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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