目的 探讨细辛通过上调p53表达介导铁死亡致大鼠肝损伤的作用机制。方法 32只大鼠随机分为对照组和细辛低、中、高剂量(0.27、0.81、1.35 g/kg)组，每组8只，每日ig给药1次，连续28 d。末次给药2 h后采集血清样本和肝脏样本，全自动生化仪检测丙氨酸氨基转移酶(alanine aminotransferase，ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase，AST)和碱性磷酸酯酶(alkaline phosphatase，ALP)水平；酶联免疫吸附法(enzyme linked immunosorbent assay，ELISA)检测肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)和白细胞介素-1β(interleukin-1β，IL-1β)水平；比色法检测超氧化物歧化酶(superoxide dismutase，SOD)活性和还原型谷胱甘肽(reduced glutathione，GSH)、丙二醛(malonaldehyde，MDA)及Fe²⁺含量；化学荧光法检测活性氧物质(reactive oxide species，ROS)含量；苏木精-伊红染色法(hematoxylin-eosin，HE)染色观察肝脏病理变化；定量反转录-聚合酶链锁反应(quantitative reverse transcription-polymerase chain reaction，qRT-PCR)检测p53、溶质载体家族7成员11(solute carrier family 7 members 11，SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4，GPX4)、铁蛋白重链1(ferritin heavy chain 1，FTH1)、转铁蛋白受体1(transferrin receptor 1，TFR1)mRNA含量；Western blotting检测p53、SLC7A11、GPX4、FTH1、TFR1蛋白表达。结果 与对照组比较，细辛组肝功能指标(ALT、AST、ALP)和血清致炎因子(TNF-α、IL-1β)水平显著升高(P＜0.05)；肝脏出现明显炎性改变；肝组织中SOD活性和GSH水平显著降低(P＜0.05)，氧化应激水平(ROS、MDA)和Fe²⁺含量显著升高(P＜0.05)；p53和TFR1的蛋白及mRNA表达均显著上调(P＜0.05)，GPX4、SLC7A11和FTH1的蛋白及mRNA表达均显著下调(P＜0.05)。结论 细辛对大鼠肝脏有一定的损伤，其机制可能与上调p53、TFR1表达水平，下调FTH1表达水平，提高细胞内Fe²⁺含量；同时下调SLC7A11表达，使GPX4表达水平降低进而诱发肝细胞铁死亡有关。

Objective To investigate the role of Xixin (Asari Radix et Rhizoma) on hepatica damage by up-regulating p53 expression to induce ferroptosis in rats. Methods Thirty-two rats were randomly divided into blank group, Asari Radix et Rhizoma low-dose, medium-dose and high-dose groups, with eight rats in each group. The rats were given orally once a day for 28 d. Serum and liver samples were collected 2 h after the last administration, and the levels of alanine aminotransaminase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were detected by automatic biochemical analyzer. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by enzyme-linked immunosorbent assay (ELISA). The activity of superoxide dismutase (SOD) and the content of reduced glutathione (GSH), malonaldehyde (MDA) and Fe²⁺ were detected by colorimetric method. The content of reactive oxygen species (ROS) was detected by chemical fluorescence method. Hematoxylin and eosin staining (HE) was used to observe the pathological changes of liver. The mRNA levels of p53, solute carrier family 7 members 11 (SLC7A11), glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1) and transferrin receptor 1 (TFR1) were detected by qRT-PCR. The protein expressions of p53, SLC7A11, GPX4, FTH1 and TFR1 were detected by Western blotting. Results Compared with the blank group, liver function indexes (ALT, AST, ALP) and serum inflammatory factors (TNF-α, IL-1β) levels in the Asari Radix et Rhizoma groups were significantly increased (P < 0.05); the obvious inflammatory changes were observed in liver; the activity of SOD and the level of GSH in liver tissue were significantly decreased (P < 0.05); the oxidative stress levels (ROS, MDA) and Fe²⁺ content were significantly increased (P < 0.05); the mRNA and protein expressions of p53 and TFR1 were significantly up-regulated (P < 0.05), while the mRNA and protein expressions of GPX4, SLC7A11 and FTH1 were significantly down-regulated (P < 0.05). Conclusion Asari Radix et Rhizoma had toxicity to liver, the mechanism might be related to the up-regulation of p53 and TFR1 expression and the down-regulation of FTH1 level, resulting the increase of intracellular Fe²⁺ concentration. At the same time, the up-regulation of p53 expression leaded to the down-regulation of SLC7A11 and GPX4 expression, resulting the induction of ferroptosis of hepatocyte.

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甘肃省自然科学基金项目(22JR5RA585)