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Mechanism of rhynchophylline solid lipid nanoparticles inhibiting the proliferation of airway smooth muscle cells in asthmatic model mice
- ZHONGGUO YAOFANG Vol. 34, Issue 9, Pages: 1066-1070(2023)
- 作者机构：
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部，山东 济宁 272000
3.济宁市第一人民医院药学部，山东 济宁 272000
- 作者简介：
- 基金信息：
DOI：10.6039/j.issn.1001-0408.2023.09.08
CLC： R965Published：15 May 2023，
Received：22 October 2022，
Revised：30 March 2023，
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王盟,李慧,吕传峰.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[J].中国药房,2023,34(09):1066-1070.
WANG Meng,LI Hui,LYU Chuanfeng.Mechanism of rhynchophylline solid lipid nanoparticles inhibiting the proliferation of airway smooth muscle cells in asthmatic model mice[J].ZHONGGUO YAOFANG,2023,34(09):1066-1070.
王盟,李慧,吕传峰.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[J].中国药房,2023,34(09):1066-1070. DOI： 10.6039/j.issn.1001-0408.2023.09.08.
WANG Meng,LI Hui,LYU Chuanfeng.Mechanism of rhynchophylline solid lipid nanoparticles inhibiting the proliferation of airway smooth muscle cells in asthmatic model mice[J].ZHONGGUO YAOFANG,2023,34(09):1066-1070. DOI： 10.6039/j.issn.1001-0408.2023.09.08.
摘要
目的
2
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
2
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
2
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
2
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。
Abstract
OBJECTIVE
2
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
2
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
2
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 MAPK， and p-p38 MAPK were reduced significantly in Rhy-SLN group， SOCS1 overexpression group and SB203580 group， while protein expression of SOCS1 （except for SB203580 group） was increased significantly （
P
＜0.05）； protein expressions of IL-1β was reduced significantly in ASMCs （
P
＜0.05）. ASMCs absorbance values and protein expressions of
α
-SMA， SOCS1， p38 MAPK and p-p38 MAPK were increased significantly in SOCS1-RNAi group （
P
＜0.05）.
CONCLUSIONS
2
Rhy-SLN can inhibit the proliferation of ASMCs， the mechanism of which may be associated with overexpression of SOCS1 and inhibiting the protein expressions of IL-1β and p38 MAPK.
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1p38丝裂原激活的蛋白激酶
Keywords
airway smooth muscle cellsasthmaα-smooth muscle actininterleukin-1βrecombinant suppressors of cytokine signaling 1p38 mitogen-activated protein kinase
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