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·药学研究·


          艳山姜总黄酮改善无水乙醇致胃黏膜细胞损伤的作用及机制
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          魏 晴 ,薛 娟,梁珊珊(贵州中医药大学药学院,贵阳 550025)
          中图分类号  R965      文献标志码  A      文章编号  1001-0408(2023)22-2701-07
          DOI  10.6039/j.issn.1001-0408.2023.22.03

          摘  要  目的  探讨艳山姜总黄酮通过微 RNA-146a-5p(miR-146a-5p)对无水乙醇诱导的胃黏膜细胞损伤的改善作用及潜在机
          制。方法  以人胃黏膜细胞GES-1为对象,以无水乙醇诱导建立急性胃溃疡细胞模型,在考察不同质量浓度艳山姜总黄酮对细胞
          活力影响及筛选作用浓度的基础上,检测细胞中miR-146a-5p的相对表达量,细胞中肿瘤坏死因子受体关联因子6(TRAF6)、核因

          子 κB p65(NF-κB p65)、肿瘤坏死因子 α(TNF-α)蛋白的表达水平,以及细胞上清液中白细胞介素 1β(IL-1β)、IL-6、前列腺素 E2
         (PEG2 )水平;验证miR-146a-5p与TRAF6的靶向关系,并观察过表达miR-146a-5p、敲减TRAF6对模型细胞上清液中IL-1β、IL-6、
          PEG2水平的影响,以及敲减miR-146a-5p对艳山姜总黄酮抗胃溃疡作用的影响。结果  与空白组比较,模型组细胞中miR-146a-5p
          的相对表达量和上清液中PGE2水平均显著降低(P<0.01),细胞中TRAF6、NF-κB p65、TNF-α蛋白的表达水平和上清液中IL-1β、
          IL-6水平均显著升高(P<0.01)。与模型组比较,模型+艳山姜总黄酮(60 mg/L)组细胞中miR-146a-5p的相对表达量和上清液中
          PGE2水平均显著升高(P<0.01),细胞中TRAF6、NF-κB p65、TNF-α蛋白的表达水平和上清液中IL-1β、IL-6水平均显著降低(P<
          0.05或P<0.01)。miR-146a-5p与TRAF6存在靶向关系且呈负相关;过表达miR-146a-5p或敲减TRAF6后,细胞上清液中IL-1β、
          IL-6水平均显著降低,PGE2水平显著升高(P<0.05)。敲减miR-146a-5p后,艳山姜总黄酮作用下的细胞上清液中IL-1β、IL-6水平
          和细胞中TRAF6蛋白的表达水平均显著升高,PGE2水平显著降低(P<0.05)。结论  艳山姜总黄酮对无水乙醇致胃黏膜细胞损伤
          有一定的改善作用,其机制可能与上调miR-146a-5p的表达、抑制TRAF6的表达,进而抑制相关炎症因子的分泌有关。
          关键词  艳山姜总黄酮;胃黏膜细胞;微RNA-146a-5p;肿瘤坏死因子受体关联因子6;抗胃溃疡作用

          Improvement  effect  and  mechanism  of  total  flavonoids  from  Alpinia  zerumbet  on  gastric  mucosa  injury
          induced by absolute ethanol
          WEI Qing,XUE Juan,LIANG Shanshan(College  of  Pharmacy,  Guizhou  University  of  Traditional  Chinese
          Medicine, Guiyang 550025, China)

          ABSTRACT   OBJECTIVE To explore the improvement effect and potential mechanism of total flavonoids from Alpinia zerumbet
          on gastric mucosa injury induced by absolute ethanol through microRNA-146a-5p (miR-146a-5p). METHODS Using human gastric
          mucosa GES-1 cells as objects, the acute gastric ulcer model was established by absolute ethanol; based on the investigation of the
          effects of different concentrations of total flavonoids from A. zerumbet on cell activity and the selection of action concentration, the
          relative  expression  level  of  miR-146a-5p  in  GES-1  cells  was  detected,  the  protein  expressions  of  tumor  necrosis  factor (TNF)
          receptor-associated factor 6(TRAF6), nuclear factor-κB p65 (NF-κB p65) and TNF-α were detected, and the levels of interleukin-
          1β (IL-1β),  IL-6  and  prostaglandin  E2 (PGE2 )  in  cell  supernatant  were  determined. The  targeting  relationship  between  miR-146a-
          5p  and  TRAF6  was  verified;  the  effects  of  overexpressed  miR-146a-5p  and  TRAF6  knockdown  on  the  levels  of  IL-1β,  IL-6  and
          PEG2 in supernatant of model cells as well as the effects of miR-146a-5p knockdown on anti-gastric ulcer effect of total flavonoids
          from A.  zerumbet  were  observed.  RESULTS  Compared  with  the  blank  group,  the  relative  expression  of  miR-146a-5p  in  cells  and
          the  level  of  PGE2  in  cell  supernatant  were  decreased  significantly  in  the  model  group (P<0.01),  while  the  protein  expressions  of
          TRAF6,  NF-κB  p65  and  TNF-α  in  cells  and  the  levels  of  IL-1β  and  IL-6  in  cell  supernatant  were  increased  significantly (P<
          0.01).  Compared  with  the  model  group,  the  relative  expression  of  miR-146a-5p  in  cells  and  the  level  of  PGE2  in  cell  supernatant
                                                             were  increased  significantly  in  model+A.  zerumbet  total
             Δ 基金项目 贵州省科技计划项目(No. 黔科合基础-ZK〔2022〕一            flavonoids (60  mg/L)  group (P<0.01),  while  the  protein
          般 492;No. 黔科合基础〔2020〕1Y389);国家自然科学基金项目(No.         expressions  of  TRAF6,  NF- κB  p65  and  TNF- α  in  cells  and
          82260767)                                          the levels of IL-1β and IL-6 in cell supernatant were decreased
             *第一作者 讲师,博士。研究方向:中药药效物质基础及作用机
                                                             significantly (P<0.05  or  P<0.01).  There  was  a  targeted
          制。E-mail:997845460@qq.com
             # 通信作者 讲师,博士。研究方向:中药材加工及中药炮制机制。                 relationship  and  a  negative  correlation  between  miR-146a-5p
          E-mail:3113836821@qq.com                           and  TRAF6.  After  overexpression  of  miR-146a-5p  or  TRAF6


          中国药房  2023年第34卷第22期                                              China Pharmacy  2023 Vol. 34  No. 22    · 2701 ·
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