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基于Wnt/β-catenin信号通路的穴位敷贴联合针刺对强直性脊柱炎模型大鼠的改善作用

Efficacy of acupoint application combined with acupuncture in improving ankylosing spondylitis of model rats based on the Wnt/β-catenin signaling pathway

    摘要
  • 摘要:
    目的 基于Wnt/β-catenin信号通路探讨中药穴位敷贴联合针刺对强直性脊柱炎(AS)模型大鼠的改善作用。
    方法 将60只Wistar大鼠随机分为空白对照组、模型组、阳性对照组(柳氮磺吡啶)、中药穴位敷贴组、针刺组、中药穴位敷贴联合针刺组，每组10只。评估各组大鼠关节炎症状评分; 采用苏木精-伊红(HE)染色检测骶髂关节组织的病理学损伤; 采用酶联免疫吸附测定(ELISA)检测血清循环免疫复合物(CIC)和干扰素γ(IFN-γ)水平，采用碱性磷酸酶(ALP)试剂盒检测血清ALP水平; 采用Western blot检测骶髂关节组织中Osterix、Runt相关转录因子2(Runx2)、Wnt家族成员3A(Wnt3a)、低密度脂蛋白受体相关蛋白5(LRP5)、糖原合酶激酶3β(GSK3β)、β-catenin蛋白的表达。
    结果 相较于空白对照组，模型组骶髂关节组织呈现显著的病理学损伤，血清CIC、IFN-γ、ALP水平升高，骶髂关节组织中Osterix、Runx2、Wnt3a、LRP5、β-catenin蛋白表达上调，骶髂关节组织中GSK3β蛋白表达下调，差异均有统计学意义(P < 0.05)。相较于模型组，阳性对照组、中药穴位敷贴组、针刺组、中药穴位敷贴联合针刺组骶髂关节组织病理学损伤减轻，血清CIC、IFN-γ、ALP水平降低，骶髂关节组织中Osterix、Runx2、Wnt3a、LRP5、β-catenin蛋白表达下调，骶髂关节组织中GSK3β蛋白表达上调，差异有统计学意义(P < 0.05)。相较于阳性对照组、中药穴位敷贴组、针刺组，中药穴位敷贴联合针刺组骶髂关节组织病理学损伤减轻，血清CIC、IFN-γ、ALP水平降低，骶髂关节组织中Osterix、Runx2、Wnt3a、LRP5、β-catenin蛋白表达下调，骶髂关节组织中GSK3β蛋白表达上调，差异有统计学意义(P < 0.05)。
    结论 中药穴位敷贴联合针刺能够抑制AS大鼠异位骨化、炎症反应，其机制可能与抑制Wnt/β-catenin信号通路相关。

     

    Abstract:
    Objective To investigate the efficacy of traditional Chinese medicine (TCM) acupoint application combined with acupuncture in improving ankylosing spondylitis (AS) of model rats based on the Wnt/β-catenin signaling pathway.
    Methods Sixty Wistar rats were randomly divided into blank control group, model group, positive control group (sulfasalazine), TCM acupoint application group, acupuncture group, and TCM acupoint application combined with acupuncture group, with 10 rats in each group. The arthritis symptom score of rats in each group was evaluated. Hematoxylin-eosin (HE) staining was used to detect the pathological damage of sacroiliac joint tissues. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum circulating immune complexes (CIC) and interferon-γ (IFN-γ), and an alkaline phosphatase (ALP) kit was used to detect the serum ALP level. Western blot was used to detect the expression levels of Osterix, Runt-related transcription factor 2 (Runx2), Wnt family member 3A (Wnt3a), low-density lipoprotein receptor-related protein 5 (LRP5), glycogen synthase kinase 3β (GSK3β), and β-catenin proteins in sacroiliac joint tissues.
    Results Compared with the blank control group, the model group showed significant pathological damage in sacroiliac joint tissues, increased serum levels of CIC, IFN-γ and ALP, upregulated expressions of Osterix, Runx2, Wnt3a, LRP5, and β-catenin proteins, and downregulated expression of GSK3β protein in sacroiliac joint tissues (P < 0.05). Compared with the model group, the positive control group, TCM acupoint application group, acupuncture group, and TCM acupoint application combined with acupuncture group showed reduced pathological damage in sacroiliac joint tissues, decreased serum levels of CIC, IFN-γ and ALP, downregulated expressions of Osterix, Runx2, Wnt3a, LRP5, and β-catenin proteins, and upregulated expression of GSK3β protein in sacroiliac joint tissues, with statistically significant between-group differences (P < 0.05). Compared with the positive control group, TCM acupoint application group, and acupuncture group, the TCM acupoint application combined with acupuncture group showed reduced pathological damage, decreased serum levels of CIC, IFN-γ and ALP, downregulated expressions of Osterix, Runx2, Wnt3a, LRP5, and β-catenin proteins, and upregulated expression of GSK3β protein in sacroiliac joint tissues, with statistically significant between-group differences (P < 0.05).
    Conclusion TCM acupoint application combined with acupuncture can inhibit ectopic ossification and inflammatory response in AS rats, and its mechanism may be related to the inhibition of the Wnt/β-catenin signaling pathway.

     

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