摘要
背景:在前期应用中发现疏调经筋推拿法能有效改善膝骨关节炎的临床症状,但其治疗机制有待验证。目的:探讨疏调经筋推拿法对膝骨关节炎的干预作用及其可能的作用机制。方法:34只雄性新西兰兔随机分为空白组(9只)及造模组(25只),造模组膝关节腔内注射4%木瓜蛋白酶溶液建立膝骨关节炎兔模型。6周后两组各随机挑选1只进行软骨苏木精-伊红染色及Mankin’s评分,模型评估后,造模组随机分为模型对照组、推拿组及氨糖组,每组8只。推拿组采取“疏调经筋推拿法”,隔日1次;氨糖组采用硫酸氨基葡萄糖胶囊水溶液灌胃,每日1次;共干预4周。干预结束1周后,番红O-固绿染色镜下观察软骨病理形态;Western blot、免疫组织化学染色法检测软骨组织中细胞外信号调节激酶1/2、核转录因子κB p65、Bcl-2、Bax、半胱氨酸天冬氨酸蛋白酶3蛋白表达水平;ELISA法测定外周血清和关节液中白细胞介素1β的质量浓度。结果与结论:①软骨切片镜下观察:模型对照组软骨表面缺损、破坏,软骨层变薄,软骨细胞异常簇状聚集,潮线破坏;推拿组软骨表面无明显缺损、破坏,软骨细胞排列基本正常,无明显异常聚集,潮线较规整;氨糖组与模型对照组相比未见明显改善;②与空白组相比,模型对照组细胞外信号调节激酶1/2、核转录因子κB p65、Bax、半胱氨酸天冬氨酸蛋白酶3蛋白表达水平升高(P<0.01),Bcl-2表达降低(P<0.01);推拿组兔细胞外信号调节激酶1/2、核转录因子κB p65、Bax、半胱氨酸天冬氨酸蛋白酶3蛋白表达水平明显低于模型对照组(P<0.01),稍低于氨糖组(P<0.05);Bcl-2表达明显高于模型对照组(P<0.01),稍高于氨糖组(P<0.05);③与空白组相比,模型对照组关节液、外周血清中白细胞介素1β质量浓度显著增高(P<0.01);与模型对照组相比,推拿组白细胞介素1β质量浓度降低(P<0.01);氨糖组白细胞介素1β质量浓度高于推拿组(P<0.01);④提示“疏调经筋推拿法”可以抑制软骨退变,延缓膝骨关节炎进展,可能是通过调节白细胞介素1β的分泌及释放、并通过白细胞介素1β与细胞外信号调节激酶1/2-核转录因子κB信号通路相互作用、抑制软骨细胞过度凋亡、维持软骨细胞内环境稳定、修复损伤软骨来实现的。
BACKGROUND:It has been found that Shutiao Jingjin Massage can effectively improve the clinical symptoms of knee osteoarthritis in the early stage,but its treatment mechanism needs to be verified.OBJECTIVE:To explore the effect of Shutiao Jingjin Massage on knee osteoarthritis and its possible mechanism METHODS:Thirty-four male New Zealand rabbits were randomly divided into blank group(n=9)and model group(n=25).Rabbit models of knee osteoarthritis were established by injecting 4%papain solution into the knee joint cavity.Six weeks after modeling,one rabbit in each group was randomly selected for cartilage hematoxylin-eosin staining and Mankin’s scoring.After the model evaluation,the model group was randomly divided into model control group(n=8),massage group(n=8)and glucosamine group(n=8).The massage group was treated with Shutiao Jingjin Massage,once every other day;the glucosamine group was given aqueous solution of glucosamine sulfate capsules by gavage,once a day.Interventions in each group lasted 4 weeks.At 1 week after intervention,the pathological morphology of cartilage was observed by safranin O-fast green staining.The expression of extracellular signal-regulated kinases 1 and 2,nuclear factor-kappa B p65,Bcl-2,Bax,and cysteine-aspartic acid protease-3 in cartilage tissue was detected by western blot assay and immunohistochemistry.The levels of interleukin-1βin peripheral blood and joint fluid were determined by enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:Cartilage section microscopic observation:the model control group had cartilage surface defect and damage,cartilage layer thinning,abnormal cluster aggregation of chondrocytes,and tidal line damage;in the massage group,there was no obvious damage on the cartilage surface,chondrocyte arrangement was basically normal,no obvious abnormal aggregation occurred,and the tidal line was regular;compared with the model control group,the glucosamine group did not show significant improvement.Compared with the blank group,the expression of extracellular signal-regulated kinases 1 and 2,nuclear factor-kappa B p65,Bax,and cysteine-aspartic acid protease-3 protein in the model control group was increased(P<0.01),while the Bcl-2 expression decreased(P<0.01).The expression levels of extracellular signal-regulated kinases 1 and 2,nuclear factor-kappa B p65,Bax,and cysteineaspartic acid protease-3 protein in the massage group were significantly lower than those in the model control group(P<0.01)and slightly lower than those in the glucosamine group(P<0.05),while the expression of Bcl-2 was significantly higher than that in the model control group(P<0.01)and slightly higher than that in the glucosamine group(P<0.05).Compared with the blank group,the mass concentrations of interleukin-1βin the joint fluid and peripheral blood were significantly increased in the model control group(P<0.01).Compared with the model control group,the mass concentrations of interleukin-1βin the joint fluid and peripheral blood were significantly decreased in the massage group(P<0.01).The mass concentrations of interleukin-1βin the joint fluid and peripheral blood were significantly higher in the glucosamine group than the massage group(P<0.01).To conclude,Shutiao Jingjin Massage can inhibit cartilage degeneration and slow down the progression of knee osteoarthritis,probably by regulating the secretion and release of interleukin-1β,interacting with the extracellular signal-regulated kinase 1/2-nuclear transcription factorκB signaling pathway,inhibiting excessive apoptosis of chondrocytes,maintaining the stability of intracellular environment in chondrocytes and repairing damaged cartilage.
作者
郑利君
王凯
李牧真
王建民
乔英杰
李华东
Zheng Lijun;Wang Kai;Li Muzhen;Wang Jianmin;Qiao Yingjie;Li Huadong(Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong Province,China;Department of Massage,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第35期5681-5687,共7页
Chinese Journal of Tissue Engineering Research
基金
山东省中医药科技项目(2021Q080),项目负责人:李牧真
齐鲁医派中医学术流派传承项目[鲁卫函(2022)93号],项目负责人:李华东
关键词
膝骨关节炎
疏调经筋
软骨损伤
细胞外信号调节激酶1/2
核转录因子ΚB
软骨内环境
knee osteoarthritis
Shutiao Jingjin
cartilage injury
extracellular signal-regulated kinase 1/2
nuclear transcription factorκB
cartilage internal environment
