Medical College, Inner Mongolia University for Nationalities, Tongliao, 028000, Inner Mongolia, PR China.
Affiliated Hospitals, Inner Mongolia University for Nationalities, Tongliao, 028000, Inner Mongolia, PR China.
J Ethnopharmacol. 2021 Jun 12;273:114002. doi: 10.1016/j.jep.2021.114002. Epub 2021 Mar 8.
The traditional Chinese medicine Sanweidoukou decoction (DK-3) was a classical formula for the treatment of nervous system diseases, recorded in the Chinese medical classic Sibu Yidian.
The present study is aim to investigate the neuroprotective effects of DK-3 on β-amyloid (Aβ) protein -induced AD-like pathologies and underlying molecular mechanisms both in vitro and in vivo studies.
Hydrolysates of DK-3 were analyzed by LC-ESI-MS/MS. In vitro, MTT was utilized to examine effects of DK-3 on Aβ-induced cytotoxicity in PC12 cells. In vivo, male Sprague-Dawley rats were administered with Aβ to induce AD-like pathologies and behavioral evaluations were conducted via Morris water maze (MWM) test. Histopathological changes were observed by Hematoxylin-eosin (HE) straining. Immunohistochemistry (IHC) was used to detect the tau hyperphosphorylation at Thr181 site. The expression levels of tau hyperphosphorylation, inflammation-related cytokines such as COX-2, iNOS, TNF-α, IL-1β, IL-6, the phosphorylated state of various mitogen-activated protein kinase (MAPK) signaling molecules (p38 MAPK, ERK, and JNK) and activation of nuclear factor κB (NF-κB) in vitro and in vivo were assessed via Western blot.
In vitro, DK-3 dose-dependently increased cell viability of PC12 cells induced by Aβ. In vivo, DK-3 improved learning and memory abilities of Aβ-induced AD-like rats. Moreover, DK-3 reversed hyperphosphorylation of tau and reduced the production of inflammation-related cytokines through significantly inhibited MAPK and NF-κB signaling pathways both in vitro and in vivo studies.
The present study suggested that the traditional Chinese medicine DK-3 may play a role in preventing and treating AD by reducing the hyperphosphorylation of tau protein and the expressions of inflammation-related cytokines via modulating the MAPK/NF-κB signaling pathways.
中药三联豆口汤（DK-3）是一种治疗神经系统疾病的经典方剂，记录在中国医学经典《四补医典》中。
本研究旨在通过体外和体内研究，探讨 DK-3 对β-淀粉样蛋白（Aβ）蛋白诱导的 AD 样病变的神经保护作用及其潜在的分子机制。
通过 LC-ESI-MS/MS 分析 DK-3 的水解产物。在体外，MTT 用于检测 DK-3 对 PC12 细胞中 Aβ诱导的细胞毒性的影响。在体内，雄性 Sprague-Dawley 大鼠给予 Aβ诱导 AD 样病变，并通过 Morris 水迷宫（MWM）测试进行行为评估。苏木精-伊红（HE）染色观察组织病理学变化。免疫组织化学（IHC）用于检测 Thr181 位点 tau 过度磷酸化。通过 Western blot 评估体外和体内 tau 过度磷酸化、炎症相关细胞因子（如 COX-2、iNOS、TNF-α、IL-1β、IL-6）、各种丝裂原活化蛋白激酶（MAPK）信号分子（p38 MAPK、ERK 和 JNK）的磷酸化状态以及核因子 κB（NF-κB）的激活。
在体外，DK-3 剂量依赖性地增加 Aβ诱导的 PC12 细胞活力。在体内，DK-3 改善了 Aβ诱导的 AD 样大鼠的学习和记忆能力。此外，DK-3 通过显著抑制 MAPK 和 NF-κB 信号通路，在体外和体内研究中均逆转了 tau 的过度磷酸化并减少了炎症相关细胞因子的产生。
本研究表明，中药 DK-3 通过调节 MAPK/NF-κB 信号通路，减少 tau 蛋白的过度磷酸化和炎症相关细胞因子的表达，可能在预防和治疗 AD 中发挥作用。
