Institute of Chinese Medicinal Sciences, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, Guangzhou, 510006, PR China.
Guangdong Province Research Centre for Chinese Integrative Medicine Against Metabolic Disease, PR China; Guangdong TCM Key Laboratory Against Metabolic Diseases, PR China; Key Unit of Modulating Liver to Treat Hyperlipemia State Administration of Traditional Chinese Medicine, PR China; Institute of Chinese Medicinal Sciences, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, Guangzhou, 510006, PR China.
J Ethnopharmacol. 2020 May 10;253:112616. doi: 10.1016/j.jep.2020.112616. Epub 2020 Jan 31.
Renshen Shouwu extract (RSSW) is a patented Traditional Chinese Medicine included in Chinese Pharmacopoeia for neurasthenia, forgetfulness, insomnia, inappetence and excessive fatigue. Our previous study had demonstrated the neuroprotective effect of RSSW against ischemic stroke in rats with middle cerebral artery occlusion (MCAO). However, its underlying mechanism remains unknown.
In this study, we investigated the neurogenesis and angiogenesis effects of RSSW in ischemic stroke rats, and further revealed its underlying mechanism focused on TLR4/NF-κB/NLRP3 signaling pathway.
Firstly, active compounds of RSSW were determined by High Performance Liquid Chromatography (HPLC). Secondly, Middle cerebral artery occlusion (MCAO) was performed to induce ischemic stroke in rats and 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining was employed to evaluate whether MCAO surgery was successfully established. Neurological deficit evaluation was conducted according to the Zea Longa' method. Then, we explored the neurogenesis and angiogenesis effects after oral administration of RSSW (50 mg/kg, 100 mg/kg) in MCAO-induced rats by Immunofluorescence Staining. Moreover, the proteins involved in TLR4/NF-κB/NLRP3 signaling pathway (TLR4, p-NF-κB p65, NF-κB p65, NLRP3, pro-IL-1β, IL-1β, pro-Caspase-1, Caspase-1) were determined by western blotting.
It was observed that RSSW treatment significantly increased the number of newborn neurons and brain microvessel density (MVD) after ischemic stroke. What's more, RSSW treatment significantly downregulated TLR4, p-NF-κB p65/p65, NLRP3, pro-IL-1β, IL-1β, pro-Caspase-1, Caspase-1 proteins involved in TLR4/NF-κB/NLRP3 signaling pathway.
RSSW enhances neurogenesis and angiogenesis via inhibition of TLR4/NF-κB/NLRP3 inflammatory signaling pathway following ischemic stroke in rats. Hence, RSSW may be a promising Chinese Medicine for the treatment of ischemic stroke.
人参首乌提取物（RSSW）是一种专利的中药，被列入中国药典，用于治疗神经衰弱、健忘、失眠、食欲不振和过度疲劳。我们之前的研究表明，RSSW 对大脑中动脉闭塞（MCAO）大鼠缺血性中风具有神经保护作用。然而，其潜在机制尚不清楚。
本研究旨在探讨 RSSW 对缺血性中风大鼠的神经发生和血管生成作用，并进一步揭示其潜在机制，重点关注 TLR4/NF-κB/NLRP3 信号通路。
首先，采用高效液相色谱法（HPLC）测定 RSSW 的活性成分。其次，通过大脑中动脉闭塞（MCAO）诱导大鼠缺血性中风，并采用 2,3,5-三苯基氯化四氮唑（TTC）染色评估 MCAO 手术是否成功。根据 Zea Longa'方法进行神经功能缺损评估。然后，通过免疫荧光染色探讨 RSSW（50mg/kg、100mg/kg）口服给药后对 MCAO 诱导大鼠的神经发生和血管生成作用。此外，通过 Western blot 测定 TLR4/NF-κB/NLRP3 信号通路相关蛋白（TLR4、p-NF-κB p65、NF-κB p65、NLRP3、pro-IL-1β、IL-1β、pro-Caspase-1、Caspase-1）。
结果表明，RSSW 治疗可显著增加缺血性中风后新生神经元数量和脑微血管密度（MVD）。此外，RSSW 治疗可显著下调 TLR4/NF-κB/NLRP3 信号通路相关蛋白（TLR4、p-NF-κB p65/p65、NLRP3、pro-IL-1β、IL-1β、pro-Caspase-1、Caspase-1）。
RSSW 通过抑制 TLR4/NF-κB/NLRP3 炎症信号通路增强缺血性中风后大鼠的神经发生和血管生成。因此，RSSW 可能是一种有前途的治疗缺血性中风的中药。
