神经肽P物质对高氧暴露下肺泡Ⅱ型上皮细胞的影响及其对信号分子Gli1的调控  

Effect of neuropeptide substance P on the typeⅡalveolar epithelial cells exposed to hyperxia and regulation function on Gli1 signal transduction molecular

作　　者：杨林[1] 党红星[1] 刘聪[1] 方芳[1] 许峰[1] 

机构地区：[1]儿童发育疾病研究省部共建教育部重点实验室,儿科学重庆市重点实验室,重庆市"儿童发育重大疾病诊治与预防"国际科技合作基地,重庆医科大学附属儿童医院PICU,重庆400014

出　　处：《重庆医学》2012年第24期2465-2468,F0003,共5页Chongqing medicine

基　　金：国家自然科学基金资助项目(30973218)

摘　　要：目的探讨神经肽P物质(SP)对高氧暴露下早产鼠肺泡Ⅱ型上皮细胞(AECⅡ)的影响及其对SHH信号通路下游信号分子Gli1的调控作用。方法分离纯化原代早产鼠AECⅡ,随机分为空气组、高氧组、高氧+SP组。空气组和高氧组分别在21%和95%氧浓度中暴露24h,高氧+SP组于高氧暴露前加入SP 1×10-8 mol/L。电子显微镜下观察AECⅡ的形态变化,MTT法检测细胞存活率,流式细胞术检测细胞凋亡率,荧光定量PCR和Western blot法检测Gli1信号分子基因和蛋白表达水平。结果与空气组比较,高氧暴露24hAECⅡ出现明显的损伤和凋亡改变,凋亡率明显增加,细胞存活率明显降低。神经肽SP干预后可明显降低AECⅡ凋亡,细胞存活率明显升高。高氧刺激可促进SHH信号通路激活Gli1基因和蛋白表达在高氧损伤的AECⅡ表达增加,SP干预后,进一步促进Gli1的表达,其基因和蛋白表达明显增加。结论 SP是一个保护性的调控因子,可以降低高氧诱导细胞损伤和凋亡,促进细胞的存活,可能与激活SHH信号通路有关。Objective To investigate the regulatory mechanism of neuropeptide substance P（SP）in lung injury induced by hyperxia and the relationship between SP and Gli1 of SHH signal transduction pathway.Methods The primary premature rats type Ⅱ alveolar epithelial cells（AECⅡ）were isolated and purified.The cells were divided into three groups：Air group,Hyperoxia group,Hyperoxia＋SP group.SP was added in advance to reach a final concentration 1×10-8 mol/L.As following,the group was exposed to 21% or 95% oxygen for 24 h respectively.Morphologic change of AECⅡwas observed under electron microscope.MTT and Flow cytometry were employed to detect the survival and apoptosis rate.Real time RT-PCR and Western blotting were used to detect the mRNA and protein expression of Gli1 signal molecules.Results After 24 h of hyperoxia exposure,apoptosis rate of AECⅡin hyperxia group increased significantly and the survival rate decreased remarkably compared to air group.The apoptosis rate decreased obviously after the intervention of SP.Meanwhile,the morphologic injure improved significantly.Hyperxia stimulation could result in activation of SHH signal transduction pathway,and the expression of Gli1 increased remarkbly in the impaired AECⅡinduced by hyperxia.Conclusion SP could decrease the oxidative stress injury and inhibit the apoptosis of AECⅡexposed to hyperxia,has protective effect on AECⅡagainst hyperoxia,which might be associated with the activation of SHH signal transduction pathway.

关 键 词：P物质 高氧 肺泡Ⅱ型上皮细胞 GLI1 

分 类 号：R459.6[医药卫生—治疗学]