Gou Xuan, Zhang Wei, Wang Lele, Tan Caixia, Wei Hong, Wang Xinmin, Zhang Le
College of Medicine, Shihezi University, Shihezi, China.
Xinjiang Provincial and Ethnic High Incidence Key Laboratory of Ministry of Education, Shihezi, China.
Front Pharmacol. 2025 Jul 30;16:1641068. doi: 10.3389/fphar.2025.1641068. eCollection 2025.
Sepsis-associated acute kidney injury (SA-AKI) is a frequent and severe complication in septic patients. This study combines network pharmacology with and experiments to preliminarily investigate the protective effect of chrysophanol (CHR) on SA-AKI and its mechanism, aiming to find new therapeutic targets and strategies for SA-AKI treatment.
HK-2 cells were used to investigate CHR's inhibitory effects on SA-AKI using CCK-8 assay, Hoechst33258 staining, ELISA, Western blot. experiments were performed using a septic mouse model, and the therapeutic effect of CHR on SA-AKI and its effect on macrophage polarization were investigated using Hematoxylin and Eosin staining, ELISA, Western blot, and quantitative real-time PCR. Predicting the possible differentially expressed genes and pathways of CHR protecting SA-AKI through network pharmacology. Finally, these pathways were further validated in experiments by ELISA, Western blot and indirect immunofluorescence staining.
CHR can inhibit LPS-induced injury and apoptosis in HK-2 cells, suppress the expression of inflammatory cytokines TNF-α and IL-6, and enhance its anti-apoptotic and anti-inflammatory effects on HK-2 cells through modulation of macrophages; in experiments, we obtained the same results that CHR effectively counteracted SA-AKI and played a protective role against mice exerting a protective effect. In addition, based on predictions from network pharmacology and validation from cellular experiments, CHR may exert these effects by inhibiting the NF-κB signalling pathway.
CHR may protect SA-AKI by inhibiting the NF-κB signalling pathway, promoting M2 macrophage polarisation and inhibiting M1 macrophage polarisation.
脓毒症相关急性肾损伤（SA-AKI）是脓毒症患者常见且严重的并发症。本研究将网络药理学与细胞和动物实验相结合，初步探究大黄酚（CHR）对SA-AKI的保护作用及其机制，旨在寻找SA-AKI治疗的新靶点和新策略。
采用HK-2细胞，运用CCK-8法、Hoechst33258染色、酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法研究CHR对SA-AKI的抑制作用。利用脓毒症小鼠模型进行动物实验，采用苏木精-伊红染色、ELISA、蛋白质免疫印迹法及实时荧光定量聚合酶链反应（qRT-PCR）研究CHR对SA-AKI的治疗作用及其对巨噬细胞极化的影响。通过网络药理学预测CHR保护SA-AKI可能的差异表达基因和信号通路。最后，通过ELISA、蛋白质免疫印迹法及间接免疫荧光染色在细胞实验中进一步验证这些信号通路。
CHR可抑制脂多糖（LPS）诱导的HK-2细胞损伤和凋亡，抑制炎性细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的表达，并通过调节巨噬细胞增强其对HK-2细胞的抗凋亡和抗炎作用；在动物实验中，我们得到了相同的结果，即CHR有效对抗SA-AKI，对小鼠发挥保护作用。此外，基于网络药理学预测和细胞实验验证，CHR可能通过抑制核因子κB（NF-κB）信号通路发挥这些作用。
CHR可能通过抑制NF-κB信号通路、促进M2巨噬细胞极化及抑制M1巨噬细胞极化来保护SA-AKI。
