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黄芩汤的抗须癣毛癣菌活性及作用机制研究
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          沈成英 ,罗 忠 ,章 佩 ,邓冯沂 ,申宝德 ,胡建新 [1.江西省人民医院(南昌医学院第一附属医院)药学
                 1*
          部,南昌 330006;2.南昌大学药学院,南昌 330006;3.江西省人民医院(南昌医学院第一附属医院)临床医学
          研究所,南昌 330006;4.江西中医药大学现代中药制剂教育部重点实验室,南昌 330004]
          中图分类号  R275.9;R965      文献标志码  A      文章编号  1001-0408(2024)03-0311-05
          DOI  10.6039/j.issn.1001-0408.2024.03.08


          摘  要  研究黄芩汤(HQD)的抗须癣毛癣菌活性及作用机制。方法  通过测定最小抑菌浓度(MIC)、最小杀菌浓度(MFC)、菌丝
          长度、孢子萌发率、生物量和观察菌丝超微结构评价HQD的抗须癣毛癣菌活性;通过山梨醇保护实验检测HQD对须癣毛癣菌细
          胞壁的影响;通过测定麦角固醇含量和角鲨烯环氧酶(SE)、羊毛甾醇14α-去甲基化酶(CYP51)的活性考察HQD对须癣毛癣菌细
          胞膜的影响;通过测定线粒体中苹果酸脱氢酶(MDH)、琥珀酸脱氢酶(SDH)及ATP酶(包括钠钾ATP酶、钙镁ATP酶和总ATP酶)
          的活性考察HQD对须癣毛癣菌线粒体的影响。结果  HQD对须癣毛癣菌具有显著的抑菌活性,MIC、MFC值分别为3.13、25 mg/mL。
          经HQD干预后,须癣毛癣菌菌丝长度均显著缩短(P<0.05);孢子萌发率、生物量、细胞膜中麦角固醇含量和SE、CYP51活性以及
          线粒体中 MDH、SDH、各种 ATP 酶的活性均显著降低(P<0.05);细胞结构受到了一定程度的破坏,但细胞壁的完整性没有受影
          响。结论  HQD具有显著的抗须癣毛癣菌活性,其作用机制与降低细胞膜中麦角固醇含量和SE、CYP51活性以及线粒体相关酶
          活性有关。
          关键词  黄芩汤;须癣毛癣菌;抗真菌活性;作用机制;细胞膜;线粒体

          Study on the antifungal activity and mechanism of Huangqin decoction against Trichophyton mentagrophytes
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          SHEN Chengying ,LUO Zhong ,ZHANG Pei ,DENG Fengyi ,SHEN Baode ,HU Jianxin [1.  Dept.  of
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          Pharmacy,  Jiangxi  Provincial  People’s  Hospital (the  First  Affiliated  Hospital  of  Nanchang  Medical  College),
          Nanchang  330006,  China;2.  School  of  Pharmacy,  Nanchang  University,  Nanchang  330006,  China;3.  Institute
          of  Clinical  Medicine,  Jiangxi  Provincial  People’s  Hospital (the  First  Affiliated  Hospital  of  Nanchang  Medical
          College),  Nanchang  330006,  China;4.  Key  Laboratory  of  Modern  Traditional  Chinese  Medicine  Formulations
          Co-constructed by Ministry of Education, Jiangxi University of Chinese Medicine, Nanchang 330004, China]

          ABSTRACT   OBJECTIVE  To  study  the  antifungal  activity  of  Huangqin  decoction (HQD)  against  Trichophyton  mentagrophytes
          and  explore  its  mechanism.  METHODS  Minimal  inhibitory  concentration (MIC),  minimal  fungicidal  concentration (MFC),
          mycelial length, spore germination rate, biomass and mycelium ultrastructure observation were performed to evaluate the antifungal
          activity of HQD against T. mentagrophytes. The effects of HQD on the cell wall of T. mentagrophytes were detected through sorbitol
          protection  experiment.  By  measuring  the  content  of  ergosterol  and  the  activities  of  squalene  epoxide (SE)  and  lanosterol
          14α-demethylase (CYP51), the activity of HQD on the cell membrane of T. mentagrophytes was investigated. The effects of HQD
          on  T.  mentagrophytes  mitochondria  were  investigated  by  determining  the  activities  of  malate  dehydrogenase (MDH),  succinate
          dehydrogenase (SDH),  and  ATPases (including  sodium  potassium  ATPase,  calcium  magnesium  ATPase,  and  total  ATPase).
          RESULTS  HQD  exhibited  significant  antifungal  activity  against  T.  mentagrophytes  with  MIC  of  3.13  mg/mL  and  MFC  of  25
          mg/mL.  After  intervention  with  HQD,  the  mycelial  length  of  T.  mentagrophytes  was  significantly  shortened (P<0.05);  spore
          germination  rate,  biomass,  the  content  of  ergosterol  in  the  cell  membrane,  the  activities  of  SE  and  CYP51  in  the  cell  membrane
                                                             and  MDH,  SDH  and  ATPase  in  mitochondria  were  all
             Δ 基金项目 江西省自然科学基金项目(No.20232BAB216124);          decreased  significantly (P<0.05);  cell  structure  had  been
          江西省卫生健康委科技计划项目(No.202310011);江西省中医药中青              damaged  to  a  certain  extent,  but  the  integrity  of  the  cell  wall
          年骨干人才(第四批)培养计划(No.赣中医药科教字〔2022〕7号);江西              had  not  been  affected.  CONCLUSIONS  HQD  shows
          省中医药管理局中药制剂临床安全监测和转化应用重点研究室项目                      significant  antifungal  activity  against  T.  mentagrophytes,  the
             *第一作者 主管中药师,博士。研究方向:中药新型给药系统。
                                                             mechanism  of  which  may  be  associated  with  reducing  the
          电话:0791-86891529。E-mail:984075899@qq.com
             #  通信作者 主 任 药 师 。 研 究 方 向 :医 院 药 学 。 电 话 :0791-  content of ergosterol in the cell membrane and the activities of
          86895684。E-mail:hjx0108@163.com                    SE, CYP51, and mitochondria-related enzymes.


          中国药房  2024年第35卷第3期                                                 China Pharmacy  2024 Vol. 35  No. 3    · 311 ·
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