Cisplatin induced drug-induced liver injury is a common toxic side effects in tumor chemotherapy, which seriously disturbs the clinical treatment of patients. Hydrogen, as a selective antioxidant, has good adjunctive therapeutic effects in various injury related diseases, but its preventive effect and mechanism in cisplatin induced liver injury are not clear.

To study the protective effect and mechanism of hydrogen water on oxidative stress injury of hepatocytes induced by cisplatin.

From June 2022 to June 2023, human normal liver cell line WRL68 were divided into control group, cisplatin group, hydrogen water group, and hydrogen water plus cisplatin group. Control group was cultured with RPMI-1640 medium containing 10% fetal bovine serum for 48 h. Cisplatin group was cultured with RPMI-1640 medium containing 10% fetal bovine serum for 24 h, then added 3 mg/L cisplatin and continued culturing for 24 h. Hydrogen water group was cultured with 0.6 mg/L hydrogen water medium for 48 h. Hydrogen water plus cisplatin group was cultured with 0.6 mg/L hydrogen water medium for 24 h, then added 3 mg/L cisplatin and continued culturing for 24 h. Then collect cells of each gorup, and hepatocyte activity was detected with CCK-8 method. Cell apoptosis was detected by flow cytometry. Mitochondrial membrane potential (MMP) was detected via Rhodamine 123 fluorescence staining method, and intracellular reactive oxygen species (ROS) levels were detected via DCFH-DA fluorescence probe staining method, the proteins and mRNA expression of nuclear factor erythroid 2-related factor 2 (NFE2L2) and heme oxygenase-1 (HMOX1) were detected by western blotting and RT-qPCR.

Compared with the control group, the apoptosis rate and ROS level of cisplatin group increased (P<0.05), while the relative levels of MMP, NFE2L2, HMOX1 protein and mRNA decreased (P<0.05). There was no statistically significant difference in cell apoptosis rate and ROS level between hydrogen water group and control group (P>0.05), while the relative levels of MMP, NFE2L2, and HMOX1 proteins and mRNA increased (P<0.05). Compared with cisplatin group, hydrogen water plus cisplatin group decreased in cell apoptosis rate and ROS level (P<0.05), while the relative levels of MMP, NFE2L2, HMOX1 proteins and mRNA increased (P<0.05).

Hydrogen water can inhibit the oxidative stress injury in cisplatin induced hepatocytes via activating NFE2L2/HMOX1 signaling pathway, and improving MMP, then alleviate drug-induced liver injury.

顺铂诱导的药物性肝损伤是肿瘤化疗常见的毒副作用，严重影响了患者临床治疗。氢作为一种选择性抗氧化剂，在多种损伤相关的疾病中均具有良好的辅助治疗作用，但其在顺铂诱导的肝损伤中的预防作用及其机制尚未明确。

探讨氢水对顺铂致肝细胞氧化应激性损伤的保护作用及作用机制。

于2022年6月—2023年6月取人正常肝细胞株WRL68分为对照组、顺铂组、氢水组、氢水+顺铂组。对照组采用含10%胎牛血清的RPMI-1640培养基培养48 h；顺铂组采用含10%胎牛血清的RPMI-1640培养基培养24 h后，加入3 mg/L顺铂继续培养24 h；氢水组采用0.6 mg/L氢水培养基培养48 h；氢水+顺铂组采用0.6 mg/L氢水培养基培养24 h后，加入3 mg/L顺铂继续培养24 h。收集各组细胞，采用CCK-8法检测细胞活力，流式细胞仪检测细胞凋亡率，罗丹明123荧光探针染色法检测线粒体膜电位（MMP），DCFH-DA荧光探针染色法检测细胞内活性氧（ROS），Western blotting和RT-qPCR检测细胞内核因子红细胞2相关因子2（NFE2L2）、血红素加氧酶1（HMOX1）蛋白及mRNA表达水平。

与对照组相比，顺铂组WRL68细胞凋亡率、ROS水平升高（P<0.05），MMP水平及NFE2L2、HMOX1的蛋白及mRNA水平降低（P<0.05）；氢水组WRL68细胞凋亡率、ROS水平与对照组相比，差异均无统计学意义（P>0.05）；氢水组MMP水平及NFE2L2、HMOX1的蛋白及mRNA水平均高于对照组（P<0.05）。与顺铂组相比，氢水+顺铂组WRL68细胞凋亡率、ROS水平均降低（P<0.05），MMP水平及NFE2L2、HMOX1的蛋白及mRNA表达水平均升高（P<0.05）。

氢水可通过激活NFE2L2/HMOX1信号通路，提高MMP，抑制顺铂引起的肝细胞氧化应激性损伤，从而减轻药物性肝损伤。