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目的：观察针刺“足三里”对弗氏完全佐剂（Complete Freund′s Adjuvant,CFA）诱导关节炎小鼠足爪部位巨噬细胞募集及趋化因子表达的影响，探讨针刺改善CFA小鼠病变部位炎症反应的可能机制。方法：30只雄性BALB/c小鼠随机分为盐水组、模型组、针刺组，每组10只。采用右足跖皮下注射CFA建立关节炎模型。其中针刺组给予双侧“足三里”针刺30 min，第1～7天，每天1次，第8～15天，隔天1次。期间观察各组小鼠热辐射痛阈值、足趾容积及关节炎症评分。采用流式细胞术检测足爪部位巨噬细胞数量、M1型和M2型巨噬细胞的数量；采用RT-qPCR检测足爪部位促炎因子IL-1β、TNF-α，抗炎因子IL-10、IL-4在基因水平的表达；采用液相芯片技术检测足爪部位抗炎因子IL-10、IL-4及趋化因子CCL2、CCL7、CCL12、CX3CL1在蛋白水平的表达。结果：与盐水组相比，造模后模型组热辐射痛阈阈值显著降低（P<0.01），足趾容积、关节炎症评分均显著升高（P<0.01）；模型组足爪部位巨噬细胞数量及M1、M2型巨噬细胞均显著升高（P<0.01）；模型组足爪部位TNF-α、IL-1β在基因水平的表达升高（P<0.01）,IL-10、IL-4在基因水平的表达无明显变化（P>0.05）；模型组足爪部位IL-10、IL-4在蛋白水平的表达显著升高（P<0.01），趋化因子CCL2、CCL7、CCL12、CX3CL1均显著升高（P<0.01）。。与模型组相比，针刺组热辐射痛阈值显著升高（P<0.01），足趾容积、关节炎症评分均降低（P<0.05）；针刺组足爪部位巨噬细胞数量及M1、M2型巨噬细胞均显著降低（P<0.01）；针刺组足爪部位TNF-α、IL-1β在基因水平的表达降低（P<0.01,P<0.05）,IL-10、IL-4在基因水平的表达无明显变化（P>0.05）；针刺组小鼠足爪部位IL-10在蛋白水平的表达升高（P<0.05）,IL-4表达在蛋白水平无明显变化（P>0.05），趋化因子CCL2、CCL7、CCL12、CX3CL1表达均降低（P<0.05）。结论：针刺“足三里”能有效减轻CFA小鼠关节炎症反应，可能是通过减少病变局部趋化因子从而减轻巨噬细胞募集来实现的。
Abstract：Objective: To observe the effect of acupuncture at “Zusanli”(ST36) on macrophage recruitment and chemokine expression in the paw of arthritis mice induced by Complete Freund's Adjuvant(CFA), and to explore the possible mechanism of acupuncture improving the inflammatory response in the lesion site of CFA mice. Methods: A total of 30 male BALB/c mice were randomly divided into the saline group, model group, and acupuncture group, with 10 mice in each group. The arthritis model was established by subcutaneous injection of CFA into the right foot. The acupuncture group was given bilateral “Zusanli” acupuncture for 30 min, once a day from day 1 to day 7, and once every other day from day 8 to day 15. The thermal radiation pain threshold, toe volume and joint inflammation score of mice in each group were observed during the period. Flow cytometry was used to detect the number of macrophages, M1 and M2 macrophages in the paw. RT-qPCR was used to detect the expression of pro-inflammatory factors IL-1β and TNF-α, anti-inflammatory factors IL-10 and IL-4 at the gene level. The expression of anti-inflammatory factors IL-10, IL-4 and chemokines CCL2, CCL7, CCL12, and CX3CL1 at the protein level was detected by liquid chip technology. Results: Compared to the saline group, the thermal radiation pain threshold of the model group was significantly decreased(P<0.01), and the toe volume and joint inflammation score were significantly increased(P<0.01). The number of macrophages and M1 and M2 macrophages in the paw of the model group were significantly increased(P<0.01). The expression of TNF-α and IL-1β in the paw of the model group increased at the gene level(P<0.01), and the expression of IL-10 and IL-4 at the gene level did not change significantly(P>0.05). The expression of IL-10 and IL-4 in the paw of the model group was significantly increased at the protein level(P<0.01), and the chemokines CCL2, CCL7, CCL12, and CX3CL1 were significantly increased(P<0.01).Compared to the model group, the thermal radiation pain threshold of the acupuncture group was significantly increased(P<0.01), and the toe volume and joint inflammation score were decreased(P<0.05). The number of macrophages and M1 and M2macrophages in the foot claws of the acupuncture group were significantly decreased(P<0.01). The expression of TNF-α and IL-1β in the foot claws of the acupuncture group decreased at the gene level(P<0.01, P<0.05), and the expression of IL-10 and IL-4 at the gene level did not change significantly(P>0.05). The expression of IL-10 in the foot paw of the acupuncture group was increased at the protein level(P<0.05), and the expression of chemokines CCL2, CCL7, CCL12, and CX3CL1 was decreased(P<0.05). Conclusion: Acupuncture at “Zusanli”can effectively reduce the joint inflammatory response in CFA mice, which may be achieved by reducing the recruitment of macrophages by reducing the chemokines in the lesion.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20240830.002
中图分类号:R245
引用信息:
[1]殷建朝,王悦,公一囡,等.针刺“足三里”对弗氏完全佐剂诱导关节炎小鼠足爪部位巨噬细胞募集及趋化因子表达的影响[J].海南医科大学学报,2025,31(09):683-690.DOI:10.13210/j.cnki.jhmu.20240830.002.
基金信息:
国家自然科学基金青年科学基金项目（82004473）~~
2024-08-31
2024-08-31
2024-08-31