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钩藤碱固体脂质纳米粒抑制哮喘模型小鼠气道平滑肌细胞增殖

          的作用机制
                            Δ


          王 盟    1, 2* ,李 慧 ,吕传峰(1.英吉沙县人民医院医务部,新疆 喀什 844000;2.济宁市第一人民医院医务
                                    3
                            2
          部,山东 济宁 272000;3.济宁市第一人民医院药学部,山东 济宁 272000)

          中图分类号  R965      文献标志码  A      文章编号  1001-0408(2023)09-1066-05
          DOI  10.6039/j.issn.1001-0408.2023.09.08

          摘   要  目的  研究钩藤碱固体脂质纳米粒(Rhy-SLN)抑制哮喘模型小鼠气道平滑肌细胞(ASMCs)增殖的作用机制。方法  采用
          卵清蛋白+氢氧化铝过敏法制备哮喘模型小鼠,然后原代分离培养ASMCs并进行形态观察和鉴定[当ASMCs内α-平滑肌肌动蛋
          白(α-SMA)呈红色,结蛋白(Desmin)呈绿色,表明ASMCs培养成功];将细胞分为空白组(正常小鼠ASMCs)、模型组(哮喘模型小
          鼠ASMCs)、Rhy-SLN组(哮喘模型小鼠ASMCs)、细胞因子信号转导抑制蛋白1(SOCS1)过表达组(转染SOCS1过表达载体的哮
          喘模型小鼠ASMCs)、SOCS1-RNAi组(转染SOCS1-RNAi载体的哮喘模型小鼠ASMCs)、SB203580组[p38丝裂原激活的蛋白激
          酶(p38 MAPK)抑制剂,哮喘模型小鼠ASMCs]。各组加入相应含药(均为10 μmol/L)或不含药培养基培养24 h。采用MTT法检
          测ASMCs增殖,采用Western blot法检测ASMCs中α-SMA、白细胞介素1β(IL-1β)、SOCS1、p38 MAPK、磷酸化p38 MAPK(p-p38
          MAPK)蛋白表达水平。结果  小鼠原代ASMCs形态大小不一,呈不规则形、梭形、三角形;细胞内α-SMA呈红色,Desmin呈绿色,
          表明 ASMCs 培养成功。与模型组比较,Rhy-SLN 组、SOCS1 过表达组和 SB203580 组 ASMCs 吸光度值及 α-SMA、p38 MAPK
          和 p-p38 MAPK 蛋白表达水平均显著降低,SOCS1 蛋白表达水平(SB203580 组除外)显著升高(P<0.05);Rhy-SLN 组 ASMCs 中
          IL-1β蛋白表达水平显著降低(P<0.05);SOCS1-RNAi组ASMCs吸光度值及α-SMA、SOCS1、p38 MAPK和p-p38 MAPK蛋白表
          达水平均显著升高(P<0.05)。结论  Rhy-SLN可抑制AMSCs增殖,其作用机制可能与促进SOCS1过表达,抑制IL-1β、p38 MAPK
          蛋白表达有关。
          关键词  钩藤碱固体脂质纳米粒;气道平滑肌细胞;哮喘;α-平滑肌肌动蛋白;白细胞介素1β;细胞因子信号转导抑制蛋白1;p38
          丝裂原激活的蛋白激酶

          Mechanism  of  rhynchophylline  solid  lipid  nanoparticles  inhibiting  the  proliferation  of  airway  smooth
          muscle cells in asthmatic model mice
          WANG Meng ,LI Hui ,LYU Chuanfeng(1.  Dept.  of  Medical  Affairs,  Yingjisha  County  People’s  Hospital,
                      1, 2
                                2
                                                3
          Xinjiang  Kashi  844000,  China;2.  Dept.  of  Medical  Affairs,  Jining  First  People’s  Hospital,  Shandong  Jining
          272000, China;3. Dept. of Pharmacy, Jining First People’s Hospital, Shandong Jining 272000, China)
          ABSTRACT    OBJECTIVE  To  study  the  inhibitory  effect  mechanism  of  rhynchophylline  solid  lipid  nanoparticles (Rhy-SLN)  on
          the  proliferation  of  airway  smooth  muscle  cells (ASMCs)  in  asthmatic  model  mice.  METHODS  Asthma  model  was  prepared  by
          ovalbumin+calmogastrin sensitization. The primary isolation and culture of ASMCs were performed, and morphological observation
          and  identification  were  also  conducted  [when  α -smooth  muscle  actin (α -SMA)  appeared  red  and  Desmin  appeared  green  in
          ASMCs,  indicating  successful  cultivation  of ASMCs]. The  cells  were  divided  into  blank  group (ASMCs  of  normal  mice),  model
          group (ASMCs  of  asthma  model  mice),  Rhy-SLN  group (ASMCs  of  asthma  model  mice),  recombinant  suppressors  of  cytokine
          signaling 1 (SOCS1) overexpression group (ASMCs of asthma model mice transfected with SOCS1 vector), SOCS1-RNAi group
         (ASMCs of asthma model mice transfected with SOCS1-RNAi vector) and SB203580 group [p38 mitogen-activated protein kinase
         (p38 MAPK) inhibitor, ASMCs of asthma model mice]. The cells of each group were added into the corresponding culture medium
          containing  drug (10  μmol/L)  or  not  containing  drug  for  24  hours.  MTT  method  was  used  to  detect  the  proliferation  of ASMCs  in
          asthmatic  mice;  Western  blot  assay  was  used  to  detect  the  protein  expressions  of  α-SMA,  interleukin-1β (IL-1β),  SOCS1,  p38
          MAPK  and  phosphorylated  p38  MAPK (p-p38  MAPK)  in ASMCs.  RESULTS  The  primary ASMCs  of  mice  varied  in  shape  and
          size,  presenting  irregular,  spindle  and  triangular  shapes;α-SMA  appeared  red  and  Desmin  appeared  green,  indicating  successful
                                                              cultivation  of  ASMCs.  Compared  with  model  group,  ASMCs
                                                              absorbance  values  and  protein  expressions  of  α -SMA,  p38
              Δ 基金项目 新疆维吾尔自治区自然科学基金地州科学基金资助
          项目(No.2021D01F19)                                   MAPK,  and  p-p38  MAPK  were  reduced  significantly  in  Rhy-
             *第一作者 副主任药师,博士。研究方向:中医药防治哮喘。                     SLN  group,  SOCS1  overexpression  group  and  SB203580
          E-mail:wangmeng106@163.com                          group,  while  protein  expression  of  SOCS1  (except  for


          · 1066 ·    China Pharmacy  2023 Vol. 34  No. 9                              中国药房  2023年第34卷第9期
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