Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Traditional Chinese Medicine), Hangzhou 310003, China.
Department of Emergency, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Traditional Chinese Medicine), Hangzhou 310003, China.
Biomed Res Int. 2022 Jun 15;2022:8077058. doi: 10.1155/2022/8077058. eCollection 2022.
Dioscorea nipponica Makino (DNM) is a traditional herb with multiple medicinal functions. This study is aimed at exploring the therapeutic effects of DNM on asthma and the underlying mechanisms involving RKIP-mediated MAPK signaling pathway.
An ovalbumin-induced asthma model was established in mice, which was further administrated with DNM and/or locostatin (RKIP inhibitor). ELISA was performed to detect the serum titers of OVA-IgE and OVA-IgG1, bronchoalveolar lavage fluid (BALF) levels of inflammation-related biomarkers, and tissue levels of oxidative stress-related biomarkers. The expression of RKIP was measured by quantitative real-time PCR, Western blot, immunohistochemistry, and immunofluorescence. HE staining was used to observe the pathological morphology of lung tissues. The protein expression of MAPK pathway-related proteins was detected by Western blot.
Compared with the controls, the model mice exhibited significantly higher serum titers of OVA-IgE and OVA-IgG1, BALF levels of IL-6, IL-8, IL-13, TGF-1, and MCP-1, tissue levels of MDA and ROS, lower BALF levels of IL-10 and IFN-, and tissue level of GSH. DNM relieved the allergic inflammatory response and oxidative stress in the model mice. DNM also recovered the downregulation of RKIP and the pathological injury of lung tissues in asthma mice. In addition, the Raf-1/MEK/MAPK/ERK pathway in the model mice was blocked by DNM. Silencing of RKIP by locostatin weakened the relieving effects of DNM on asthma through activating the Raf-1/MEK/MAPK/ERK pathway.
DNM relieves asthma via blocking the Raf-1/MEK/MAPK/ERK pathway that mediated by RKIP upregulation.
穿龙薯蓣（DNM）是一种具有多种药用功能的传统草药。本研究旨在探讨 DNM 治疗哮喘的疗效及其涉及 RKIP 介导的 MAPK 信号通路的潜在机制。
建立卵白蛋白（OVA）诱导的哮喘小鼠模型，进一步用 DNM 和/或 locostatin（RKIP 抑制剂）处理。通过酶联免疫吸附试验（ELISA）检测血清 OVA-IgE 和 OVA-IgG1 滴度、支气管肺泡灌洗液（BALF）中炎症相关生物标志物水平以及组织中氧化应激相关生物标志物水平。采用实时定量 PCR、Western blot、免疫组化和免疫荧光检测 RKIP 的表达。HE 染色观察肺组织的病理形态。Western blot 检测 MAPK 通路相关蛋白的表达。
与对照组相比，模型组小鼠血清 OVA-IgE 和 OVA-IgG1 滴度、BALF 中 IL-6、IL-8、IL-13、TGF-β1 和 MCP-1 水平、组织中 MDA 和 ROS 水平显著升高，BALF 中 IL-10 和 IFN-γ水平、组织中 GSH 水平显著降低。DNM 缓解了模型小鼠的过敏性炎症反应和氧化应激。DNM 还恢复了哮喘小鼠中 RKIP 的下调和肺组织的病理损伤。此外，DNM 阻断了模型小鼠中的 Raf-1/MEK/MAPK/ERK 通路。用 locostatin 沉默 RKIP 通过激活 Raf-1/MEK/MAPK/ERK 通路削弱了 DNM 对哮喘的缓解作用。
DNM 通过阻断 Raf-1/MEK/MAPK/ERK 通路缓解哮喘，该通路由 RKIP 上调介导。
