目的:观察苗医弩药针对膝骨性关节炎（knee osteoarthritis， KOA）大鼠背根神经节（dorsal root ganglion，DRG）电压门控钠通道1.7（Nav1.7）/1.8（Nav1.8）蛋白的影响，探讨弩药针治疗KOA的潜在机制。 方法:将36只雄性SD大鼠随机分为正常组、模型组、弩药针组、皮肤针组。除正常组大鼠（n=9）外，其余采用右膝关节注射碘乙酸钠建立KOA模型。造模结束后，每组随机抽取1只进行模型评估，模型组、弩药针组、皮肤针组每组各8只接受造模处理和干预。弩药针组按照苗医弩药针的操作步骤于大鼠右膝关节面进行干预治疗，皮肤针组采用皮肤针于右膝关节进行局部滚刺。隔日1次，每周3次，连续干预4周。于实验第1、14、21、28、35、42天检测大鼠机械性刺激缩足（von Frey）阈值。干预结束后，通过H&E染色观察大鼠右膝关节软骨组织形态变化；ELISA检测大鼠血清中降钙素基因相关肽（calcitonin gene‑related peptide，CGRP）、P物质（substance P，SP）和前列素E2（prostaglandin E2，PGE2）含量；Western blot和实时荧光定量PCR检测大鼠DRG组织中Nav1.7、Nav1.8蛋白和mRNA相对表达量。 结果:与正常组比，模型组大鼠实验第42天 von Frey 阈值明显降低（P<0.05）；右膝关节软骨表面糜烂缺损，结构层次紊乱，软骨细胞明显减少；血清中CGRP、SP、PGE2含量明显升高（P<0.05）；DRG组织中 Nav1.7、Nav1.8 蛋白和mRNA相对表达量明显升高（P<0.05）。与模型组比较，弩药针组、皮肤针组大鼠实验第42天von Frey阈值明显升高（P<0.05）；右膝关节软骨形态较完整，结构层次较清晰，新生软骨细胞较多；血清中CGRP、SP、PGE2含量明显降低（P<0.05）；DRG组织中Nav1.7、Nav1.8蛋白和mRNA相对表达量明显降低（P<0.05）。与皮肤针组比较，弩药针组大鼠实验第42天von Frey阈值明显升高（P<0.05）；右膝关节软骨表面光滑，结构层次清晰；血清中CGRP、SP、PGE2含量明显降低（P<0.05）；DRG组织中Nav1.7、Nav1.8蛋白和mRNA相对表达量明显降低（P<0.05）。 结论:苗医弩药针可能通过抑制炎性因子CGRP、SP、PGE2的表达和下调DRG中钠通道Nav1.7、Nav1.8蛋白的表达，发挥治疗KOA作用。

Objective:To observe the effects of Miao medicinal crossbow acupuncture on voltage‑gated sodium channel 1.7 （Nav1.7）/voltage‑gated sodium channel 1.8 （Nav1.8） proteins in the dorsal root ganglion （DRG） of rats with knee osteoarthritis （KOA）， and to explore the potential mechanism of the crossbow acupuncture treating KOA. Methods:A total of 36 rats were randomly divided into the normal group， the model group， the Miao medicinal crossbow acupuncture group， and the dermal needle group. Except for the normal group （n=9）， the rest of the rats were injected with sodium iodoacetate into the right knee joint to establish KOA models. One rat was randomly selected from each group for model evaluation after modeling， and there were 8 rats in each group of the model group， the Miao medicinal crossbow acupuncture group， and the dermal needle group that received modeling procedure and intervention. The rats in the Miao medicinal crossbow acupuncture group were treated with Miao medicinal crossbow acupuncture following the operation procedures， and the rats in the dermal needle group were treated with rolling a skin needle on the right knee joint surface. The intervention was given once every other day， 3 times a week for 4 weeks. On day 1， 14， 21， 28， 35 and 42 of the experiment， paw withdrawal （von Frey） threshold was measured. After the intervention， H&E staining was used to observe the morphological changes of cartilage tissue in the right knee joint of rats. The contents of calcitonin gene‑related peptide （CGRP）， substance P （SP）， and prostaglandin E2 （PGE2） in serum of rats were detected by ELISA. The relative expressions of Nav1.7 and Nav1.8 proteins and mRNA in DRG tissues of rats were detected by Western blot and real‑time fluorescence quantitative PCR. Results:Compared to the normal group， von Frey threshold was significantly decreased in the model group on day 42 （P<0.05）. The cartilage surface of the right knee joint was eroded and defective， with structural disorders and a significant reduction in chondrocytes. The levels of CGRP， SP， and PGE2 in serum were significantly increased （P<0.05）. The relative expressions of Nav1.7 and Nav1.8 proteins and mRNA in DRG tissue were significantly increased （P<0.05）. Compared to the model group， von Frey threshold was significantly increased on the day 42 of experiment in the Miao medicinal crossbow acupuncture group and the dermal needle group （P<0.05）. The cartilage of the right knee joint was more complete in morphology， with clearer structural layers and more neoplastic chondrocytes. The levels of CGRP， SP， and PGE2 in serum were significantly decreased （P<0.05）. The relative expressions of Nav1.7 and Nav1.8 proteins and mRNA in DRG tissue were significantly decreased （P<0.05）. Compared to the dermal needle group， von Frey threshold was significantly increased on the day 42 of experiment in the Miao medicinal crossbow acupuncture group （P<0.05）. The cartilage surface of the right knee joint was smooth and the structural layers were clear. The levels of CGRP， SP， and PGE2 in serum were significantly decreased （P<0.05）. The relative expressions of Nav1.7 and Nav1.8 proteins and mRNA in DRG tissue were significantly decreased （P<0.05）. Conclusion:Miao medicinal crossbow acupuncture may exert its therapeutic effects on KOA by inhibiting the expression of inflammatory factors such as CGRP， SP， and PGE2， as well as downregulating the expression of sodium channels Nav1.7 and Nav1.8 proteins in the DRG.

R29

贵州省科技计划项目 ［黔科合支撑（2023）一般240］