Hou Lili, Zhu Lei, Zhang Min, Zhang Xingyi, Zhang Guoqing, Liu Zhenwei, Li Qiang, Zhou Xin
Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai, China.
Department of Respiratory Medicine, Zhongshan Hospital, Fudan University, Shanghai, China.
Cell Physiol Biochem. 2017;41(6):2230-2241. doi: 10.1159/000475638. Epub 2017 Apr 25.
Present study was performed to examine whether ADH was implicated in psychological stress asthma and to explore the underlying molecular mechanism.
We not only examined ADH levels in the cerebrospinal fluid (CSF) via radioimmunoassay, but also measured ADH receptor (ADHR) expression in airway-related vagal preganglionic neurons (AVPNs) through real-time PCR in all experimental mice. Western blotting was performed to evaluate the relationship between ADH and PKA/PKC in psychological stress asthma. Finally, the role of PKA/PKC in psychological stress asthma was analyzed.
Marked asthma exacerbations were noted owing to significantly elevated levels of ADH and ADHR after psychological stress induction as compared to OVA alone (asthma group). ADHR antagonists (SR-49095 or SR-121463A) dramatically lowered higher protein levels of PKAα and PKCα induced by psychological stress as compared to OVA alone, suggesting the correlation between ADH and PKA/PKC in psychological stress asthma. KT-5720 (PKA inhibitor) and Go-7874 (PKC inhibitor) further directly revealed the involvement of PKA/PKC in psychological stress asthma. Some notable changes were also noted after employing PKA and PKC inhibitors in psychological stress asthma, including reduced asthmatic inflammation (lower eosinophil peroxidase (EPO) activity, myeloperoxidase (MPO) activity, immunoglobulin E (IgE) level, and histamine release), substantial decrements in inflammatory cell counts (eosinophils and lymphocytes), and decreased cytokine secretion (IL-6, IL-10, and IFN-γ), indicating the involvement of PKA/PKC in asthma exacerbations induced by psychological stress.
Our results strongly suggested that ADH participated in psychological stress-induced asthma exacerbations via PKA/PKC signal pathway in AVPNs.
进行本研究以检验抗利尿激素（ADH）是否与心理应激性哮喘有关，并探索其潜在的分子机制。
我们不仅通过放射免疫分析法检测了所有实验小鼠脑脊液（CSF）中的ADH水平，还通过实时聚合酶链反应（PCR）测量了气道相关迷走神经节前神经元（AVPNs）中ADH受体（ADHR）的表达。采用蛋白质免疫印迹法评估心理应激性哮喘中ADH与蛋白激酶A（PKA）/蛋白激酶C（PKC）之间的关系。最后，分析了PKA/PKC在心理应激性哮喘中的作用。
与仅使用卵清蛋白（OVA）的情况（哮喘组）相比，心理应激诱导后ADH和ADHR水平显著升高，导致哮喘明显加重。与仅使用OVA相比，ADHR拮抗剂（SR-49095或SR-121463A）显著降低了心理应激诱导的PKAα和PKCα较高的蛋白水平，提示心理应激性哮喘中ADH与PKA/PKC之间存在相关性。KT-5720（PKA抑制剂）和Go-7874（PKC抑制剂）进一步直接揭示了PKA/PKC参与心理应激性哮喘。在心理应激性哮喘中使用PKA和PKC抑制剂后也观察到一些显著变化，包括哮喘炎症减轻（嗜酸性粒细胞过氧化物酶（EPO）活性、髓过氧化物酶（MPO）活性、免疫球蛋白E（IgE）水平和组胺释放降低）、炎症细胞计数（嗜酸性粒细胞和淋巴细胞）大幅减少以及细胞因子分泌（白细胞介素-6（IL-6）、白细胞介素-10（IL-10）和干扰素-γ（IFN-γ））减少，表明PKA/PKC参与心理应激诱导的哮喘加重。
我们的结果强烈提示ADH通过AVPNs中的PKA/PKC信号通路参与心理应激诱导的哮喘加重。
