Su Yunfang, Liu Ningning, Sun Ruiqin, Ma Jinlian, Li Zhonghua, Wang Pan, Ma Huifen, Sun Yiran, Song Junying, Zhang Zhenqiang
Henan Engineering Research Center for Prevention and Treatment of Major Chronic Diseases with Chinese Medicine, Academy of Chinese Medical Sciences, Henan University of Chinese Medicine, Zhengzhou, China.
The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, China.
Front Pharmacol. 2023 Feb 8;14:1115387. doi: 10.3389/fphar.2023.1115387. eCollection 2023.
Radix Rehmanniae Praeparata (RRP, Shu Dihuang in Cinese) is widely used as primal medicine in Chinese herbal formula for the treatment of Alzheimer's disease (AD). However, the underlying mechanism of RRP for AD remains unclear. The aim of this study was to investigate the therapeutic effect of RRP on intracerebroventricular injection of streptozotocin (ICV-STZ)-induced AD model mice and its potential mechanism. ICV-STZ mice were continuously gavaged with RRP for 21 days. The pharmacological effects of RRP were evaluated by behavioral tests, brain tissue H&E staining and hippocampal tau protein phosphorylation levels. The expression levels of insulin receptor (INSR), IRS-1, pSer473-AKT/AKT and pSer9-GSK-3β/GSK-3β proteins in hippocampal and cortical tissues were detected by Western-blot method. The 16S rRNA gene sequencing was used to analyze the changes of intestinal microbiota in mice. The compounds in RRP were analyzed by mass spectrometry and their binding ability to INSR proteins was detected by molecular docking. The results showed that RRP ameliorated cognitive dysfunction and neuronal pathological changes of brain tissue in ICV-STZ mice, reduced tau protein hyperphosphorylation, INSR, IRS-1, pSer473-AKT/AKT, and pSer9-GSK-3β/GSK-3β levels in hippocampal and cortical tissues. Meanwhile, RRP reversed ICV-STZ-induced dysregulation of intestinal microbiota in AD mice. Mass spectrometry analysis showed that the RRP consisted mainly of seven compounds, namely Acteoside (Verbascoside), 5-Hydroxymethyl-2-furaldehyde (5-HMF), Apigenin7-O-glucuronide, Icariin, Gallic acid, Quercetin-3β-D-glucoside, and Geniposide. Molecular docking results further indicated that the compounds in RRP have binding ability to INSR protein and potential multiple synergistic effects. RRP ameliorates cognitive dysfunction and brain histopathological changes in AD mice. The mechanism of RRP ameliorating AD may be related to the regulation of INSR/IRS-1/AKT/GSK-3β signaling pathway and intestinal microbiota. This study supports the potential anti-AD efficacy of RRP and initially reveals the pharmacological mechanism of RRP, providing a theoretical basis for further clinical application of RRP.
熟地黄在中国草药配方中被广泛用作治疗阿尔茨海默病（AD）的主要药物。然而，熟地黄治疗AD的潜在机制仍不清楚。本研究旨在探讨熟地黄对脑室内注射链脲佐菌素（ICV-STZ）诱导的AD模型小鼠的治疗作用及其潜在机制。对ICV-STZ小鼠连续灌胃熟地黄21天。通过行为测试、脑组织苏木精-伊红染色和海马tau蛋白磷酸化水平评估熟地黄的药理作用。采用蛋白质免疫印迹法检测海马和皮质组织中胰岛素受体（INSR）、胰岛素受体底物-1（IRS-1）、磷酸化丝氨酸473的蛋白激酶B（pSer473-AKT）/蛋白激酶B（AKT）和磷酸化丝氨酸9的糖原合成酶激酶-3β（pSer9-GSK-3β）/糖原合成酶激酶-3β（GSK-3β）蛋白的表达水平。利用16S rRNA基因测序分析小鼠肠道微生物群的变化。通过质谱分析熟地黄中的化合物，并通过分子对接检测其与INSR蛋白的结合能力。结果表明，熟地黄改善了ICV-STZ小鼠的认知功能障碍和脑组织的神经元病理变化，降低了海马和皮质组织中tau蛋白的过度磷酸化、INSR、IRS-1、pSer473-AKT/AKT和pSer9-GSK-3β/GSK-3β的水平。同时，熟地黄逆转了ICV-STZ诱导的AD小鼠肠道微生物群失调。质谱分析表明，熟地黄主要由七种化合物组成，即毛蕊花糖苷、5-羟甲基糠醛、芹菜素-7-O-葡萄糖醛酸苷、淫羊藿苷、没食子酸、槲皮素-3-β-D-葡萄糖苷和栀子苷。分子对接结果进一步表明，熟地黄中的化合物具有与INSR蛋白结合的能力和潜在的多重协同作用。熟地黄改善了AD小鼠的认知功能障碍和脑组织病理变化。熟地黄改善AD的机制可能与调节INSR/IRS-1/AKT/GSK-3β信号通路和肠道微生物群有关。本研究支持熟地黄潜在的抗AD疗效，并初步揭示了熟地黄的药理机制，为熟地黄进一步的临床应用提供了理论依据。
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