目的 基于核转录因子E2相关因子（nuclear factor E2-related factor 2，Nrf2）/血红素加氧酶-1（heme oxygenase-1，HO-1）/谷胱甘肽过氧化物酶4（glutathione peroxidase 4，GPX4）信号通路及血清代谢组学探讨诃子砂烫炮制前后对溃疡性结肠炎（ulcerative colitis，UC）的作用机制，揭示诃子炮制增效机制。方法采用葡聚糖硫酸钠（dextran sulfate sodium，DSS）法建立大鼠UC模型，ig给予诃子生品及砂烫品，观察大鼠一般体征、疾病活动指数（disease activity index，DAI）、结直肠长度及结肠病理状态；通过定量实时聚合酶链反应（quantitative real-time polymerase chain reaction，qRT-PCR）法检测结肠组织中肿瘤坏死因子-α（tumor necrosis factor-alpha，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、IL-10、Nrf2、HO-1、GPX4、超氧化物歧化酶（superoxide dismutase，SOD）的mRNA相对表达；通过Western blotting法检测结肠组织中Nrf2、HO-1、GPX4的蛋白相对表达；采用超高效液相色谱-串联四极杆飞行时间质谱（ultra performance liquid chromatography coupled with quadrupole time-of-fight mass spectrometry，UPLC-Q-TOF-MS）检测大鼠血清中非靶向代谢物，筛选潜在生物标志物，并结合人类代谢组学数据库（human metabolomics database，HMDB）和京都基因与基因组百科全书数据库（Kyoto encyclopedia database of genes and genomes，KEGG）分析潜在代谢通路。结果 诃子生品与砂烫品均能缓解DSS诱导的UC大鼠疾病症状、降低DAI评分、恢复结直肠长度、改善结肠组织病理损伤；与模型组比较，各组大鼠结肠中炎症因子与氧化应激因子的mRNA表达水平显著逆转（P＜0.05、0.01、0.001）；Nrf2、HO-1、GPX4的蛋白表达被回调，除生品组对GPX4的调节作用外，其余各因子均具有显著性（P＜0.05、0.01、0.001）；与诃子生品组比较，诃子砂烫品在上述作用中表现出更佳效果，其中砂烫品对 Nrf2、HO-1 mRNA表达、HO-1蛋白表达的调节作用具有显著性（P＜0.001）。共筛选得到36个潜在生物标志物，其中，诃子生品与砂烫品分别调控10、12个潜在生物标志物。诃子砂烫品通过作用于精氨酸生物合成等8条信号通路，恢复机体内代谢物的平衡。结论 诃子经砂烫炮制后，可能通过抑制炎症反应，干预Nrf2/HO-1/GPX4信号通路，调节血清代谢物与精氨酸生物合成等代谢通路，发挥更强的改善UC作用。

Objective To explore the mechanism of Terminalia chebula before and after sand stir-frying processing on ulcerative colitis (UC) based on nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/glutathione peroxidase 4 (GPX4) signaling pathway and serum metabolomics, and to reveal the enhanced efficacy mechanism of T. chebula processing. Methods UC rat model was established using the dextran sulfate sodium (DSS) method. The raw and sand stir-fried T. chebula were administered by gavage to observe the general signs, disease activity index (DAI), the length of colorectum, and colon pathological status in rats. The mRNA relative expression levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), IL-10, Nrf2, HO-1, GPX4, and superoxide dismutase (SOD) in colon tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The protein relative expression levels of Nrf2, HO-1, and GPX4 in colon tissues were detected by western blotting. Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was performed to detect the serum non-targeted metabolites in rats, screen for potential biomarkers in serum, and analyze potential metabolic pathways in combination with the human metabolome database (HMDB) and Kyoto encyclopedia of genes and genomes (KEGG). Results The raw and sand stir-fried T. chebula both mitigated the disease symptoms of DSS-induced UC rats, reduced the DAI scores, restored the length of colorectum, and improved colonic pathological damage. Compared to the model group, the mRNA expression levels of inflammatory cytokines and oxidative stress cytokines in the colon of rats were significantly reversed (P < 0.05, 0.01, 0.001). The protein expression of Nrf2, HO-1, and GPX4 was restored, with all factors except for the regulation of GPX4 by the raw T. chebula group demonstrating significant effects (P < 0.05, 0.01, 0.001). Compared to the raw T. chebula group, the sand stir-fried T. chebula demonstrated superior efficacy in the above effects. Among these effects, the sand stir-fried T. chebula significantly regulated the expression of Nrf2 and HO-1 mRNA, as well as HO-1 protein expression (P < 0.001). A total of 36 potential biomarkers were screened, with the raw and sand stir-fried T. chebula regulating 10 and 12 potential biomarkers, respectively. The sand stir-fried T. chebula affected eight signaling pathways, including arginine biosynthesis, to restore the balance of metabolites in the body.Conclusion After being processed by sand stir-frying, T. chebula may exert a stronger ameliorative effect on UC by inhibiting inflammatory responses, interfering with the Nrf2/HO-1/GPX4 signaling pathway, and regulating metabolic pathways such as serum metabolites and arginine biosynthesis.

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辽宁省教育厅高校基本科研项目（LJ212410162048）;辽宁省教育厅高校基本科研项目储备项目（2024-JYTCB-011）;辽宁中医药大学自然科学类重点项目（2021LZY034）