火针对膝骨关节炎大鼠关节功能及炎性反应的影响

Effect of fire-needle intervention on joint function,cartilage impairment and inflammatory response in knee osteoarthritis rats

目的:观察火针对膝骨关节炎(KOA)大鼠退化软骨和炎性因子的影响,探讨火针干预KOA的分子生物学机制。方法:将SD大鼠随机分为正常组、模型组、火针干预组,每组13只。采用右侧膝关节腔内注射单碘乙酸盐制备KOA大鼠模型。火针干预组选择右侧"内膝眼""犊鼻"穴火针点刺,每周2次,治疗3周。观察各组大鼠爪压和步态评分;采用番红固绿染色观察右膝软骨组织病理形态并行骨关节炎(OA)评分;透射电镜观察右膝软骨结构;采用酶联免疫吸附法检测血清白细胞介素1α(IL-1α)、IL-10、IL-37、肿瘤坏死因子-α(TNF-α)、转化生长因子β(TGF-β)的含量。结果:与正常组比较,模型组大鼠爪压和步态评分、OA评分、血清IL-1α和TNF-α含量均升高(P<0.05,P<0.01),血清IL-10、IL-37、TGF-β含量均降低(P<0.01)。火针干预组爪压和步态评分、OA评分、血清IL-1α和TNF-α含量较模型组均降低(P<0.05,P<0.01),血清IL-10、IL-37、TGF-β含量较模型组均升高(P<0.01)。模型组可见脂滴较正常组多且大,线粒体肿胀,甚至空泡,粗面内质网扩张,甚至断裂溶解,数量较正常组减少,排列杂乱;火针干预组可见到丰富的轻度扩张、排列相对较规则的粗面内质网和线粒体,脂滴增多。结论:火针可改善KOA大鼠运动功能和膝骨关节软骨形态,其机制可能与降低血清炎性因子、提高抗炎因子的表达水平有关。

Objective To observe the effect of fire-needle stimulation of"Neixiyan"(EX-LE4)and"Dubi"(ST35)on changes of motor function,structure of cartilage degradation and inflammatory factors in knee osteoarthritis(KOA)rats,so as to explore its underlying mechanisms in improving KOA.Methods Thirty-nine male SD rats were randomly divided into normal,model and fire-needle groups(n=13 in each group).The KOA model was established by injection of Monoiodoacetate(MIA,1 mg)into the lumen of the right knee joint.On the 7th day after successful modeling,fire-needle was applied to EX-LE4 and ST35,twice a week for 3 weeks.The rats’behavioral reactions of gait(0 to 3 points)and claw pressure(0 to 3 points)were scored,and histopathological changes scored by assessing the impairment grade(0 to 6)and stage(0 to 4)of the articular cartilage after safranin O-fast green staining.The contents of serum Interleukin(IL)-1α,Tumor necrosis factorα(TNF-α),IL-10,IL-37 and transforming growth factorβ(TGF-β)were assayed by ELISA,and ultrastructural changes were observed under transmission electron microscope(TEM).Results After modeling,the gait and claw-pressure scores were significantly increased in the model group in comparison with the normal group(P<0.05),and the OA score and contents of serum IL-1αand TNF-αwere also appa-rently increased in the model group relevant to the normal group(P<0.05,P<0.01),while the levels of serum IL-10,IL-37 and TGF-βremarkably decreased in the model group in contrast to the normal group(P<0.01).After the intervention,the increase of gait and claw-pressure and OA scores,as well as serum IL-1αand TNF-αcontents,and the decrease of serum IL-10,IL-37 and TGF-βlevels were all reversed in the fire needle group(P<0.05,P<0.01).Outcomes of TEM showed more and larger lipid droplets,swollen mitochondria with some vacuoles,and expanded,broken or dissolved rough endoplasmic reticulum in the model group,which was milder in the fire-needle group.Conclusion Fire-needle can improve motor function and relieve impairment of articular cartilage of KOA rats,which may be related to its effects in reducing inflammatory factors and in increasing anti-inflammatory factor levels.