低频脉冲电磁场联合中药对膝骨关节炎小鼠MMP-13和Col-X的影响  

Effects of low-frequency pulsed electromagnetic field combined with traditional Chinese medicine on MMP-13 and Col-X in mice with knee osteoarthritis

作　　者：李心芸 熊振飞[2] 吴杏英 汤样华[2] LI Xinyun;XIONG Zhenfei;WU Xingying;TANG Yanghua(Department of Rehabilitation,First People’s Hospital of Xiaoshan District,Hangzhou 311200,China;Department of Orthopedic,Xiaoshan District Hospital of Traditional Chinese Medicine,Hangzhou 311201,China;The Third Clinical Medicine College of Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]杭州市萧山区第一人民医院康复科,浙江杭州311200 [2]杭州市萧山区中医院骨科,浙江杭州311201 [3]浙江中医药大学第三临床医学院,浙江杭州310053

出　　处：《健康研究》2023年第6期666-671,F0003,共7页Health Research

基　　金：萧山区社会发展重大科技计划政策引导项目(2018304);杭州市农业与社会发展科研引导项目(20220919Y082)。

摘　　要：目的从分子生物学、形态学角度,对比观察低频脉冲电磁场联合中药和单纯中药治疗膝关节骨性关节炎(knee osteoarthritis,KOA)小鼠的效果。方法采用Hulth法建立KOA模型小鼠40只,造模成功后分为空白对照组、中药组、低频脉冲电磁场组、联合组,每组各10只。空白对照组和中药组分别予生理盐水、益气补肾化瘀方水煎剂灌胃,每次1 mL,每日2次;低频脉冲电磁场组予低频脉冲电磁场治疗,每天1次,每次30 min;联合组给予中药灌胃的基础上加用低频脉冲电磁场。连续处理2个月后处死小鼠,取膝关节,采用ELISA、qR-TPCR、免疫组化、番红-固绿染色及Micro-CT观察小鼠关节软骨组织功能和形态变化。结果ELISA检测显示,造模后的小鼠血清中IL-1β和TNF-α含量均显著升高,模型制备成功。qRT-PCR和免疫组化结果显示,中药组、低频脉冲电磁场组、联合组的膝关节组织中基质金属蛋白酶-13(MMP-13)、X型胶原蛋白(Col-X)表达水平,及膝关节软骨组织中MMP-13表达量均低于空白组,且联合组最低,差异均有统计学意义(P<0.05);Micro-CT结果显示,联合组小鼠膝关节骨体积分数与骨小梁厚度均高于中药组,差异有统计学意义(P<0.01),2组间骨小梁分离度与骨体积差异无统计学意义(P>0.05)。结论低频脉冲电磁场联合益气补肾化瘀方可降低KOA小鼠膝关节组织中Col-X与MMP-13的表达水平,改善小鼠关节软骨组织功能和形态。Objective To conduct a comparative investigation on the molecular biology and morphology perspectives,examining the effects of low-frequency pulsed electromagnetic field combined with traditional Chinese medicine(TCM)versus TCM alone in the treatment of knee osteoarthritis(KOA)mice.Methods The Hulth method was used to create 40 models of mice with KOA.These mice were then divided into four groups-the blank control group,traditional Chinese medicine group,low-frequency pulsed electromagnetic field group,and combined group-each consisting of 10 mice,after a successful modelling.Mice in the control group received 1ml of normal saline via gavage twice daily,whilst the Yiqi Bushen Huayu Therapy group received the same treatment.The low frequency pulsed electromagnetic fields group was exposed to this therapy once daily for 30 minutes on their knee joint.The combined group received herbal gavage with the incorporation of low-frequency pulsed electromagnetic fields.Following two months of continuous treatment,the mice were euthanized and their knee joints were extracted for analyzing the functional and morphological variations in their articular cartilage tissue via Micro-CT,ELISA,qR-TPCR,immunohistochemistry,and fanconiosis-solidus green staining.Results The ELISA assay demonstrated a significant increase in serum levels of IL-1β and TNF-α in mice following modelling,indicating the successful preparation of the model.qRT-PCR and immunohistochemistry results revealed lower expression levels of matrix metalloproteinase-13(MMP-13)and collagen type X(Col-X)in knee tissues of the traditional Chinese medicine group,the low-frequency pulsed electromagnetic field group,and the combined group as well as lower expression levels of MMP-13 in knee cartilage tissue compared to the blank group.Notably,the combined group exhibited the lowest expression level.Expression levels of MMP13 in knee joint tissues were lower compared to the blank group,with the lowest levels seen in the combined group.These differences were statistically s

关 键 词：膝关节骨性关节炎 基质金属蛋白酶-13 X型胶原蛋白 低频脉冲电磁场 益气补肾化瘀方 

分 类 号：R274[医药卫生—中医骨伤科学]