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八味沉香散含药血清对H9c2细胞氧糖剥夺损伤的保护机制研究                                                                   Δ



          宫佰会 ,岳栋芳,李彩霞,官 敏,李永芳(青海大学医学院,西宁 810001)
                *
                                                #
          中图分类号  R972;R966      文献标志码  A      文章编号  1001-0408(2023)01-0040-07
          DOI  10.6039/j.issn.1001-0408.2023.01.08

          摘   要  目的  研究八味沉香散含药血清对H9c2细胞氧糖剥夺损伤的保护机制。方法  将H9c2细胞分为空白组、模型组和八味
          沉香散低、中、高剂量组(含药血清剂量依次为2.5、8、12 g/kg)。体外培养H9c2细胞并建立氧糖剥夺损伤模型,含药血清干预后检
          测细胞存活率,观察细胞形态,检测生化指标乳酸脱氢酶(LDH)、肌酸激酶(CK)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、呼
          吸链复合酶Ⅰ(ComplexⅠ)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)水平,检测细胞内活性氧(ROS)含量、线粒体膜电位
          和细胞凋亡情况,检测氧化应激相关蛋白[Kelch 样 ECH 关联蛋白 1(Keap1)、核转录因子 E2 相关因子 2(Nrf2)、血红素加氧酶 1
         (HO-1)、NADH氧化还原酶辅酶10(Ndufa10)、硫氧还蛋白(Trx)]、凋亡相关蛋白[B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、
          胱天蛋白酶3(Caspase-3)和细胞色素C(Cytc)]的表达。结果  与空白组比较,模型组细胞形态破损,LDH、CK和MDA水平均显著
          升高(P<0.01),CAT、ComplexⅠ、SOD、GSH-Px水平和线粒体膜电位均显著降低(P<0.01),细胞内ROS含量和凋亡率均显著升
          高(P<0.01),氧化应激相关蛋白(Keap1、Nrf2、HO-1、Ndufa10和Trx)和促凋亡相关蛋白(Bax、Caspase-3、Cytc)表达水平均显著升
          高(P<0.05),抗凋亡蛋白Bcl-2表达水平显著降低(P<0.05)。给予八味沉香散含药血清后,细胞形态改善,以上指标大部分显著
          逆转(P<0.05或P<0.01)。结论  八味沉香散含药血清对H9c2细胞氧糖剥夺损伤有较好的保护作用,其机制与降低细胞氧化损
          伤、抑制细胞凋亡相关。
          关键词  八味沉香散;心肌缺血;氧糖剥夺损伤;氧化应激;细胞凋亡


          Study on the protective mechanism of Bawei chenxiang powder containing serum on H9c2 cells injured by
          oxygen-glucose deprivation
          GONG Baihui,YUE Dongfang,LI Caixia,GUAN Min,LI Yongfang(School  of  Medicine,  Qinghai  University,
          Xining 810001, China)

          ABSTRACT    OBJECTIVE  To  study  the  protective  mechanism  of  Bawei  chenxiang  powder  containing  serum  on  H9c2  cells
          injured  by  oxygen-glucose  deprivation (OGD).  METHODS  H9c2  cells  were  divided  into  blank  group,  model  group  and  Bawei
          chenxiang powder low-dose, medium-dose and high-dose groups (the dose of drug containing serum 2.5, 8, 12 g/kg). H9c2 cells
          were  cultured  in  vitro  to  establish  OGD  model. After  intervention  with  drug-containing  serum,  survival  rate  of  cell  was  detected.
          The  cell  morphology  was  observed;  the  levels  of  lactate  dehydrogenase (LDH),  creatine  kinase (CK),  superoxide  dismutase
         (SOD),  catalase (CAT),  respiratory  chain  complex Ⅰ (Complex Ⅰ),  glutathione  peroxidase (GSH-Px)  and  malondialdehyde
         (MDA) were detected. The contents of reactive oxygen species (ROS), mitochondrial membrane potential and apoptosis were also
          detected. The expressions of oxidative stress-related proteins [Kelch ECH association protein 1 (Keap1), nuclear factor erythroid 2-
          related  factor  2 (Nrf2),  heme  oxygenase  1 (HO-1),  NADH  oxidoreductase  coenzyme  10 (Ndufa10),  thioredoxin (Trx)]  and
          apoptosis-related  proteins  [B-cell  lymphoma  2 (Bcl-2),  Bcl-2  associated  X  protein (Bax),  Caspase-3  and  cytochrome  C (Cytc)]
          were detected. RESULTS Compared with blank group, the cell morphology of model group was damaged; the levels of LDH, CK
          and  MDA  were  significantly  increased (P<0.01),  while  the  levels  of  CAT,  ComplexⅠ,  SOD  and  GSH-Px  and  mitochondrial
          membrane  potential  were  significantly  decreased (P<0.01). The  content  of  intracellular  ROS  and  apoptotic  rate  were  significantly
          increased (P<0.01);  the  expressions  of  oxidative  stress-related  proteins (Keap1,  Nrf2,  HO-1,  Ndufa10  and  Trx)  and  pro-
          apoptosis  proteins (Bax,  Caspase-3  and  Cytc)  were  significantly  increased (P<0.05),  while  the  expression  of  anti-apoptotic
                                                              protein  Bcl-2  was  significantly  decreased (P<0.05).  After
              Δ 基金项目 国家自然科学基金资助项目(No.81760761);青海省            administration  of  Bawei  chenxiang  powder  containing  serum,
          科技计划项目(No.2022-ZJ-746)
                                                              the  cell  morphology  improved,  and  most  of  the  above  indexes
             *第一作者 硕士研究生。研究方向:心血管药理学。E-mail:
          gbh1041183222@163.com                               were   significantly   reversed  (P<0.05   or   P<0.01).
              # 通信作者 教授,硕士生导师,硕士。研究方向:心血管药理学。                 CONCLUSIONS  Bawei  chenxiang  powder  containing  serum
          E-mail:345783110@qq.com                             has  a  good  protective  effect  on  H9c2  cells  damaged  by  OGD,


          · 40 ·    China Pharmacy  2023 Vol. 34  No. 1                                中国药房  2023年第34卷第1期
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