摘要
【摘要】目的观察高体积分数氧（高氧）暴露后肺腺癌A549细胞中小窝蛋白-1（CAV-1）和转化生长因子（TGF）-β1的动态表达，探讨CAV-1和TGF-β1表达变化在高氧肺损伤中的作用。方法复苏液氮罐中冻存的A549细胞系，体外传代培养，待其在37℃、50mL／L二氧化碳（CO：）饱和湿度培养箱中生长至接近融合时，随机分为对照组和高氧组。每组18瓶A549细胞。高氧组换液后通入3L／min的950mL／L氧气和50mL／LCO2高纯混合气，10min后密闭培养。对照组仍置于50mL／LCO2培养箱中。2组分别于12h、24h、48h后在倒置相差显微镜下观察A549细胞形态并照相。采用免疫荧光双染检测CAV-1、TGF-β1表达。结果对照组CAV-1高表达（44．25±3．23），TGF-β1低表达（10．18±2．74）；高氧组随通氧时间延长，CVA-1表达逐渐减弱（12h：35．25±1．88，24h：20．75±1．68，48h：9．86±2．80），TGF-β1表达逐渐增强（12h：17．32±1．50，24h：26．51±1．82，48h：41．94±4．47）。对照组及高氧组12h、24h、48hCAV-1的阳性表达与TGF-β1的阳性表达呈负相关（r=-0．91、-0．62，-0．53、-0．88，P均〈0．05）。结论CAV-1表达降低通过减弱对TGF-β1受体的抑制作用，激活TGF-β1信号通路，导致高氧肺损伤。
Objective To observe the expressions of Caveolin-1 （ CAV-1 ） and transforming growth factor bata 1 （TGF-β1） in human adenocarcinoma of lung A549 cells exposed to hyperoxia, and to explore the changes in CAV-1 and TGF-β1 expressions and the role in hyperoxic lung injury. Methods After the A549 human lung cancer cell lines were resuscitated,the cells were packaged and cultured in 50 mL/L CO2 culture chamber, when approaching to the condition of confluence in CO2 culture chamber（37℃,50 mL/L CO2 ） ,the cells were randomly divided into control group and hyperoxia group. Eighteen bottles of A549 cells in each group. The hyperoxia group received attacking factor that was mixed with oxygen （950 mL/L）and CO2 （50 mL/L） at a velocity of 3 L/min for 10 rain ;then the flasks were enclosed and cultured in culture chamber, while those in control group were still exposed to 5 mL/L CO2. After cells were cultured for 12 h ,24 h ,48 h ,the changes in morphology were observed under the inverted microscope, the expressions CAV-1 and TGF-β1 were detected by double immunofluorescence staining technique, followed by laser scanning under the focus microscope. Results In the control group, there was high expression of CAV-1 associated with low expression of TGF-β1 （44.25 ± 3.23,10.18 ± 2.74）. Compared with control group, the content of CAV-1 reduced continuously （ 12 h ：35.25 ± 1.88,24 h ： 20.75 ± 1.68,48 h ： 9.86 ± 2.80） under prolonged hyperoxia exposure, but TGF-β1 increased gradually （ 12 h ： 17.32 ± 1.50, 24 h ：26.51 ± 1.82,48 h ：41.94 ± 4.47 ）, and the expression of CAV-1 had highly negative relation to TGF-β1 at the same time （ r = - 0.91, - 0.62, - 0.53, - 0.88, all P 〈 0.05 ） in the control group and the hyperoxia group in 12 h,24 h,48 h. Conclusions The reduction of CAV-1 expression can attenuate the inhibitory effect of TGF-β1 and activate TGF-β1 signaling pathway,which may result in hyperoxic lung injury.
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2013年第4期308-310,共3页
Chinese Journal of Applied Clinical Pediatrics
基金
四川省教育厅科研基金（08ZA150）
