调和阴阳针刀对膝骨关节炎大鼠关节功能及下肢力学的改善作用OA北大核心
Effects of harmonizing needle-knife therapy on joint function and lower limb mechanics in rats with knee osteoarthritis
目的 观察调和阴阳针刀干预对膝骨关节炎(knee osteoarthritis,KOA)模型大鼠膝关节功能及下肢力学的影响,探讨针刀治疗KOA的效应机制.方法 将 40 只SD大鼠随机分为空白组、模型组、针刀组和药物组.除空白组外均于左侧膝关节腔注射 4%木瓜蛋白酶与 0.03 mol/L L-半胱氨酸混合溶液建立KOA模型.针刀组大鼠采用调和阴阳针刀法,在内、外侧副韧带及髌韧带进行操作,药物组大鼠每日通过灌胃方式连续给予塞来昔布(10 mg/(kg·d)).针刀组与药物组大鼠分别在造模后第 7 天进行干预,每周 1 次,持续 4周.所有大鼠干预前及干预结束 1 周后参照改良Lequesne MG膝关节级别评估法评分并完成转棒疲劳实验.干预结束 1 周后进行活体DR检查膝关节腔隙,经苏木素-伊红(HE)、番红固绿染色观察软骨细胞形态,行韧带力学拉伸测试,分别取血清、软骨组织采用 ELISA、Western Blot 法检测基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)蛋白表达.结果 (1)与空白组相比,模型组大鼠改良Lequesne MG的膝关节级别评分增高,转棒疲劳实验时长下降,关节腔隙减小(P<0.01);(2)与模型组、药物组相比,针刀组大鼠改良Lequesne MG的膝关节级别评分降低,转棒疲劳实验时长上升(P<0.05),膝关节的关节腔间隙增大(P<0.05);(3)针刀组大鼠韧带弹性模量与模型组相比,无显著性差异,与药物组相比增高.与模型组、药物组相比,针刀组大鼠屈服强度、最大应变、屈强比增高(P<0.05);(4)HE和番红固绿染色显示,与模型组相比,针刀组仅有少量炎细胞浸润;与药物组相比,针刀组大鼠软骨表面较为平整,仅有轻微不连续;(5)ELISA检测,与模型组相比,针刀组大鼠血清MMP-13 含量下降(P<0.01);药物组和针刀组大鼠血清MMP-13 无明显差异;(6)与模型组相比,针刀组大鼠软骨MMP-13 含量显著降低(P<0.01);与药物组相比,针刀组大鼠含量下降(P<0.05).结论 针刀干预可以提升KOA模型大鼠膝关节稳定性,改善下肢力学结构,减少MMP-13表达.
Objective This study aimed to observe the effects of Yin-Yang balancing acupotomy intervention on knee-joint function and lower limb biomechanics in a rat model of knee osteoarthritis(KOA),and to explore the mechanisms of acupotomy when treating KOA.Methods Forty SD rats were randomly divided into a blank group,a model group,an acupotomy group,and a medication group.Except for the blank group,KOA models were established by injecting a mixed solution of 4%papain and 0.03 mol/L L-cysteine into the left knee-joint cavity.The acupotomy group received Yin-Yang balancing acupotomy interventions targeting the medial/lateral collateral ligaments and patellar ligament.The medication group received daily oral celecoxib(10 mg/(kg·d)).Interventions began on day 7 post-modeling,and occurred once weekly for 4 weeks.All rats were assessed pre-and post-intervention using the modified Lequesne MG knee-joint grading system and rotarod fatigue test.Post-intervention,in vivo DR imaging was used to measure joint space width.Cartilage morphology was evaluated via HE and safranin O-fast green staining.Ligament biomechanical tensile testing was performed.Serum and cartilage tissues were analyzed by ELISA and Western Blot for matrix metalloproteinase-13(MMP-13)expression.Results(1)Compared with the blank group,the model group showed increased modified Lequesne MG scores,reduced rotarod endurance time,and narrowed joint space(P<0.01).(2)Compared with the model and medication groups,the acupotomy group exhibited lower Lequesne MG scores,prolonged rotarod endurance time(P<0.05),and expanded joint space(P<0.05).(3)The elastic modulus of ligaments in the acupotomy group showed no significant difference from those in the model group but was higher than those in the medication group.Yield strength,maximum strain,and yield-to-tensile strength ratio in the acupotomy group were higher than those in the model and medication groups(P<0.05).(4)HE and Safranin O-Fast green staining revealed minimal inflammatory infiltration in the acupotomy group compared with the model group.Cartilage surfaces in the acupotomy group were smoother than those in the medication group.(5)ELISA showed reduced serum MMP-13 levels in the acupotomy group versus the model group(P<0.01),and no significant differences between levels in the drug and acupotomy groups.(6)Cartilage MMP-13 expression in the acupotomy group was significantly lower than that in the model group(P<0.01)and lower than that in the medication group(P<0.05).Conclusions Acupotomy intervention enhances knee joint stability,improves lower limb mechanical alignment,and suppresses MMP-13 expression in KOA rats.
徐懿;冼乐瑶;林煜翔;许铛瀚;罗孟西;郑谅
广州中医药大学第一临床医学院,广州 510405广州中医药大学第一临床医学院,广州 510405广州中医药大学第一临床医学院,广州 510405广州中医药大学第一临床医学院,广州 510405广州中医药大学第一附属医院针推康复中心,广州 510405广州中医药大学第一附属医院针推康复中心,广州 510405
生物科学
膝骨关节炎针刀MMP-13下肢力学软骨细胞
knee osteoarthritisneedle-knifeMMP-13lower limb mechanicschondrocytes
《中国实验动物学报》 2025 (7)
1010-1020,11
广东省中医药局科研项目(20231124),国家中医药局区域中医诊疗中心(针灸)建设项目.Funded by Research Project of Guangdong Provincial Administration of Traditional Chinese Medicine(20231124),Construction Project of Regional TCM Medical Center(Specializing in Acupuncture)Under the National Administration of Traditional Chinese Medicine.
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