降气平哮方调控CLCA1/Ca2+/SNARE信号改善哮喘气道黏液高分泌的机制研究

Mechanistic Study of Jiangqi Pingxiao Formula in Ameliorating Mucus Hypersecretion in Asthmatic Airways via Regulating CLCA1/Ca2+/SNARE Pathway

    摘要
  • 摘要:
    目的 探讨降气平哮方调控Ca2+与膜融合过程对哮喘气道黏液高分泌的作用机制。
    方法 采用卵清蛋白致敏小鼠建立哮喘模型，设正常组，模型组，降气平哮方低（10.0 g·kg-1）、中（20.0 g·kg-1）、高（30.0 g·kg-1）剂量组，地塞米松组（1.0 mg·kg-1）和盐酸氨溴索组（8.0 mg·kg-1）。在造模期间通过行为学观察小鼠状态、HE染色和PAS染色评估肺部病理状态，ELISA法检测小鼠肺部炎症因子白细胞介素（IL）-4与IL-13水平，qPCR检测黏蛋白mRNA表达，比色法检测小鼠肺组织中Ca2+浓度，免疫组织化学和Western blot法检测钙激活氯通道调节因子1（CLCA1）和膜融合相关蛋白的表达。
    结果 与正常组相比，模型组气道炎症评分，黏液分泌，IL-13、IL-4水平，Ca2+浓度及CLCA1、突触体相关蛋白23（SNAP23）、可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体（SNARE）、突触融合蛋白（STX3）、囊泡相关膜蛋白（VAMP）8和VAMP2的表达均显著升高（P < 0.05，P < 0.01，P < 0.001）；与模型组相比，降气平哮方可有效改善哮喘小鼠的一般状况，显著减轻小鼠气道炎症和黏液分泌（P < 0.05，P < 0.001）；降低肺组织中IL-4、IL-13水平（P < 0.001）；抑制黏蛋白基因的过表达（P < 0.05，P < 0.01，P < 0.001）；减少细胞内Ca2+浓度（P < 0.05，P < 0.001）；下调CLCA1及SNAP23、SNARE、STX3、VAMP8、VAMP2等膜融合通路关键蛋白表达。
    结论 降气平哮方可能通过抑制CLCA1/Ca2+/SNARE信号减轻哮喘气道黏液高分泌，为降气平哮方治疗提供了新的分子机制靶点。

     

    Abstract:
    OBJECTIVE To investigate the mechanism by which Jiangqi Pingxiao Formula (JQPXF) regulates Ca2+and membrane fusion to attenuate airway mucus hypersecretion in asthma.
    METHODS An ovalbumin-induced asthma model was established in mice. The experimental groups included the normal control group, the asthma model group, JQPXF low- (10.0 g·kg-1), medium- (20.0 g·kg-1), and high-dose (30.0 g·kg-1) groups, the dexamethasone group (1.0 mg·kg-1), and the ambroxol hydrochloride group (8.0 mg·kg-1). During the model construction period, the status of mice were observed through behavioral observation, and lung pathological status were evaluated by HE and PAS staining. The levels of inflammatory cytokines interleukin (IL)-4 and IL-13 in mouse lungs were detected using the ELISA method. qPCR was used to detect mucin mRNA expression, calcium ion (Ca2+) concentration in mouse lung tissue was measured by colorimetric assays, and immunohistochemistry and Western blot analyses were used to assess the expression of calcium-activated chloride channel modulator 1 (CLCA1) and membrane fusion-related proteins.
    RESULTS Compared with the normal group, the model group exhibited significantly increased airway inflammation scores, mucus secretion, IL-13 and IL-4 levels, calcium ion concentration, and expression of CLCA1; synaptophysin-associated protein 23 (SNAP23), soluble N-ethylmaleimide-sensitive receptor protein (SNARE), synaptotagmin (STX3), vesicle-associated membrane protein 8 (VAMP8), and vesicle-associated membrane protein 2 (VAMP2) were significantly elevated (P < 0.05, P < 0.01, P < 0.001). Compared with the model group, JQPXF effectively improved various clinical signs in asthmatic mice, significantly alleviating airway inflammation and mucus secretion (P < 0.05, P < 0.001); reduced IL-4 and IL-13 levels in lung tissue (P < 0.001); suppressed the overexpression of mucin genes (P < 0.05, P < 0.01, P < 0.001); decreased intracellular Ca2+ concentration (P < 0.05, P < 0.001); and downregulated the expression of key proteins in the membrane fusion pathway, including CLCA1, SNAP23, SNARE, STX3, VAMP8 and VAMP2.
    CONCLUSION JQPXF may ameliorate mucus hypersecretion in asthmatic airways by suppressing the CLCA1/Ca2+/SNARE signaling pathway, providing a novel molecular mechanism target for JQPXF therapy.

     

    • HTML全文
    • 参考文献(35)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回