摘要
目的观察FPTⅢ对哮喘大鼠气道平滑肌细胞(ASMCs)分泌IL-6及Rantes的影响。方法建立大鼠哮喘模型,采用酶消化法培养ASMCs,并经免疫组织化学、免疫荧光鉴定。取第3代ASMCs,设哮喘组、对照组,各分成5个亚组:对照空白组(不加任何干预剂)、对照PBS组(加入PBS)、对照诱导组(加入10ng/ml IL-1β、TNF-α、IFN-γ混合液)、对照干预A组(加入10μM的FPTⅢ)、对照干预B组(加入混合物刺激物24h,再加入10μM的FPTⅢ);哮喘空白组、哮喘PBS组、哮喘诱导组、哮喘干预A组、哮喘干预B组,干预方式与对照组相同。采用ELISA法检测各组ASMCs分泌IL-6及Ra nte s水平。结果α-Ac tin经荧光染色后呈绿色荧光,哮喘空白组较对照空白组荧光强度减弱。哮喘干预A、B组ASMCs分泌IL-6、Rantes水平均明显低于哮喘空白组(均P<0.05),且抑制效应呈时间依赖性。FPTⅢ浓度达5~10μM时,抑制作用达到最大。结论 FPTⅢ干预可以使哮喘大鼠ASMCs分泌IL-6及Ra nte s减少,伴随细胞内α-Ac tin的变化,进一步导致细胞表型发生变化,为哮喘治疗提供新思路。
Objective to investigate the effects of FPTⅢ on the secretion of IL-6 and Rantes in airway smooth muscle cells of asthmatic rats.Methods To establish the rat model of asthma,the enzyme digestion method is adopted to cultivate airway smooth muscle cells,identified by immunohistochemistry and immunofluorescence.After using p21Ras specific inhibitor FPTⅢ intervention,IL-6 and Rantes were determinated by ELISA.The intracellular distribution of α-Actin was observed by fluorescence microscopy.Results The fluorescence intensity of α-Actin was stained by fluorescence,and the fluorescence intensity of the blank group was weaker than that of the control group.Secretion of IL-6 and Rantes were significantly decreased （P〈0.05） in asthmatic group after using FPT Ⅲ,and the inhibitory effect was time-dependent.When the concentration of FPT Ⅲ was up to 5-10 M,the inhibitory effect reached the maximum.Conclusion FPT Ⅲ intervention can make the asthmatic rat airway smooth muscle cells secreting IL-6 and Rantes decreased,accompanied by intracellular α-Actin changes,which further leads to the change of cell phenotypes,provide new ideas for the treatment of asthma.
出处
《浙江医学》
CAS
2017年第14期1175-1178,I0003,共5页
Zhejiang Medical Journal
基金
深圳市龙岗区科技计划项目(201505143001004)
