplementing needles on cartilage damage, immune balance, and liver X receptor NF-κB in knee osteoarthritis of New Zealand white rabbits.Methods:Totally 60 New Zealand white rabbits were divided into normal group, model group, Lijin Zhenggu Therapy group, heat supplementing needles group, and a combination group using random number table method, with 12 rabbits in each group. Except for the normal group, all other groups were established with knee osteoarthritis models. The Lijin Zhenggu Therapy group was treated with Lijin Zhenggu Therapy, the heat supplementing needles group was treated with heat supplementing needles, the combination group was treated with Lijin Zhenggu Therapy combined with heat supplementing needles. The pain threshold of each group of white rabbits was observed using a pain meter; HE staining was used to observe the morphology of cartilage damage in each group of white rabbits; ELISA was used to detect PGE 2, IL-1β and β-EP in the articular cartilage tissue of white rabbits in each group; PCR was used to detect the levels of LXRα and NF-κB mRNA. Western blot was used to detect toll like receptor 4 (TLR4), myeloid differentiation factor (MyD88), interferon regulatory factor-7 (IRF-7) in synovial tissue of rabbits in each group. Results:Compared with model group, the pain threshold of rabbits in heat supplementing needles group, Lijin Zhenggu Therapy group and combination group increased ( P<0.05); the levels of PGE 2 and IL-1β decreased ( P<0.05), while the level of β-EP increased ( P<0.05). LXRα mRNA level increased ( P<0.05), and NF-κB mRNA level decreased ( P<0.05); the expressions of TLR4, MyD88, IRF-7 and NF-κB P65 in synovial tissue decreased ( P<0.05). Conclusions:Lijin Zhenggu Therapy combined with heat supplementing needles can reduce the levels of PGE 2 and IL-1β, increase β-EP level, improve pain and cartilage tissue morphology. The mechanism may be related to the regulation of liver X receptor nuclear factor NF-κB pathway, reduction of inflammatory reactions and immunity maintaining.
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