Wang Qin, Xu Jiacheng, Li Miaomiao, Chen Yuqing, Xu Yingying, Li Lili, Gong Yongfang, Yang Yan
Department of Pharmacology, School of Basic Medical Sciences, Key Laboratory of Anti-Inflammatory and Immunopharmacology, Ministry of Education, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China.
Naunyn Schmiedebergs Arch Pharmacol. 2024 Mar;397(3):1687-1700. doi: 10.1007/s00210-023-02711-2. Epub 2023 Sep 15.
Fibrotic kidney injury from hepatocarcinogenesis seriously impacts treatment effect. Astragaloside IV (AS-IV), an extract of Astragalus membranaceus, has several pharmacological activities, which are useful in the treatment of edema and fibrosis. Nrf2/HO-1 is a key antioxidant stress pathway and help treatment of kidney injury. Smad3 phosphorylation is implicated in hepatocarcinogenesis. Our previous study clarified that Smad3 is differentially regulated by different phosphorylated forms of Smad3 on hepatocarcinogenesis. Therefore, we investigated the contribution of AS-IV on the therapy of kidney fibrosis from hepatocarcinogenesis. And the focus was on whether the phosphorylation of Smad3 and the regulation of Nrf2/HO-1 pathway were involved during AS-IV therapy and whether there is an effect of Nrf2 knockout on the phosphorylation of Smad3. We performed TGF-β1 stimulation on HK-2 cells and intervened with AS-IV. Furtherly, we investigated renal injury of AS-IV on Nrf2 knockout mice during hepatocarcinogenesis and its mechanism of action. On the one hand, in vitro results showed that AS-IV reduced the ROS and α-SMA expression of HK-2 by promoting the expression pSmad3C/p21 of and Nrf2/HO-1 and suppressed the expression of pSmad3L/PAI-1. On the other hand, the in vivo results of histopathological features, serological biomarkers, and oxidative damage indicators showed that Nrf2 knockout aggravated renal injury. Besides, Nrf2 deletion decreased the nephroprotective effect of AS-IV by suppressing the pSmad3C/p21 pathway and promoting the pSmad3L/PAI-1 pathway. The experimental results were as we suspected. And we identify for the first time that Nrf2 deficiency increases renal fibrosis from hepatocarcinogenesis and attenuates the therapeutic effects of AS-IV via regulating pSmad3C/3L signal pathway.
肝癌发生导致的肾纤维化损伤严重影响治疗效果。黄芪甲苷IV(AS-IV)是黄芪的提取物,具有多种药理活性,对水肿和纤维化的治疗有益。Nrf2/HO-1是关键的抗氧化应激途径,有助于治疗肾损伤。Smad3磷酸化与肝癌发生有关。我们之前的研究阐明,Smad3在肝癌发生过程中受不同磷酸化形式的Smad3差异调节。因此,我们研究了AS-IV对肝癌发生所致肾纤维化治疗的作用。重点在于AS-IV治疗过程中是否涉及Smad3的磷酸化以及Nrf2/HO-1途径的调节,以及Nrf2基因敲除对Smad3磷酸化是否有影响。我们对HK-2细胞进行TGF-β1刺激并用AS-IV进行干预。此外,我们研究了AS-IV对Nrf2基因敲除小鼠在肝癌发生过程中的肾损伤及其作用机制。一方面,体外实验结果表明,AS-IV通过促进pSmad3C/p21和Nrf2/HO-1的表达降低了HK-2细胞的ROS和α-SMA表达,并抑制了pSmad3L/PAI-1的表达。另一方面,组织病理学特征、血清生物标志物和氧化损伤指标的体内实验结果表明,Nrf2基因敲除加重了肾损伤。此外,Nrf2缺失通过抑制pSmad3C/p21途径和促进pSmad3L/PAI-1途径降低了AS-IV的肾保护作用。实验结果正如我们所料。并且我们首次发现,Nrf2缺乏会增加肝癌发生所致的肾纤维化,并通过调节pSmad3C/3L信号通路减弱AS-IV的治疗效果。