桔梗总皂苷对小鼠气道平滑肌细胞增殖、迁移及Notch1/Jagged1通路的影响  

Effects of Platycodonis Radix Total Saponins on the Proliferation,Migration and Notch1/Jagged1 Pathway of Mouse Airway Smooth Muscle Cells

作　　者：鲍国斌 郑应升 彭峰 BAO Guobin;ZHENG Yingsheng;PENG Feng(Pediatrics Department of the First People's Hospital of Yuhang District,Hangzhou 311113,China;Zhejiang Integrated Traditional Chinese and Western Medicine Hospital,Hangzhou 310000,China)

机构地区：[1]余杭区第一人民医院儿科,杭州311113 [2]浙江省中西医结合医院,杭州310000

出　　处：《中国现代应用药学》2024年第18期2580-2587,共8页Chinese Journal of Modern Applied Pharmacy

基　　金：杭州市科技计划引导项目(20211231Y043)。

摘　　要：目的探讨桔梗总皂苷对小鼠气道平滑肌细胞(airwaysmoothmusclecells, ASMCs)增殖、迁移及Notch1/Jagged1信号通路相关蛋白表达水平的影响。方法 分离、培养小鼠原代ASMCs,按照对照组、重组小鼠血小板衍生生长因子(platelet-derived growth factor,PDGF)组,10、20、40μg·mL^(-1)桔梗总皂苷组和IL-17+40μg·mL^(-1)桔梗总皂苷组,随机分组给药。Notch1过表达慢病毒感染ASMCs,按照对照组、PDGF组、PDGF+40μg·mL^(-1)桔梗总皂苷组、PDGF+40μg·mL^(-1)桔梗总皂苷+pEGF-C1(Notch1过表达阴性对照质粒)组和PDGF+40μg·mL^(-1)桔梗总皂苷+pEGFC1/Notch1 (Notch1过表达质粒)组分组给药。CCK8、划痕试验分别检测ASMCs的增殖和迁移能力,免疫荧光检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达水平,ELISA检测ASMCs上清液中炎症相关因子水平,Western blotting检测ASMCs中Notch1/Jagged1通路相关蛋白表达水平。结果 不同剂量的桔梗总皂苷均能不同程度地抑制PDGF诱导的细胞增殖、迁移,抑制ASMCs中PCNA荧光表达增强,显著抑制上清液中IL-6、IL-8水平及Notch1、Jagged1、Hes1蛋白表达水平的上调(P<0.05或P<0.01);与40μg·mL^(-1)桔梗总皂苷组比较,IL-17+40μg·mL^(-1)桔梗总皂苷组细胞活力、迁移率及上清液中IL-6、IL-8水平均显著升高(P<0.05或P<0.01),PCNA荧光表达强度显著增强(P<0.01),Notch1、Jagged1、Hes1蛋白表达水平显著上调(P<0.05或P<0.01)。与40μg·mL^(-1)桔梗总皂苷+pEGF-C1组比较,40μg·mL^(-1)桔梗总皂苷+pEGF-C1/Notch1组ASMCs的细胞活力、迁移率及上清液中IL-6、IL-8水平均显著升高(P<0.05或P<0.01),Notch1、Jagged1、Hes1蛋白表达水平显著性上调(P<0.01)。结论 桔梗总皂苷对ASMCs的增殖和迁移有抑制作用,其作用机制可能与下调Notch1/Jagged1信号通路相关蛋白表达水平有关。OBJECTIVE To investigate the effects of Platycodonis Radix total saponins on the proliferation and migration of airway smooth muscles(ASMCs)and the expression levels of Notch1/Jagged1 signaling pathway-related proteins in asthmatic mice.METHODS Primary mouse ASMCs were isolated and cultured,and administered in randomized groups according to the control group,platelet-derived growth factor(PDGF)group,10μg·mL^(-1) Platycodonis Radix total saponins group,20μg·mL^(-1) Platycodonis Radix total saponins group,40μg·mL^(-1) Platycodonis Radix total saponins group and IL-17+40μg·mL^(-1) Platycodonis Radix total saponins group.Notch1 overexpression lentivirus infected ASMCs,according to the control group,PDGF group,PDGF+40μg·mL^(-1) Platycodonis Radix total saponins group,PDGF+40μg·mL^(-1) Platycodonis Radix total saponins+pEGF-C1(Notch1 overexpression negative control plasmid)group and PDGF+40μg·mL^(-1) Platycodonis Radix total saponins+pEGF-C1/Notch1(Notch1 overexpression plasmid)group administration.The proliferation and migration ability of ASMCs were detected by CCK8 and scratch assay,the expression level of PCAN was detected by immunofluorescence,the level of inflammation-related factors in ASMCs supernatant was detected by ELISA,and the expression level of Notch1/Jagged1 pathway-related proteins in ASMCs was detected by Western blotting.RESULTS Different doses of Platycodonis Radix total saponins can inhibit PDGF-induced cell proliferation and migration to varying degrees,inhibit the enhancement of proliferating cell nuclear antigen(PCNA)fluorescence expression in ASMCs,significantly inhibit the levels of IL-6 and IL-8 in the supernatant and the expressions of Notch1,Jagged1,and Hes1 proteins Level up(P<0.05 or P<0.01).Compared with the 40μmol·L^(-1) Platycodonis Radix total saponins group,the cell viability,mobility and the levels of IL-6 and IL-8 in the supernatant of the IL-17+40μmol·L^(-1) Platycodonis Radix total saponins group were significantly increased(P<0.05 or P<0.01),PCNA fluorescence e

关 键 词：哮喘 桔梗总皂苷 气道平滑肌细胞 增殖 迁移 Notch1/Jagged1信号通路 

分 类 号：R284.1[医药卫生—中药学]