中医整脊棘突旁指压法干预大鼠膝骨关节炎的效果及作用机制研究  

Efficacy and mechanism of traditional Chinese medicine chiropractic paraspinous thumb-pressing therapy against knee osteoarthritis in rats:an experimental study

作　　者：张力 张华[1] 李平[1] 高嵩[1] 柯广娟 屈留新[1] ZHANG Li;ZHANG Hua;LI Ping;GAO Song;KE Guangjuan;QU Liuxin(Zhongda Hospital Affiliated to Southeast University,Nanjing 210009,Jiangsu,China)

机构地区：[1]东南大学附属中大医院,江苏南京210009

出　　处：《中医正骨》2025年第3期14-22,38,共10页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：东南大学附属中大医院中西医协同医学科研项目(2023zxyxt03)。

摘　　要：目的:探讨中医整脊棘突旁指压法干预大鼠膝骨关节炎(knee osteoarthritis,KOA)的效果及作用机制。方法:将40只SD大鼠随机分为空白组、模型组、低强度组、中强度组和高强度组,每组8只。模型组、低强度组、中强度组和高强度组大鼠均采用碘乙酸钠关节腔注射建立双侧KOA模型,空白组大鼠双侧膝关节腔注射等量生理盐水。造模14 d后,采用小动物计力指压器于L_(3)~L_(5)棘突旁,对低、中、高强度组大鼠分别施以4 N、8 N、16 N的垂直向下压力,模拟不同力度的棘突旁指压法;每次按压2 s,每处连续按压2 min;每隔1 d施术1次,干预28 d;空白组和模型组不进行任何干预。分别于干预1 d、7 d、14 d、21 d、28 d,采用von Frey电子测痛仪测定大鼠机械刺激缩足阈值(paw withdrawal mechanical threshold,PWMT)。干预28 d后,采用ELISA试剂盒检测大鼠血清中神经生长因子(nerve growth factor,NGF)、降钙素基因相关肽(calcitonin gene related peptide,CGRP)含量;采用钙成像技术检测大鼠L_(3)背根神经节神经元Ca^(2+)通量;采用Western blot法检测大鼠L_(3)背根神经节组织中瞬时受体电位香草素受体1(transient receptor potential vanilloid 1,TRPV1)、脂蛋白磷酸酶2B(protein phosphatases 2B,PP2B)、钙调蛋白(calmodulin,CaM)的蛋白表达水平;分别采用HE染色试剂盒、Masson染色试剂盒、天狼星红染色试剂盒对大鼠膝关节滑膜组织进行染色,观察滑膜组织的炎性细胞浸润和纤维化情况;采用Western blot法检测大鼠膝关节滑膜组织中白细胞介素(interleukin,IL)-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、转化生长因子(transforming growth factor,TGF)-β、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的蛋白表达水平。结果:①PWMT测定结果。空白组、模型组、低强度组大鼠PWMT随时间变化均基本保持不变(F=0.885,P=0.487;F=0.339,P=0.849;F=0.555,P=0.697),中、高强度组大鼠PWMT随时间变�Objective:To observe the effects of traditional Chinese medicine(TCM)chiropractic paraspinous thumb-pressing therapy against knee osteoarthritis(KOA)in rats,and to explore its underlying mechanism.Methods:Forty SD rats were selected and randomized into blank group,model group,low-intensity group,medium-intensity group and high-intensity group,with 8 ones in each group.All rats but the ones in blank group were modeled by using bilateral knee intra-articular injection of sodium iodoacetate for inducing KOA,while the ones in blank group by using injection of the same dosage of normal saline at the corresponding sites.Fourteen days after successful modeling,the rats in the low-,medium-,and high-intensity groups were subjected to 4 N,8 N,and 16 N vertical downward pressures,respectively,beside the L_(3)-L_(5) spinous processes by using a force-measuring finger pressure device for small animals to simulate the paraspinous thumb-pressing manipulation with different intensities,once every other day,2 seconds per time and 2 minutes per site for consecutive 28 days;while the ones in blank group and model group were not given any intervention.The paw withdrawal mechanical threshold(PWMT)of rats were tested using a Von Frey electronic algometer after 1-,7-,14-,21-and 28-day intervention,respectively.After 28-day intervention,the levels of nerve growth factor(NGF)and calcitonin gene related peptide(CGRP)in serum were detected by using ELISA kit,and the Ca^(2+)flux in L_(3) dorsal root ganglion(DRG)neurons was detected by using Calcium imaging.Furthermore,the protein expression levels of transient receptor potential vanilloid 1(TRPV1),protein phosphatases 2B(PP2B)and calmodulin(CaM)in the L_(3) DRG tissues were detected using Western blot.After that,the rat knee synovial tissues were harvested and stained with HE staining kit,Masson staining kit,and Sirius red staining kit,respectively,to observe the inflammatory cell infiltration and fibrosis,and the protein expression levels of interleukin(IL)-1β,tumor necrosis factor-α(TN

关 键 词：骨关节炎 膝 中医整脊 指压法 疼痛 敏化 瞬时受体电位香草素受体1 神经节 脊 神经元 滑膜 成纤维细胞 串话 大鼠 

分 类 号：R274[医药卫生—中医骨伤科学]