目的 基于蛋白组学探讨甘草次酸和甘草酸缓解顺铂诱导的急性肝损伤（acute liver injury，ALI）的作用及潜在分子机制。方法 50只SPF级雄性Wistar大鼠随机分为对照组、模型组、氨磷汀（200 mg/kg）组、甘草次酸（100 mg/kg）组和甘草酸（200 mg/kg）组，甘草次酸组和甘草酸组连续ig给药8 d，氨磷汀组于实验第5～8天ip给药，对照组和模型组ig生理盐水。实验第5天，除对照组外，其余大鼠单次ip顺铂溶液（8 mg/kg）诱导ALI。检测肝脏指数、肝功能及肝组织病理变化；采用Label free非标定量蛋白组学技术分析肝组织蛋白表达，筛选差异表达蛋白，进行生物信息学分析。整合网络药理学方法构建药物和疾病预测交集靶点和差异表达蛋白的蛋白质-蛋白质相互作用（protein-protein interaction，PPI）整合网络，采用cytoNCA、MCODE、cytoHubba方法筛选核心靶点，进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，采用Western blotting、免疫组化等技术验证关键靶点表达。结果 与模型组比较，甘草次酸组和甘草酸组大鼠肝脏指数和血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase，AST）活性显著降低（P＜0.05、0.01、0.001），肝组织病理损伤明显改善。蛋白组学鉴定筛选到甘草次酸和甘草酸干预顺铂诱导的ALI潜在关键差异表达蛋白各23个，富集到多个氧化还原反应、炎症和凋亡相关生物学过程。PPI整合网络显示，信号转导与转录激活因子3（signal transducer and activator of transcription 3，STAT3）、血红素氧合酶1（heme oxygenase 1，HMOX1）、蛋白激酶B1（protein kinase B1，AKT1）、B细胞淋巴瘤-2（B-cell lymphoma-2，Bcl-2）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、肿瘤蛋白p53（tumor protein p53，TP53）是甘草次酸作用网络的关键靶点，HMOX1、Bcl-2、表皮生长因子受体（epidermal growth factor receptor，EGFR）、TP53是甘草酸作用网络的关键靶点，并均可富集到磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）-Akt信号通路、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）信号通路、白细胞介素-17（interleukin-17，IL-17）信号通路、p53信号通路等。验证结果显示，甘草次酸和甘草酸均能显著上调ALI大鼠肝组织p-Akt/Akt值和Bcl-2、超氧化物歧化酶（superoxide dismutase，SOD）水平（P＜0.01、0.001），下调Akt、HMOX1、STAT3、高迁移率族蛋白B1（high mobility group box 1 protein，HMGB1）表达和TNF-α、IL-1β、丙二醛（malondialdehyde，MDA）水平（P＜0.05、0.01、0.001）。结论 甘草次酸和甘草酸均能有效缓解顺铂诱导的ALI，其保护作用可能与调控Akt/STAT3/HMOX1信号轴，进而抑制肝脏氧化损伤、细胞凋亡及炎症反应密切相关。

Objective To investigate the effects and underlying molecular mechanisms of glycyrrhetinic acid (GLG) and glycyrrhizic acid (GLA) on alleviation of cisplatin (CP)-induced acute liver injury (ALI) based on proteomics. Methods A total of 50 SPF male Wistar rats were randomly divided into control group, model group, amifostine (200 mg/kg) group, GLG (100 mg/kg) group and GLA (200 mg/kg) group. Rats in GLG group and GLA group were given intragastric administration for eight consecutive days. Rats in amifostine group was given intraperitoneal administration from the 5th to the 8th day of the experiment, control group and model group were given intragastric administration of physiological saline. On the 5th day of the experiment, except for the control group, all other rats were induced to ALI with a single dose of cisplatin solution (8 mg/kg) via ip. Liver index, liver function and pathological changes in liver tissue were detected. Label free non-standard quantitative proteomics technology was used to analyze protein expressions in liver tissue, differentially expressed proteins were screen, and bioinformatics analysis was performed. The protein-protein interaction (PPI) integration network of drug and disease prediction intersection targets and differentially expressed proteins was constructed by integrating network pharmacology methods. The core targets were screened by cytoNCA, MCODE and cytoHubba, gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed. Key target expressions were validated using Western blotting, immunohistochemistry and other techniques. Results Compared with model group, the liver index and activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of rats in GLG group and GLA group were significantly reduced (P < 0.05, 0.01, 0.001), and the pathological damage of liver tissue was significantly improved. Proteomic identification identified 23 potential key differentially expressed proteins each in cisplatin-induced ALI treated with GLG and GLA, enriched in multiple biological processes related to redox reactions, inflammation and apoptosis. PPI integrated network showed that signal transducer and activator of transcription 3 (STAT3), heme oxygenase 1 (HMOX1), protein kinase B1 (AKT1), B-cell lymphoma-2 (Bcl-2), cysteine aspartate protease-3 (Caspase-3) and tumor protein p53 (TP53) were key targets of GLG action network, HMOX1, Bcl-2, epidermal growth factor receptor (EGFR) and TP53 were key targets of GLA action network, and they can be enriched in phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, tumor necrosis factor-α (TNF-α) signaling pathway, interleukin-17 (IL-17) signaling pathway, p53 signaling pathway, etc. The validation results showed that both GLG and GLA could significantly upregulate the p-Akt/Akt value and Bcl-2, superoxide dismutase (SOD) levels in liver tissue of ALI rats (P < 0.01, 0.001), and downregulate the expressions of Akt, HMOX1, STAT3, high mobility group box 1 protein (HMGB1) and levels of TNF-α, IL-1β, malondialdehyde (MDA) in liver tissue (P < 0.05, 0.01, 0.001). Conclusion GLG and GLA could effectively alleviate cisplatin-induced ALI, and their protective effects may be closely related to regulating Akt/STAT3/HMOX1 signaling axis, thereby inhibiting liver oxidative damage, cell apoptosis and inflammatory response.

R285.5

四川省科技计划项目苗子工程项目（2021JDRC0156）；四川省大学生创新创业训练计划项目（S202413705077）；国家中医药中药制剂学实验室项目（23LHZJ02）；发育与再生四川省重点实验室开放课题（23LHZY05）