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载美洲大蠊提取物蛛丝蛋白膜的制备、表征及体外释药性和细胞

          毒性考察
                        Δ


                                        1, 4
          曾惠娜    1, 2, 3* ,卿 晨  1, 2, 3 ,薛楠楠 ,杨自忠  1, 2, 3 ,巫秀美  1, 2, 3 ,李和伟  1, 2, 3 ,赵 昱  1, 2, 3 # ,李启艳 (1.大理大学云
                                                                                              3, 5
          南省昆虫生物医药研发重点实验室,云南 大理 671000;2.大理大学药用特种昆虫开发国家地方联合工程研究
          中心,云南 大理 671000;3.大理大学中国西南药用昆虫及蛛形类资源开发利用协同创新中心,云南 大理
          671000;4.南京中医药大学药学院,南京 210023;5.云南省第一人民医院牙周科,昆明 650032)


          中图分类号  R943      文献标志码  A      文章编号  1001-0408(2023)02-0168-06
          DOI  10.6039/j.issn.1001-0408.2023.02.08

          摘   要  目的  制备一种载美洲大蠊提取物的蛛丝蛋白膜,并对其进行表征及体外释药性和细胞毒性考察。方法  以天然敬钊缨
          毛蛛蛛丝为原料,美洲大蠊提取物为模型药物,采用溶剂浇铸法制备载药蛛丝蛋白膜(后文称“载药膜”),同法制备不含美洲大蠊
          提取物的载药材料基质蛛丝蛋白膜(后文称“空白膜”)。经静态水接触角、傅里叶红外色谱、X-射线衍射以及扫描电镜等技术对膜
          剂进行表征,从不同角度分析膜剂结构;通过体外模拟药物在人工唾液中的释放特性评价药物的缓释性能;以MTT法验证载药膜
          的体外细胞毒性。结果  制备得到的载药膜的静态水接触角均小于90°,且小于空白膜;载药膜在1 500~1 700 cm 存在美洲大蠊
                                                                                                 -1
          提取物多肽的特征吸收峰;同时X-射线衍射和扫描电镜也证实美洲大蠊提取物已成功负载到膜剂中。该膜剂可在人工唾液中释
          药时长逾200 min。MTT实验结果显示,空白膜和载药膜作用后的细胞增殖率分别为84.6%、79.4%(均大于70%),无显著的潜在
          细胞毒性。结论  以天然蛛丝所制备的载药膜在人工唾液中具有一定的缓释效果,有望作为一种具优异生物相容性的药物缓释载
          体应用于临床。
          关键词  美洲大蠊;口腔膜剂;天然蛛丝;敬钊缨毛蛛;蛛丝蛋白膜;药物缓释;含量测定;细胞毒性

          Preparation,  characterization,  in  vitro  drug  release  property  and  cytotoxicity  of  Periplaneta  americana
          extract-loaded spider fibroin membrane
          ZENG Huina  1, 2, 3 ,QING Chen 1, 2, 3 ,XUE Nannan ,YANG Zizhong 1, 2, 3 ,WU Xiumei 1, 2, 3 ,LI Hewei 1, 2, 3 ,ZHAO
                                                     1, 4
          Yu 1, 2, 3 ,LI Qiyan (1.  Yunnan  Provincial  Key  Laboratory  of  Entomological  Biopharmaceutical  R&D,  Dali
                          3, 5
          University, Yunnan Dali 671000, China;2. National-Local Joint Engineering Research Center of Entomoceutics,
          Dali  University,  Yunnan  Dali  671000,  China;3.  Collaborative  Innovation  Center  for  the  Development  and
          Utilization of Medicinal Insect and Spider Resources in Southwest China, Dali University, Yunnan Dali 671000,
          China;4.  School  of  Pharmacy,  Nanjing  University  of  Chinese  Medicine,  Nanjing  210023,  China;5.  Dept.  of
          Periodontology, the First People’s Hospital of Yunnan Province, Kunming 650032, China)


          ABSTRACT    OBJECTIVE  To  prepare  spider  fibroin  membrane  loaded  with  Periplaneta  americana  extract,  and  investigate  its
          characterization,  in  vitro  drug  release  property  and  cytotoxicity.  METHODS  Using  natural  spider  silk  collected  from  Chilobrachys
          guangxiensis as raw material, P. americana extract as model drug, the drug-loaded spider fibroin membrane (hereinafter referred to
          as  drug-loaded  membrane)  was  prepared  by  solvent  casting  method.  The  material  matrix  spider  fibroin  membrane  without  P.
          americana  extract (hereinafter  referred  to  as  blank  membrane)  was  prepared  with  same  method.  The  membrane  structure  was
          characterized  by  static  water  contact  angle,  Fourier  infrared  chromatography,  X-ray  diffraction  and  scanning  electron  microscopy
          from  different  angles;  drug  release  characteristics  in  artificial  saliva  were  simulated  in  vitro  to  evaluate  the  drug  sustained-release
                                                              performance.  MTT  assay  was  adopted  to  validate  the
              Δ 基金项目 云南省科学技术院项目-生物资源数字化开发应用
         (No.202002AA100007);云南省“万人计划”名医专项(No.YNWR-MY-        cytotoxicity  of  drug-loaded   membrane.  RESULTS  The  drug-
          2018-017);云南省医学领军人才项目(No.L-2017019)                 loaded   membrane  was  prepared,  and  the  static  water  contact
             *第一作者 硕士研究生。研究方向:昆虫生物医药。E-mail:                  angle  was  less  than  90° ,  which  was  less  than  that  of  blank
          zhn970611@163.com
                                                              membrane.   The   drug-loaded    membrane   showed   the
              # 通信作者 教授,博士生导师,博士。研究方向:天然药物及生物
          资源研究与开发。E-mail:dryuzhao@126.com                     characteristic  absorption  peak  to  polypeptide  of  P.  americana


          · 168 ·    China Pharmacy  2023 Vol. 34  No. 2                               中国药房  2023年第34卷第2期
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