This study aims to investigate the hepatoprotective effect of the antipsychotic drug trifluoperazine (TFP) against cyclophosphamide (CPA)-induced hepatic injury by exploring its effect on autophagy and the Nrf2/HO-1 signaling pathway. The hepatotoxicity of CPA was assessed by biochemical analysis of the serum hepatotoxicity markers (ALT, AST, and direct bilirubin), histopathological examination, and ultrastructure analysis by transmission electron microscopy (TEM). The ELISA technique was used to assess the hepatic content of oxidative stress (MDA and SOD) and inflammatory markers (IL-1β and TNF-α). Immunohistochemical assessment was used to investigate the hepatic expression of NF-κB, Nrf2, caspase-3, as well as autophagy flux markers (p62 and LC3B). The mRNA expression of HO-1 was assessed using RT-qPCR. Western blot assay was used to determine the expression of p-AKT and p-mTOR. TFP improved CPA-induced hepatotoxicity by reducing the elevated hepatotoxicity markers, and alleviating the histopathological changes with improving ultrastructure alterations. It also reduced oxidative stress by reducing MDA content and upregulating SOD activity. In addition, it exhibited anti-inflammatory and anti-apoptotic effects by decreasing NF-κB expression, IL-1β, TNF-α levels, and caspase-3 expression. Furthermore, TFP-induced hepatoprotection was mediated by favoring Nrf2 expression and increasing the mRNA level of HO-1. As well, it improved autophagy by increasing LC3B expression concurrently with reducing p62 expression. Moreover, TFP modulated the AKT/mTOR pathway by reducing the expression of p-AKT and p-mTOR. TFP significantly protected against CPA-induced hepatotoxicity by upregulating Nrf2/HO-1 signaling along with enhancement of protective autophagy via inhibition of the AKT/mTOR signaling pathway.

中文翻译：

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Trifluoperazine 通过调节 AKT/mTOR 驱动的自噬和 Nrf2/HO-1 信号级联减轻环磷酰胺诱导的小鼠肝脏氧化应激、炎症和细胞凋亡

本研究旨在通过探讨抗精神病药物三氟拉嗪（TFP）对自噬和Nrf2/HO-1信号通路的影响，探讨其对环磷酰胺（CPA）诱导的肝损伤的保肝作用。通过血清肝毒性标志物（ALT、AST和直接胆红素）的生化分析、组织病理学检查和透射电子显微镜（TEM）超微结构分析来评估CPA的肝毒性。采用ELISA技术评估肝脏氧化应激（MDA和SOD）和炎症标志物（IL-1β和TNF-α）的含量。使用免疫组织化学评估来研究 NF-κB、Nrf2、caspase-3 以及自噬流标记物（p62 和 LC3B）的肝脏表达。使用 RT-qPCR 评估 HO-1 的 mRNA 表达。 Western blot法检测p-AKT和p-mTOR的表达。 TFP 通过降低升高的肝毒性标志物来改善 CPA 诱导的肝毒性，并通过改善超微结构改变来减轻组织病理学变化。它还通过降低 MDA 含量和上调 SOD 活性来减少氧化应激。此外，它还通过降低 NF-κB 表达、IL-1β、TNF-α 水平和 caspase-3 表达来发挥抗炎和抗凋亡作用。此外，TFP 诱导的肝保护作用是通过促进 Nrf2 表达和增加 HO-1 mRNA 水平介导的。此外，它还通过增加 LC3B 表达同时减少 p62 表达来改善自噬。此外，TFP 通过降低 p-AKT 和 p-mTOR 的表达来调节 AKT/mTOR 通路。 TFP 通过上调 Nrf2/HO-1 信号传导以及通过抑制 AKT/mTOR 信号通路增强保护性自噬来显着防止 CPA 诱导的肝毒性。