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柿叶提取物对H O 诱导IEC-6细胞铁死亡的影响及机制研究                                                               Δ
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          张雪侠 ,周 敏 ,周红艳 ,王莉飞 ,李华妮 ,刘长河 ,徐洪德 ,张明利 (1. 河南省中西医结合医院中药
          研究所,郑州 450004;2.郑州大学药学院,郑州 450001;3.河南中西医结合医院检验科,郑州 450004;
          4.郑州大学第二附属医院肾病科,郑州 450014;5.河南省中西医结合医院肺病科,郑州 450004)
          中图分类号  R965      文献标志码  A      文章编号  1001-0408(2025)01-0064-07
          DOI  10.6039/j.issn.1001-0408.2025.01.11
          摘   要  目的  探讨柿叶(PL)提取物对过氧化氢(H2O2 )诱导IEC-6细胞铁死亡的影响及潜在机制。方法  以IEC-6细胞为对象,考
          察铁死亡抑制剂 ferrostatin-1 对 H2O2诱导 IEC-6 细胞活力的影响;将 IEC-6 细胞分为对照组、H2O2组、H2O2+PL 25 μg/mL 组、H2O2+
          PL 50 μg/mL 组,检测其氧化应激指标[丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和活性氧(ROS)水平]、线粒体膜电位
         (MMP)以及核转录因子红系 2 相关因子 2(Nrf2)、血红素氧合酶 1(HO-1)、还原型烟酰胺腺嘌呤二核苷酸磷酸/醌氧化还原酶 1
         (NQO-1)、胱氨酸/谷氨酸反向转运体(xCT)、谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链(FTH)mRNA及蛋白的表达情况。结果
          铁死亡抑制剂 ferrostatin-1 可显著升高 H2O2诱导细胞的存活率(P<0.01)。与对照组比较,H2O2组细胞 MDA 含量和 ROS 水平,
          Nrf2、NQO-1 mRNA及Nrf2、HO-1蛋白的表达均显著升高或上调;SOD活性和MMP,xCT、GPX4、FTH mRNA及GPX4、FTH蛋白
          的表达均显著降低或下调(P<0.01或P<0.05)。与H2O2组比较,H2O2+PL 25、50 μg/mL组细胞的氧化应激指标均有不同程度回
          调,MMP均显著升高,Nrf2、HO-1、NQO-1,xCT、GPX4、FTH mRNA及蛋白的表达均有不同程度上调,部分指标组间比较差异有统
          计学意义(P<0.01 或 P<0.05)。结论  PL 提取物能减轻 H2O2所导致的细胞线粒体膜损伤、ROS 异常堆积,上述作用可能与激活
          Nrf2/HO-1信号通路从而抑制铁死亡有关。
          关键词  柿叶提取物;溃疡性结肠炎;铁死亡;Nrf2/HO-1信号通路


          Effects and mechanism of persimmon leaf extract on IEC-6 cell ferroptosis induced by H2O2
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          ZHANG Xuexia ,ZHOU Min ,ZHOU Hongyan ,WANG Lifei ,LI Huani ,LIU Changhe ,XU Hongde ,
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          ZHANG Mingli(1.  Institute  of  Chinese  Medicine,  Henan  Integrative  Medicine  Hospital,  Zhengzhou  450004,
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          China;2.  School  of  Pharmacy,  Zhengzhou  University,  Zhengzhou  450001,  China;3.  Dept.  of  Clinical
          Laboratory, Henan Integrative Medicine Hospital, Zhengzhou 450004, China;4. Dept. of Nephropathy, Second
          Affiliated  Hospital  of  Zhengzhou  University,  Zhengzhou  450014,  China;5.  Dept.  of  Pulmonary,  Henan
          Integrative Medicine Hospital, Zhengzhou 450004, China)
          ABSTRACT    OBJECTIVE To investigate the effects and potential mechanism of persimmon leaf (PL) extract against ferroptosis
          induced by H2O2 in IEC-6 cells. METHODS Using IEC-6 cells as object, the effects of ferroptosis inhibitor ferrostatin-1 on IEC-6
          cell  viability  induced  by  H2O2 were  investigated;  IEC-6  cells  were  divided  into  control  group,  H2O2  group,  H2O2+PL  25  μg/mL
          group  and  H2O2+PL  50  μg/mL  group.  The  levels  of  oxidant  stress  indexes  [content  of  malondialdehyde (MDA),  activity  of
          superoxide dismutase (SOD), and levels of reactive oxygen species (ROS)], mitochondrial membrane potential (MMP) as well as
          mRNA and protein expressions of nuclear factor-erythroid-2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NADPH/quinone
          oxidoreductase-1 (NQO-1), cystine/glutamate anti-porter (xCT), glutathione peroxidase 4 (GPX4) and ferritin heavy chain (FTH)
          were detected. RESULTS Ferroptosis inhibitor ferrostatin-1 could significantly increase the survival rate of H2O2-induced cells (P<
          0.01).  Compared  with  the  control  group,  MDA  content,  ROS  level,  mRNA  expressions  of  Nrf2  and  NQO-1  as  well  as  protein
          expressions  of  Nrf2  and  HO-1  were  increased  or  up-regulated  significantly,  while  SOD  activity,  MMP,  mRNA  expressions  of
          xCT, GPX4 and FTH as well as protein expressions of GPX4 and FTH were decreased or down-regulated significantly (P<0.01 or
                                                              P<0.05).  Compared  with  the  H2O2  group,  oxidative  stress
              Δ 基金项目 河南省自然科学基金项目(No.242300420426);河南          indexes  of  H2O2+PL  25,  50  μg/mL  groups  were  reversed  to
          省医学科技攻关计划项目(No.LHGJ20230676);河南省中西医结合医
                                                              different  extents,  MMP  level  was  increased  significantly,  as
          院基本科研业务费立项项目(No.2404041,No.2404049)                 well as mRNA and protein expressions of Nrf2, HO-1, NQO-1,
             *第一作者 助理研究员,硕士。研究方向:中药药理及药效物质
                                                              xCT,  GPX4  and  FTH  were  up-regulated  to  different  extents;
          基础。E-mail:343499957@qq.com
              # 通信作者 主任医师,硕士。研究方向:呼吸与消化系统疾病的                  there  were  statistical  significances  in  some  indexes  between
          中医治疗。E-mail:Zhang120@126.com                        groups (P<0.01  or  P<0.05).  CONCLUSIONS  PL  extract


          · 64 ·    China Pharmacy  2025 Vol. 36  No. 1                                中国药房  2025年第36卷第1期
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