膝三针调节PPARγ/NF-κB通路对膝骨关节炎大鼠骨代谢及骨微结构的影响OA
The effects of knee three needles on bone metabolism and microstructure in knee osteoarthritis rats by regulating PPARγ/NF-κB pathway
目的:探讨膝三针调节过氧化物酶体增殖物激活受体γ(PPARγ)/核因子κB(NF-κB)通路对膝骨关节炎(KOA)大鼠骨代谢及骨微结构的影响.方法:关节腔内注射碘乙酸钠溶液构建 KOA大鼠,采用随机数字表法将建模成功的大鼠随机分为 KOA 组、膝三针组(膝三针治疗)、膝三针+GW9662组(膝三针治疗+尾静脉注射1mg/kg GW9662),每组12只,另选择12只为 NC组,NC组和 KOA组尾静脉注射等量生理盐水,1次/d,每周连续5 d,共治疗3周.检测各组的疼痛行为学[机械痛阈值(PWT)、热痛阈值(PWL)];酶联免疫吸附测定(ELISA)检测血清中[肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6]和骨代谢指标[Ⅰ型胶原C末端肽(CTX-Ⅰ)、骨保护素(OPG)、抗酒石酸酸性磷酸酶(TRACP)、核因子κB受体活化因子配体(RANKL)]的 表 达;Micro-CT分析大鼠膝关节骨 微 结 构[骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)、骨小梁结构模式参数(SMI)、骨小梁间距(Tb.Sp)、骨密度(BMD)、骨体积分数(BV/TV)]的改变;苏木精伊红(HE)和甲苯胺蓝染色检测膝关节软骨组织病理变化;蛋白质印迹(Western Blot)检测膝关节软骨组织中PPARγ/NF-κB通路蛋白的表达.结果:NC组软骨组织各层细胞排列整齐、分布均匀;KOA组软骨组织表层不平整,可见细胞减少、排列紊乱、大量空泡;膝三针组软骨组织表层较为平整,细胞数量增多,空泡减少;膝三针+GW9662组软骨组织表层进一步不平整,细胞数量进一步减少,空泡进一步增 多.与 NC组相比,KOA 组 PWL、PWT、OPG、Tb.Th、Tb.N、BMD、BV/TV、PPARγ降低,Mankin评分、CTX-Ⅰ、TRACP、RANKL、TNF-α、IL-1β、IL-6、SMI、Tb.Sp、p-NF-κB P65/NF-κB P65升高(P<0.05);与 KOA 组相比,膝三针组 PWL、PWT、OPG、Tb.Th、Tb.N、BMD、BV/TV、PPARγ升 高,Mankin评分、CTX-Ⅰ、TRACP、RANKL、TNF-α、IL-1β、IL-6、SMI、Tb.Sp、p-NF-κB P65/NF-κB P65降低(P<0.05);膝三针+GW9662组可以逆转膝三针组上述因子的变化(P<0.05).结论:膝三针通过激活PPARγ进而抑制 NF-κB改善KOA大鼠的骨代谢及骨微结构.
Objective:To explore the effects of knee three needles on bone metabolism and microstructure in knee osteoarthritis(KOA)rats by regulating the peroxisome proliferator-activated receptor gamma(PPARγ)/nuclear factor kappa B(NF-κB)pathway.Methods:Sodium iodoacetate solution was injected into the joint cavity to con-struct KOA rats.The successfully modeled rats were assigned into KOA group,knee three needles group(knee three needles treatment),and knee three needles+GW9662 group(knee three needles treatment+tail vein injection of 1 mg/kg GW9662),with 12 rats per group.There were 12 animals in the normal control(NC)group.The NC group and KOA group received intravenous in-jection of an equal amount of physiological saline once a day for 5 consecutive days per week,for a total of 3 weeks of treatment.The pain behavior of each group was tested[paw withdrawal threshold(PWT),paw withdrawal latency(PWL)].The enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6,and bone metabolism indicators[type I collagen C-terminal peptide(CTX-Ⅰ),osteoprotegerin(OPG),tartrate-resistant acid phosphatase(TRACP),and receptor activator of nuclear factor κB ligand(RANKL)].Micro-CT was used to analyze the changes in the bone microstructure of the rat knee joint[including trabecular thickness(Tb.Th),trabecular number(Tb.N),structure model index(SMI),trabecular separation(Tb.Sp),bone mineral density(BMD),and bone volume/tissue volume ratio(BV/TV)].Hematoxylin and eosin(HE)staining and toluidine blue staining were implemented to measure pathological changes in knee cartilage tissue.In addition,Western blot was used to measure the expression of PPARγ/NF-κB pathway proteins in knee cartilage tissue.Results:The cells in each layer of cartilage tissue in the NC group were arranged neatly and distributed evenly.The surface of the cartilage tissue in the KOA group was un-even,with reduced cells,disordered arrangement,and a large number of vacuoles visible.The surface of the carti-lage tissue in the knee three needles group was relatively flat,with an increase in the number of cells and a decrease in vacuoles.The surface of the cartilage tissue in the knee three needles+GW9662 group was uneven,with a further decrease in cell number and an increase in vacuoles.Compared with the NC group,the KOA group showed decrea-ses in PWL,PWT,OPG,Tb.Th,Tb.N,BMD,BV/TV,and PPARγ,along with increases in Mankin score,CTX-Ⅰ,TRACP,RANKL,TNF-α,IL-1β,IL-6,SMI,Tb.Sp,and the ratio of p-NF-κB P65 to NF-κB P65(P<0.05).Compared with the KOA group,the knee three needles group exhibited increases in PWL,PWT,OPG,Tb.Th,Tb.N,BMD,BV/TV,and PPARγ,and decreases in Mankin score,CTX-Ⅰ,TRACP,RANKL,TNF-α,IL-1β,IL-6,SMI,Tb.Sp,and the p-NF-κB P65/NF-κB P65 ratio(P<0.05).The knee three needles+GW9662 group reversed the changes in the above factors induced by the knee three-needle acupuncture intervention(P<0.05).Conclusion:Knee acupuncture improves bone metabolism and microstructure in KOA rats by activating the PPARγ/NF-κB pathway.
衣美霞;张蝶;王宁
海军第九七一医院康复医学与理疗科,山东 青岛 266071海军第九七一医院中医科海军第九七一医院中医科
医药卫生
膝三针过氧化物酶体增殖物激活受体γ核因子κB膝骨关节炎骨代谢骨微结构
knee three needlesperoxisome proliferator-activated receptor gammanuclear factor kappa Bknee osteoarthritisbone metabolismbone microstructure
《中国康复》 2026 (6)
323-329,7
2023年度军队中医药服务能力培育与提升专项计划(2023ZY025)
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