School of Medical Technology, Xinxiang Medical University, NO. 601 Jinsui Avenue, Xinxiang, Henan 453003, China.
School of Pharmacy, Xinxiang Medical University, Xinxiang, China.
Phytomedicine. 2024 Nov;134:155956. doi: 10.1016/j.phymed.2024.155956. Epub 2024 Aug 15.
Alcoholic liver disease (ALD) significantly contributes to global liver-related morbidity and mortality. Natural products play a crucial role in the prevention and treatment of ALD. Hydroxysafflor yellow A (HSYA), a unique and primary component of Safflower (Carthamus tinctorius l.), exhibits diverse pharmacological activities. However, the impact and mechanism of HSYA on ALD have not been fully elucidated.
The purpose of this study was to employ an integrative pharmacology approach to assess the multi-targeted mechanism of HSYA against ALD.
Network pharmacology and molecular docking techniques were used to analyze the potential therapeutic signaling pathways and targets of HSYA against ALD. An ALD model in zebrafish larvae was established. Larvae were pretreated with HSYA and then exposed to ethanol. Liver injury was measured by fluorescence expression analysis in the liver-specific transgenic zebrafish line Tg (fabp10a:DsRed) and liver tissue H&E staining. Liver steatosis was determined by whole-mount oil red O staining and TG level. Additionally, an ethanol-induced hepatocyte injury model was established in vitro to observe hepatocyte damage (cell viability, ALT level), lipid accumulation (oil red O staining, TC and TG), and oxidative stress (ROS, MDA, GPx and SOD) in HepG2 cells treated with or without HSYA. Finally, qRT-PCR combined with network pharmacology and molecular docking was employed to validate the effects of HSYA on targets.
HSYA exhibited a significant, dose-dependent improvement in ethanol-induced liver injury in zebrafish larvae and HepG2 cells. Network pharmacology analysis revealed that HSYA may exert pharmacological effects against ALD through 341 potential targets. These targets are involved in various signaling pathways, including lipid metabolism and atherosclerosis, PI3K-Akt signaling pathway, MAPK signaling pathway, and ALD itself. Molecular docking studies displayed that HSYA had a strong binding affinity toward the domains of IL1B, IL6, TNF, PPARA, PPARG, HMGCR and ADH5. qRT-PCR assays demonstrated that HSYA effectively reversed the ethanol-induced aberrant gene expression of SREBF1, FASN, ACACA, CPT1A, PPARA, IL1B, IL6, TNFα, ADH5, and ALDH2 in vivo and in vitro.
This study offers a comprehensive investigation into the anti-ALD mechanisms of HSYA using an integrative pharmacology approach. The potential targets of HSYA may be implicated in enhancing ethanol catabolism, reducing lipid accumulation, mitigating oxidative stress, and inhibiting inflammatory response.
酒精性肝病（ALD）是导致全球与肝脏相关发病率和死亡率的主要原因之一。天然产物在 ALD 的预防和治疗中起着至关重要的作用。羟基红花黄色素 A（HSYA）是红花（Carthamus tinctorius l.）的独特且主要成分，具有多种药理活性。然而，HSYA 对 ALD 的影响和作用机制尚未完全阐明。
本研究采用综合药理学方法评估 HSYA 治疗 ALD 的多靶点机制。
采用网络药理学和分子对接技术分析 HSYA 治疗 ALD 的潜在治疗信号通路和靶点。建立斑马鱼幼鱼 ALD 模型。用 HSYA 预处理幼鱼，然后用乙醇暴露。通过荧光表达分析在肝脏特异性转基因斑马鱼系 Tg（fabp10a：DsRed）和肝脏组织 H&E 染色来测量肝损伤。通过全胚胎油红 O 染色和 TG 水平测定肝脂肪变性。此外，在体外建立乙醇诱导的肝细胞损伤模型，观察 HepG2 细胞中肝细胞损伤（细胞活力、ALT 水平）、脂质积累（油红 O 染色、TC 和 TG）和氧化应激（ROS、MDA、GPx 和 SOD）的变化，用或不用 HSYA 处理。最后，采用 qRT-PCR 结合网络药理学和分子对接验证 HSYA 对靶标的作用。
HSYA 显著改善了乙醇诱导的斑马鱼幼鱼和 HepG2 细胞的肝损伤，且呈剂量依赖性。网络药理学分析表明，HSYA 可能通过 341 个潜在靶点发挥抗 ALD 作用。这些靶点涉及多种信号通路，包括脂质代谢和动脉粥样硬化、PI3K-Akt 信号通路、MAPK 信号通路和 ALD 本身。分子对接研究表明 HSYA 与 IL1B、IL6、TNF、PPARA、PPARG、HMGCR 和 ADH5 的结构域具有很强的结合亲和力。qRT-PCR 检测显示，HSYA 可有效逆转乙醇诱导的 SREBF1、FASN、ACACA、CPT1A、PPARA、IL1B、IL6、TNFα、ADH5 和 ALDH2 在体内和体外的异常基因表达。
本研究采用综合药理学方法全面研究了 HSYA 的抗 ALD 机制。HSYA 的潜在靶点可能涉及增强乙醇代谢、减少脂质积累、减轻氧化应激和抑制炎症反应。
