Objective: To compare the effects of filiform needling, electroacupuncture (EA) and moxibustion on articular cartilage morphology and nuclear transcription factor-κB (NF-κB) p65/ Nod-like receptor protein 3 (NLRP3) pathway in rats with knee osteoarthritis (KOA).
Methods: Wistar rats were randomly divided into blank control， model, filiform needling, EA and moxibustion groups, with 8 rats in each group. The KOA model was established by injecting sodium iodoacetate into the right knee cavity. Filiform needling, EA and moxibustion groups were treated with the right side of "Dubi" (ST35) and "Neixiyan" (EX-LE5), and were given filiform needling, EA and mild moxibustion therapies for 15 min respectively, once every other day, for a total of 4 weeks. The diameter of the right knee joint was observed. The ultrastructure of knee chondrocytes was observed by transmission electron microscope. The contents of serum tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-18 were determined by ELISA. The expression levels of NF-κB p65, NLRP3, apoptosis-related speckle like protein (ASC) and TNF-α in knee cartilage were dectected by Western blot.
Results: Compared with the blank control group, the right knee joint cavity was narrowed, chondrocytes were constricted significantly, mitochondria were moderately swollen, the diameter of the right knee was increased, and the contents of serum TNF-α, IL-1β and IL-18 were increased (P<0.01), the expressions of NF-κB p65, NLRP3 and TNF-α in knee cartilage were increased (P<0.01) while the expression of ASC was decreased (P<0.01) in the model group.Compared with model group, the narrowed knee joint cavity and chondrocyte injury were improved， knee diameter decreased (P<0.01, P<0.05), and the content of serum TNF-α and the expression of NLRP3 in knee cartilage were decreased (P<0.05, P<0.01) in 3 treatment groups; the contents of serum IL-1β and IL-18, and the expression of TNF-α in knee cartilage were decreased (P<0.01, P<0.05), while the expression of ASC in knee cartilage was increased (P<0.01) in EA and moxibustion groups; the expression of NF-κB p65 in knee cartilage was decreased (P<0.01) in moxibustion group. Compared with EA group, the content of serum IL-18 was decreased (P<0.05) in moxibustion group.
Conclusion: Filiform needling, EA and moxibustion can all reduce the formation of knee osteophytosis in KOA rats, alleviate joint cavity narrowing, improve the ultrastructure of articular cartilage, reduce the level of inflammatory factors, regulate the NF-κB p65/NLRP3 pathway. Moxibustion has the most obvious regulatory effect among the 3 treatment methods.
目的：比较毫针、电针和艾灸对膝关节骨关节炎(KOA)大鼠关节软骨形态结构及核转录因子-κB (NF-κB)p65/Nod样受体蛋白3(NLRP3)通路的影响。方法：Wistar大鼠随机分为空白对照组、模型组、毫针组、电针组和艾灸组，每组8只。采用右侧膝关节腔内注射碘乙酸钠复制KOA大鼠模型。3个治疗组均取右侧“犊鼻”“内膝眼”，分别给予毫针、电针和艾条温和灸各15 min，隔日治疗1次，共治疗4周。X线观察各组大鼠右侧膝关节结构并测量直径，透射电镜观察各组大鼠膝关节软骨细胞超微结构，ELISA法检测各组大鼠血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-18含量，Western blot法检测各组大鼠右侧膝关节软骨组织中NF-κB p65、NLRP3、凋亡相关斑点样蛋白(ASC)和TNF-α的表达水平。结果：与空白对照组比较，模型组右侧膝关节间隙变窄严重，软骨细胞明显固缩，线粒体中度肿胀，右侧膝关节直径增加(P<0.01)，血清TNF-α、IL-1β、IL-18含量升高(P<0.01)，膝关节软骨组织中NF-κB p65、NLRP3、TNF-α表达升高(P<0.01)，ASC表达降低(P<0.01)。与模型组比较，3个治疗组右侧膝关节间隙变窄及软骨细胞损伤均得到不同程度改善，右侧膝关节直径缩小(P<0.01，P<0.05)，血清TNF-α含量及膝关节软骨组织NLRP3表达降低(P<0.05，P<0.01)；电针组和艾灸组血清IL-1β、IL-18含量及膝关节软骨组织TNF-α表达降低(P<0.01，P<0.05)，膝关节软骨组织中ASC表达升高(P<0.01)；艾灸组膝关节软骨组织NF-κB p65表达降低(P<0.01)。与电针组比较，艾灸组血清IL-18含量降低(P<0.05)。结论：毫针、电针和艾灸均能减少KOA大鼠膝关节骨赘形成，缓解关节间隙变窄，改善关节软骨超微结构，降低炎性因子水平，调节炎性反应相关NF-κB p65/NLRP3通路，且在3种刺灸方法中艾灸调控作用最明显。.
Keywords: Electroacupuncture; Filiform needling; Inflammatory factor; Knee osteoarthritis; Moxibustion; Nuclear transcription factor-κB p65/Nod-like receptor protein 3 pathway.