肝损伤的治疗效果在很大程度上依赖于肝脏卓越的再生能力，需要在恢复过程中维持糖/脂质稳态和氧化平衡。黄芪是一种在中国和许多其他国家广泛使用的草药滋补品，被认为具有许多积极作用，包括增强免疫、滋补、抗氧化、保肝、利尿、抗糖尿病、抗癌和祛痰。黄芪被广泛纳入保肝配方中，因为它被认为可以促进肝脏再生。然而，这种保肝作用的确切分子药理学机制仍不清楚。为了探讨黄芪对肝再生的改善作用及其潜在机制，采用70%肝部分切除术（PHx）的小鼠模型来研究黄芪的影响汤剂（HQD）对肝再生的影响。在 PHx 手术前和整个实验过程中，口服 HQD 7 天。 N-乙酰半胱氨酸（NAC）用作肝再生的阳性对照。通过评估肝脏与体重之比（LW/BW）和代表性细胞增殖标记蛋白的表达来评估肝脏再生。使用生化测定、蛋白质印迹、二氢乙锭（DHE）荧光和高碘酸希夫（PAS）染色方法分析氧化应激和葡萄糖代谢。为了了解 AQP9 作为 HQD 的潜在分子靶标在促进肝再生中的作用，采用 td-Tomato 标记的 AQP9 转基因小鼠 (AQP9-RFP) 来确定 AQP9 蛋白的表达模式。采用AQP9基因敲除小鼠（AQP9-/-）评估HQD对AQP9促进肝再生的特异性靶向作用。HQD显着上调野生型（AQP9 / ）小鼠肝脏AQP9表达，减轻肝损伤并促进肝再生。 70% PHx 后。然而，在 AQP9 基因敲除 (AQP9-/-) 小鼠中，HQD 对肝再生的有益影响不存在。此外，HQD 促进肝细胞对甘油的摄取，增强糖异生，同时降低 AQP9 / 但不是 AQP9-/- 小鼠肝脏中的 H2O2 含量和氧化应激水平。此外，黄芪的主要活性物质黄芪甲苷 IV (AS-IV) 和环黄芪醇 (CAG) 可以显着上调 AQP9 的表达并促进 AQP9 / 但不是 AQP9-/- 小鼠的肝再生。 这项研究是第一个证明黄芪及其主要活性成分（AS-IV 和 CAG）通过上调肝细胞中 AQP9 的表达来增加糖异生并减少氧化应激，从而改善肝再生。该研究揭示了黄芪新的分子药理机制，为肝脏疾病提供了一个有前景的治疗靶点。版权所有 © 2023 Elsevier GmbH。版权所有。

The therapeutic efficacy of liver injuries heavily relies on the liver's remarkable regenerative capacity, necessitating the maintenance of glycose/lipids homeostasis and oxidative eustasis during the recovery process. Astragali Radix, an herbal tonic widely used in China and many other countries, is believed to have many positive effects, including immune stimulation, nourishing, antioxidant, liver protection, diuresis, anti-diabetes, anti-cancer and expectorant. Astragali Radix is widely integrated into hepatoprotective formulas as it is believed to facilitate liver regeneration. Nevertheless, the precise molecular pharmacological mechanisms underlying this hepatoprotective effect remain elusive.To investigate the improving effects of Astragali Radix on liver regeneration and the underlying mechanisms.A mouse model of 70% partial hepatectomy (PHx) was employed to investigate the impact of Radix Astragali decoction (HQD) on liver regeneration. HQD was orally administered for 7 days before the PHx procedure and throughout the experiment. N-acetylcysteine (NAC) was used as a positive control for liver regeneration. Liver regeneration was assessed by evaluating the liver-to-body weight ratio (LW/BW) and the expression of representative cell proliferation marker proteins. Oxidative stress and glucose metabolism were analyzed using biochemical assays, Western blotting, dihydroethidium (DHE) fluorescence, and periodic acid-Schiff (PAS) staining methods. To understand the role of AQP9 as a potential molecular target of HQD in promoting liver regeneration, td-Tomato-tagged AQP9 transgenic mice (AQP9-RFP) were employed to determine the expression pattern of AQP9 protein. AQP9 knockout mice (AQP9-/-) were used to assess the specific targeting of AQP9 in the promotion of liver regeneration by HQD.HQD significantly upregulated hepatic AQP9 expression, alleviated liver injury and promoted liver regeneration in wild-type (AQP9+/+) mice after 70% PHx. However, the beneficial impact of HQD on liver regeneration was absent in AQP9 gene knockout (AQP9-/-) mice. Moreover, HQD facilitated the uptake of glycerol by hepatocytes, enhanced gluconeogenesis, and concurrently reduced H2O2 content and oxidative stress levels in AQP9+/+ but not AQP9-/- mouse livers. Additionally, main active substance of Radix Astragali, astragaloside IV (AS-IV) and cycloastragenol (CAG), demonstrated substantial upregulation of AQP9 expression and promoted liver regeneration in AQP9+/+ but not AQP9-/- mice.This study is the first to demonstrate that Radix Astragali and its main active constituents (AS-IV and CAG) improve liver regeneration by upregulating the expression of AQP9 in hepatocytes to increase gluconeogenesis and reduce oxidative stress. The study revealed novel molecular pharmacological mechanisms of Radix Astragali and provided a promising therapeutic target of liver diseases.Copyright © 2023 Elsevier GmbH. All rights reserved.