目的 基于核轉(zhuǎn)錄因子E2相關(guān)因子（nuclear factor E2-related factor 2，Nrf2）/血紅素加氧酶-1（heme oxygenase-1，HO-1）/谷胱甘肽過氧化物酶4（glutathione peroxidase 4，GPX4）信號(hào)通路及血清代謝組學(xué)探討訶子砂燙炮制前后對(duì)潰瘍性結(jié)腸炎（ulcerative colitis，UC）的作用機(jī)制，揭示訶子炮制增效機(jī)制。方法采用葡聚糖硫酸鈉（dextran sulfate sodium，DSS）法建立大鼠UC模型，ig給予訶子生品及砂燙品，觀察大鼠一般體征、疾病活動(dòng)指數(shù)（disease activity index，DAI）、結(jié)直腸長(zhǎng)度及結(jié)腸病理狀態(tài)；通過定量實(shí)時(shí)聚合酶鏈反應(yīng)（quantitative real-time polymerase chain reaction，qRT-PCR）法檢測(cè)結(jié)腸組織中腫瘤壞死因子-α（tumor necrosis factor-alpha，TNF-α）、白細(xì)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相對(duì)表達(dá)；通過Western blotting法檢測(cè)結(jié)腸組織中Nrf2、HO-1、GPX4的蛋白相對(duì)表達(dá)；采用超高效液相色譜-串聯(lián)四極桿飛行時(shí)間質(zhì)譜（ultra performance liquid chromatography coupled with quadrupole time-of-fight mass spectrometry，UPLC-Q-TOF-MS）檢測(cè)大鼠血清中非靶向代謝物，篩選潛在生物標(biāo)志物，并結(jié)合人類代謝組學(xué)數(shù)據(jù)庫(kù)（human metabolomics database，HMDB）和京都基因與基因組百科全書數(shù)據(jù)庫(kù)（Kyoto encyclopedia database of genes and genomes，KEGG）分析潛在代謝通路。結(jié)果 訶子生品與砂燙品均能緩解DSS誘導(dǎo)的UC大鼠疾病癥狀、降低DAI評(píng)分、恢復(fù)結(jié)直腸長(zhǎng)度、改善結(jié)腸組織病理?yè)p傷；與模型組比較，各組大鼠結(jié)腸中炎癥因子與氧化應(yīng)激因子的mRNA表達(dá)水平顯著逆轉(zhuǎn)（P＜0.05、0.01、0.001）；Nrf2、HO-1、GPX4的蛋白表達(dá)被回調(diào)，除生品組對(duì)GPX4的調(diào)節(jié)作用外，其余各因子均具有顯著性（P＜0.05、0.01、0.001）；與訶子生品組比較，訶子砂燙品在上述作用中表現(xiàn)出更佳效果，其中砂燙品對(duì) Nrf2、HO-1 mRNA表達(dá)、HO-1蛋白表達(dá)的調(diào)節(jié)作用具有顯著性（P＜0.001）。共篩選得到36個(gè)潛在生物標(biāo)志物，其中，訶子生品與砂燙品分別調(diào)控10、12個(gè)潛在生物標(biāo)志物。訶子砂燙品通過作用于精氨酸生物合成等8條信號(hào)通路，恢復(fù)機(jī)體內(nèi)代謝物的平衡。結(jié)論 訶子經(jīng)砂燙炮制后，可能通過抑制炎癥反應(yīng)，干預(yù)Nrf2/HO-1/GPX4信號(hào)通路，調(diào)節(jié)血清代謝物與精氨酸生物合成等代謝通路，發(fā)揮更強(qiáng)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.

R285.5

遼寧省教育廳高?；究蒲许?xiàng)目（LJ212410162048）;遼寧省教育廳高?；究蒲许?xiàng)目?jī)?chǔ)備項(xiàng)目（2024-JYTCB-011）;遼寧中醫(yī)藥大學(xué)自然科學(xué)類重點(diǎn)項(xiàng)目（2021LZY034）