Zhang Junli, Hao Liyuan, Li Shenghao, He Ying, Zhang Yang, Li Na, Hu Xiaoyu
Chengdu University of Traditional Chinese Medicine, No. 37 Shi-er-qiao Road, Chengdu 610075, Sichuan Province, PR China; Department of Infectious Diseases, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, 155 Hanzhong Road, Qinghuai District, Nanjing, Jiangsu 210029, PR China.
Chengdu University of Traditional Chinese Medicine, No. 37 Shi-er-qiao Road, Chengdu 610075, Sichuan Province, PR China; Department of Infectious Diseases, Hospital of Chengdu University of Traditional Chinese Medicine, No. 39 Shi-er-qiao Road, Chengdu 610072, Sichuan Province, PR China.
Phytomedicine. 2025 Feb;137:156374. doi: 10.1016/j.phymed.2025.156374. Epub 2025 Jan 5.
Acute liver failure (ALF) has a high mortality rate, and despite treatment advancements, long-term outcomes remain poor.
This study explores the therapeutic targets and pathways of Sini Decoction plus Ginseng Soup (SNRS) in ALF using bioinformatics and network pharmacology, focusing on its impact on macrophage polarization through glucose metabolism reprogramming. The efficacy of SNRS was validated in an LPS/D-GalN-induced ALF model, and its optimal concentration was determined for in vitro macrophage intervention.
Differentially expressed genes (DEGs) in HBV-induced and acetaminophen-induced ALF were identified from GEO datasets. The correlation between target gene expression and immune cell infiltration in ALF liver tissue was analyzed. AST, ALT, TNF-α, HMGB1, IL-1β, IL-6, and IL-10 levels were measured, and liver histopathology was assessed. Macrophage polarization was analyzed via immunofluorescence, flow cytometry, and Western blot. Glycolysis-related enzymes and metabolites, including HK2, PFK-1, PKM2, and LDHA, were quantified. Cellular ultrastructure was examined by transmission electron microscopy.
Five key glycolysis-regulating genes (HK2, CDK1, SOD1, VEGFA, GOT1) were identified, with significant involvement in the HIF-1 signaling pathway. Immune infiltration was markedly higher in ALF liver tissue. SNRS improved survival, reduced ALT/AST levels, alleviated liver injury, and modulated macrophage polarization by decreasing CD86 and increasing CD163 expression. In vitro, SNRS inhibited LPS-induced inflammatory cytokine release, lactate production, p-mTOR/mTOR ratio, and HIF-1α expression.
SNRS modulates macrophage polarization and glucose metabolism reprogramming via the mTOR/HIF-1α pathway, showing promise as a treatment for ALF.
急性肝衰竭（ALF）死亡率高，尽管治疗有所进展，但长期预后仍然不佳。
本研究运用生物信息学和网络药理学探索四逆汤加人参汤（SNRS）在ALF中的治疗靶点和途径，重点关注其通过葡萄糖代谢重编程对巨噬细胞极化的影响。在脂多糖/ D - 氨基半乳糖诱导的ALF模型中验证了SNRS的疗效，并确定了其用于体外巨噬细胞干预的最佳浓度。
从基因表达综合数据库（GEO）数据集中鉴定乙型肝炎病毒诱导和对乙酰氨基酚诱导的ALF中的差异表达基因（DEG）。分析ALF肝组织中靶基因表达与免疫细胞浸润之间的相关性。测量天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、肿瘤坏死因子-α（TNF-α）、高迁移率族蛋白B1（HMGB1）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和白细胞介素-10（IL-10）水平，并评估肝脏组织病理学。通过免疫荧光、流式细胞术和蛋白质免疫印迹分析巨噬细胞极化。对包括己糖激酶2（HK2）、磷酸果糖激酶-1（PFK-1）、丙酮酸激酶M2（PKM2）和乳酸脱氢酶A（LDHA）在内的糖酵解相关酶和代谢物进行定量。通过透射电子显微镜检查细胞超微结构。
鉴定出五个关键的糖酵解调节基因（HK2、细胞周期蛋白依赖性激酶1（CDK1）、超氧化物歧化酶1（SOD1）、血管内皮生长因子A（VEGFA）、谷草转氨酶1（GOT1）），它们显著参与缺氧诱导因子-1（HIF-1）信号通路。ALF肝组织中的免疫浸润明显更高。SNRS提高了存活率，降低了ALT/AST水平，减轻了肝损伤，并通过降低CD86表达和增加CD163表达来调节巨噬细胞极化。在体外，SNRS抑制脂多糖诱导的炎性细胞因子释放、乳酸生成、磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）/哺乳动物雷帕霉素靶蛋白（mTOR）比值以及HIF-1α表达。
SNRS通过mTOR/HIF-1α途径调节巨噬细胞极化和葡萄糖代谢重编程，有望成为治疗ALF的方法。
