机构地区:[1]徐州市中心医院(徐州医科大学徐州临床学院)新生儿科,江苏徐州221009
出 处:《中华实用儿科临床杂志》2021年第21期1642-1647,共6页Chinese Journal of Applied Clinical Pediatrics
基 金:江苏省青年医学人才项目(QNRC2016384);徐州市科技局科研项目(KC15SH071);徐州市中心医院博士创新团队科技项目(XZB201601)。
摘 要:目的研究多瘤促活化因子3(PEA3)在高氧诱导的Ⅱ型肺泡上皮细胞(AECⅡ)损伤中的作用及其机制。方法体外培养AECⅡ,分为高氧组和常氧组。给予高氧或空气后24 h、48 h及72 h,收取各组细胞,选取最佳造模时间为48 h。将AECⅡ分3大组:空白组、阴性对照组(转染空载)、过表达质粒组(转染PEA3),每大组均分为高氧亚组和常氧亚组。给予高氧或空气后48 h,收取各组细胞。检测活性氧(ROS)、Nod样受体蛋白3(NLRP3)、单核细胞趋化蛋白-1(MCP-1)、白细胞介素(IL)-1β、IL-6、IL-8、IL-18、表面活性物质蛋白C(SP-C)、水通道蛋白5(AQP5)、PEA3及锰超氧化物歧化酶(MnSOD)等。采用SPSS 20.0统计软件,数据比较采用t检验和重复测量方差分析。结果分组与时间的交互作用对AECⅡ内ROS、IL-1β、IL-6、IL-8、IL-18、SP-C和AQP5均有显著影响(F=19.857、20.132、23.133、18.673、28.341、27.333和34.217,均P<0.05)。在24 h、48 h和72 h,高氧组ROS分别是同时间常氧组的1.78倍、1.94倍和2.26倍(t=18.649、17.486和19.385,均P<0.05);NLRP3、MCP-1均明显增加;IL-1β、IL-6、IL-8和IL-18分别是同时间常氧组的1.33倍、1.69倍和1.65倍,1.26倍、1.56倍和2.12倍,1.13倍、1.47倍和2.34倍,1.46倍、1.72倍和1.95倍(均P<0.05);SP-C蛋白表达明显减少,AQP5蛋白表达明显增加;SP-C核酸相对含量分别比同时间常氧组减少了22%、63%和72%,差异均有统计学意义(t=3.982、16.328和20.259,均P<0.05),AQP5核酸相对含量分别是同时间常氧组的1.92倍、5.23倍和7.36倍,差异均有统计学意义(t=14.631、18.945和19.521,均P<0.05)。在24 h、48 h和72 h 3个时间点,高氧组ROS、IL-1β、IL-6、IL-8、IL-18、SP-C和AQP5比较差异均有显著差异(F=22.343、20.566、23.701、19.222、32.146、40.278和37.107,均P<0.05)。在PEA3过表达48 h,与高氧阴性对照组相比,高氧过表达质粒组AECⅡ内ROS降低了34%(t=14.635,P<0.05);NLRP3、MCP-1均减少;IL-1β、IL-6、IL-8和IL-18分别降低�Objective To investigate the role of polyomavirus enhancer activator 3(PEA3)in hyperoxia-induced injury of typeⅡalveolar epithelial cells(AECⅡ)and the underlying mechanism.Methods AECⅡcells were cultured in vitro and divided into hyperoxia group and normoxia group.After 24 h,48 h and 72 h of hyperoxia or air treatment,cells were collected and the best treatment time was selected at 48 h.AECⅡcells were divided into 3 groups:control group,negative control group(transfected with negative control)and PEA3 over expression group(transfected with PEA3 overexpression plasmid).Each group was further divided into hyperoxia subgroup and normoxia subgroup.Cells were harvested at 48 h after hyperoxia or normoxia treatment.Reactive oxygen species(ROS),Nod-like receptor domain 3(NLRP3),monocyte chemoattractant protein-1(MCP-1),interleukin(IL)-1β,IL-6,IL-8,IL-18,surfactant protein C(SP-C),aquaporins 5(AQP5),PEA3 and manganese superoxide dismutase(MnSOD)levels were detected.Differences were compared by the t-test and repeated measures analysis of variance using SPSS 20.0 statistical software.Results The interaction of grouping and treatment duration had significant effects on ROS,IL-1β,IL-6,IL-8,IL-18,SP-C and AQP5 levels in AECⅡcells(F=19.857,20.132,23.133,18.673,28.341,27.333 and 34.217,respectively,all P<0.05).At 24 h,48 h and 72 h,ROS level in hyperoxia group was 1.78,1.94 and 2.26 times higher than that in normoxia group(t=18.649,17.486 and 19.385,respectively all P<0.05).NLRP3 and MCP-1 levels were significantly upregulated in hyperoxia group.IL-1βlevel was 1.33,1.69,and 1.65 times higher in hypoxia group at 24 h,48 h and 72 h than that of normoxia group;IL-6 level was 1.26,1.56 and 2.12 timers higher;IL-8 level was 1.13,1.47 and 2.34 times higher;and IL-18 level was 1.46,1.72 and 1.95 times higher,respectively(all P<0.05).The protein expression of SP-C was downregulated,while that of AQP5 was significantly upregulated in hypoxia group.The RNA expression of SP-C was 22%,63%and 72%lower in hypoxia group than tha
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