火针对膝骨关节炎大鼠关节软骨MMP-3、TGF-β1、TNF-α的影响

Effect of heated needle on articular cartilage MMP-3,TGF-β1 and TNF-α in rats with knee osteoarthritis

目的:探讨火针对膝骨关节炎大鼠关节软骨MMP-3、TGF-β1、TNF-α的影响。方法:20只健康SD雄性大鼠,剃去后肢髋关节至脚趾毛,踝关节背屈70～90度,伸展膝关节至160～180度,纱布固定,将石膏从腹股沟至脚趾缠绕大鼠后肢,脚趾露出,绷带固定,建立KOA大鼠模型,另外10只不作任何处理作为正常组。六周后模型制作成功。20只模型大鼠随机分为2组,模型组和火针组。火针组选取膝前穴、阿是穴和阳陵泉穴,进行火针治疗,每3 d行一次针,共治疗6次。治疗前后对各组大鼠进行Lequesne MG评分评估,全自动生物化学分析仪测定各组大鼠血清MMP-3、TGF-β1、TNF-α的表达水平,HE染色后观察大鼠膝关节软骨病理形态变化,采用放射免疫分析法检测大鼠软骨组织中MMP-3、TGF-β1、TNF-α含量。结果:(1)与对照组比较,模型组大鼠血清MMP-3、TGF-β1和TNF-α表达水平显著增高,差异有统计学意义(P<0. 05)。(2)模型组大鼠膝骨关节软骨表面粗糙,软骨细胞肥大,表层细胞坏死,巢状增生,潮线消失,呈明显退行性病变。(3)与对照组比较,模型组大鼠膝关节软骨组织中MMP-3、TGF-β1和TNF-α含量均显著升高(P<0. 05)。(4)与模型组比较,火针组大鼠血清MMP-3、TGF-β1和TNF-α表达水平均明显降低,差异有统计学意义(P<0. 05)。(5)火针组大鼠膝骨关节软骨表面较光滑,细胞排列规则,表层细胞轻度增生,潮线较完整。(6)与模型组比较,火针组大鼠膝关节软骨组织MMP-3、TGF-β1和TNF-α含量均明显降低,差异有统计学意义(P<0. 05)。结论:火针治疗可有效减轻大鼠膝关节软骨损伤,降低血清及膝关节软骨组织中MMP-3、TGF-β1和TNF-α水平,进而发挥抗炎作用,可能是火针治疗膝骨关节炎的一个作用机制。

Objective:To investigate the effects of heated needle on the articular cartilage MMP-3,TGF-β1 and TNF-αin rats with knee osteoarthritis.Methods:Twenty healthy male Sprague-Dawley rats were shaved to remove the hips from the hip to the toe of the hind legs.The ankles were dorsiflexed at 70-90 degrees and the knee was stretched to 160-180 degrees.The gauze was fixed and the gypsum was wound around the hind limbs from the inguinal to the toe,exposed toes,fixed bandages,established KOA rat model,the other 10 were not treated as normal group.Six weeks later the model was made.Twenty rats were randomly divided into two groups:model group and fire needle group.Heated needle group selected knee ankle,acupoints and Yanglingquan,fire needle treatment,once every 3 d line needle,a total of 6 times treatment.The Lequesne MG scores were evaluated before and after treatment.The expression levels of serum MMP-3,TGF-β1 and TNF-αwere measured by automatic biochemical analyzer.The pathology of knee joint cartilage was observed after HE staining.Morphological changes were used to detect the contents of MMP-3,TGF-β1 and TNF-αin rat cartilage tissue by radioimmunoassay.Results:①Compared with the control group,the expression levels of MMP-3,TGF-β1 and TNF-αin the model group were significantly increased,the difference was statistically significant(P<0.05).②In the model group,the articular cartilage of the knee joint showed rough surface,chondrocyte hypertrophy,necrosis of superficial cells,nested hyperplasia and disappearance of tide lines,showing obvious degenerative lesions.③Compared with the control group,the content of MMP-3,TGF-β1 and TNF-α in the model group rats′knee joint cartilage were significantly increased(P<0.05).④Compared with the model group,the expression levels of serum MMP-3,TGF-β1 and TNF-α in heated needle were significantly decreased,the difference was statistically significant(P<0.05).⑤The surface of knee joint cartilage of heated needle was smoother,the arrangement of cells was regular,the surface cells were slightly hyperplastic and the tide line was more complete.⑥Compared with the model group,the content of MMP-3,TGF-β1 and TNF-α in the articular cartilage of the rats in heated needle group were significantly decreased,the difference was statistically significant(P<0.05).Conclusion:Fire-acupuncture can effectively reduce the level of MMP-3,TGF-β1 and TNF-α in serum and knee cartilage of knee joint.It is possible that fire-acupuncture treatment may be a mechanism of fire-acupuncture treatment of knee osteoarthritis.