School of Pharmacy, Hangzhou Medical College, Hangzhou, People's Republic of China.
School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, People's Republic of China.
Drug Des Devel Ther. 2024 Jul 24;18:3209-3232. doi: 10.2147/DDDT.S462266. eCollection 2024.
Previous studies of our research group have shown that Chuanxiong Renshen Decoction (CRD) has the effect of treating AD, but the exact mechanism of its effect is still not clarified. The aim of this study was to investigate the effect and mechanism of CRD on AD neuroinflammation.
Morris Water Maze (MWM) tests were employed to assess the memory and learning capacity of AD mice. HE and Nissl staining were used to observe the neural cells of mice. The expression of Iba-1 and CD86 were detected by immunohistochemical staining. Utilize UHPLC-MS/MS metabolomics techniques and the KEGG to analyze the metabolic pathways of CRD against AD. Lipopolysaccharide (LPS) induced BV2 microglia cells to construct a neuroinflammatory model. The expression of Iba-1 and CD86 were detected by immunofluorescence and flow cytometry. The contents of TNF-α and IL-1β were detected by ELISA. Western blot assay was used to detect the expression of PPARγ, p-NF-κB p65, NF-κB p65 proteins and inflammatory cytokines iNOS and COX-2 in PPARγ/NF-κB pathway with and without PPARγ inhibitor GW9662.
CRD ameliorated the learning and memory ability of 3×Tg-AD mice, repaired the damaged nerve cells in the hippocampus, reduced the area of Iba-1 and CD86 positive areas in both the hippocampus and cortex regions, as well as attenuated serum levels of IL-1β and TNF-α in mice. CRD-containing serum significantly decreased the expression level of Iba-1, significantly reduced the levels of TNF-α and IL-1β, significantly increased the protein expression of PPARγ, and significantly decreased the proteins expression of iNOS, COX-2 and p-NF-κB p65 in BV2 microglia cells. After addition of PPARγ inhibitor GW9662, the inhibitory effect of CRD-containing serum on NF-κB activation was significantly weakened.
CRD can activate PPARγ, regulating PPARγ/NF-κB signaling pathway, inhibiting microglia over-activation and reducing AD neuroinflammation.
本研究组前期研究表明，川芎人参汤（CRD）具有治疗 AD 的作用，但具体作用机制尚不清楚。本研究旨在探讨 CRD 对 AD 神经炎症的作用及机制。
采用 Morris 水迷宫（MWM）实验评估 AD 小鼠的记忆和学习能力。HE 和尼氏染色观察小鼠神经细胞。免疫组织化学染色检测 Iba-1 和 CD86 的表达。采用 UHPLC-MS/MS 代谢组学技术和 KEGG 分析 CRD 治疗 AD 的代谢途径。脂多糖（LPS）诱导 BV2 小胶质细胞构建神经炎症模型。免疫荧光和流式细胞术检测 Iba-1 和 CD86 的表达。ELISA 检测 TNF-α 和 IL-1β 的含量。Western blot 检测 PPARγ、p-NF-κB p65、NF-κB p65 蛋白及炎症因子 iNOS、COX-2 在 PPARγ/NF-κB 通路中的表达，并用 PPARγ 抑制剂 GW9662 处理。
CRD 改善了 3×Tg-AD 小鼠的学习记忆能力，修复了海马区受损的神经细胞，减少了海马和皮质区 Iba-1 和 CD86 阳性区域的面积，降低了血清中 IL-1β 和 TNF-α 的水平。CRD 含药血清显著降低了 BV2 小胶质细胞中 Iba-1 的表达水平，显著降低了 TNF-α 和 IL-1β 的水平，显著增加了 PPARγ 的蛋白表达，显著降低了 iNOS、COX-2 和 p-NF-κB p65 的蛋白表达。加入 PPARγ 抑制剂 GW9662 后，CRD 含药血清对 NF-κB 激活的抑制作用明显减弱。
CRD 可激活 PPARγ，调控 PPARγ/NF-κB 信号通路，抑制小胶质细胞过度激活，减轻 AD 神经炎症。
