目的 对比熟地黄水提取物（RRPAE）与生地黄水提取物（DRRAE）治疗肝缺血再灌注损伤的药效和机制，以期为临床患者更优的治疗选择提供科学依据。方法 将48只C57雄性小鼠随机分为8组（每组6只）：假手术组，模型组，RRPAE低、中、高剂量（2.5、5.0、10.0 g·kg^-1）组，DRRAE低、中、高剂量（2.5、5.0、10.0 g·kg^-1）组。ig给药，假手术组和模型组ig 0.9%氯化钠溶液，每天1次，连续1周；各组小鼠于末次给药2 h后行手术建立小鼠肝缺血再灌注损伤模型，假手术组仅开腹不缺血，再灌注后6 h取材。检测各组小鼠肝脏、脾脏系数；苏木素-伊红染色观察各组小鼠肝脏组织病理学改变；试剂盒法检测小鼠血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、乳酸脱氢酶（LDH）、一氧化氮（NO）水平；试剂盒法检测小鼠肝脏超氧化物歧化酶（SOD）、NO、丙二醛（MDA）水平；Western blotting法检测各组小鼠肝脏组织中铁死亡相关蛋白转铁蛋白（Transferrin）、溶质载体家族7成员11 （SLC7A11）、溶质载体家族39成员14 （SLC39A14）、poly结合蛋白2（PCBP2）的表达水平。培养BRL肝细胞，用50、100、150 mg·mL^-1的RRPAE处理1 h （对照组不加药）后，加野百合碱（500 μmol·L^-1）或埃拉斯汀（10 μmol·L^-1）处理24 h，试剂盒法分别检测细胞亚铁/总铁离子以及谷胱甘肽水平。结果 与模型组比较，经过RRPAE或DRRAE预给药后，给药组小鼠的肝脏、脾脏系数出现下降趋势，其中RRPAE低、高剂量组肝脏系数差异显著（P＜0.05、0.001）；病理染色结果显示模型组肝脏出现明显的缺血-充血区和肝细胞坏死，经过RRPAE与DRRAE给药后肝细胞状态均有所恢复，且前者的效果明显优于后者。生化结果显示，与模型组相比，RRPAE可显著降低血清中肝损伤指标AST、ALT、LDH水平，升高NO含量；显著降低肝脏MDA水平，升高SOD和NO水平（P＜0.05、0.01） ；而DRRAE给药后对上述肝损伤及氧化应激相关指标的恢复作用整体并不显著。Western blotting实验结果显示，与模型组比较，RRPAE能够显著抑制促进铁死亡相关蛋白Transferrin、SLC39A14的表达，上调抑制铁死亡相关蛋白SLC7A11、PCBP2的表达（P＜0.05、0.001） ，而DRRAE对铁死亡相关蛋白的调控并不明显。与模型组比较，RRPAE可上调野百合碱或铁死亡激动剂埃拉斯汀诱导的肝细胞培养基中亚铁/总铁离子以及谷胱甘肽水平（P＜0.05、0.01、0.001）。结论 熟地黄缓解肝缺血再灌注损伤效果更好，其作用机制可能与调控Transferrin、SLC7A11、SLC39A14、PCBP2蛋白的表达，抑制小鼠铁死亡进程有关。

Objective Based on comparing the efficacy and mechanism of aqueous extracts of Rehmanniae Radix Praeparata (RRPAE) and drying Rehmannia root (DRR) for the treatment of hepatic ischemia-reperfusion injury (HIRI), to provide a better therapeutic option for clinical patients. Method Totally 48 mice were randomly divided into eight groups: the sham group, the model group, the low, medium, and high dose groups of RRPAE (2.5, 5.0 and 10.0 g·kg^-1), and the low, medium, and high dose groups of DRRAE (2.5, 5.0 and 10.0 g·kg^-1), with six mice in each group. After one week of pre-administration of different doses of RRPAE or DRRAE by ig, the mouse HIRI model was established, and the HE staining was used to observe the histopathological changes in the livers of the mice in each group. The kits were used to detect the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), nitric oxide (NO) levels. The kits were used to detect mouse liver superoxide dismutase (SOD), nitric oxide (NO), and malondialdehyde (MDA) levels. Western blotting was used to detect ferroptosis-associated proteins in mouse liver tissues of each group: Transferrin, solute carrier family 7 member 11 (SLC7A11), solute carrier family 39 member 14 (SLC39A14), and poly rC binding protein 2 (PCBP2). After culturing and treating BRL hepatocytes with monocrotaline (MCT, 500 μmol·L^-1) or ferroptosis agonist erastin (10 μmol·L^-1), different doses of RRPAE (50, 100, and 150 μg·mL^-1) were administered for 24 h, and then the cellular ferrous iron/ total iron ions and glutathione content were detected. Results Compared with the model group, after pre-administration of RRPAE or DRRAE, the liver and spleen coefficients of mice in the treatment group showed a downward trend, with significant differences in liver coefficients in the low and high dose RRPAE groups (P＜0.05, 0.001). HE staining results showed significant ischemia-reperfusion region and hepatocellular necrosis in the HIRI group, and the hepatocellular status was recovered after both RRPAE and DRRAE administration, and the former was significantly better than the latter. The biochemical results showed that compared with the model group, the serum levels of AST, ALT and LDH, which were indicators of liver injury, decreased significantly, while the level of NO increased significantly; the level of hepatic MDA decreased significantly, while the levels of SOD and NO increased significantly (P＜0.05, 0.01), and the modulation effect of DRR on the above indicators of liver injury and oxidative stress was not significant. The results of Western blotting experiments showed that the RRPAE was able to significantly inhibit the expression of Transferrin and SLC39A14-related ferroptosis-promoting proteins, and down-regulate the expression of SLC7A11 and PCBP2-related ferroptosis-suppressing proteins (P＜0.05, 0.001), whereas the DRRAE did not significantly regulate the above iron-death-related proteins. At the same time, RRPAE directly inhibited the ferroptosis process of hepatocytes induced by monocrotaline or ferroptosis agonist erastin, which was accompanied by an increase in ferrous iron/total iron ions and glutathione content in the cell culture medium (P＜0.05, 0.01, and 0.001). Conclusion The medicinal type of DRR with better effect in alleviating hepatic ischemia-reperfusion injury is RRPAE, and its mechanism of action may be related to the regulation of Transferrin, SLC7A11, SLC39A14, and PCBP2 protein expression and inhibition of ferroptosis process in mice.

R285.5

国家自然科学基金面上项目（82274186）