高氧对早产鼠肺泡Ⅱ型上皮细胞的影响及神经肽P物质的保护作用.docVIP

高氧对早产鼠肺泡Ⅱ型上皮细胞的影响及神经肽P物质的保护作用 黄波，付红敏，杨鸣，方芳 ，匡凤梧，许峰 (重庆医科大学附属儿童医院急诊科，重庆 400014) 摘要： 目的 探讨高氧对离体培养的早产大鼠Ⅱ型肺泡上皮细胞(type II alveolar epithelial cells，AECⅡ)的影响及感觉神经肽P物质(substance P，SP)的保护作用。 方法 （1）分离纯化原代早产鼠AECⅡ，随机分为:空气暴露组、高氧暴露组、SP干预空气暴露组、SP干预高氧暴露组，空气暴露组氧体积分数为0.21（21%），高氧暴露组氧体积分数为0.95(95%)，SP干预组于暴露前加入SP 1′10-6mol/L,在置于氧体积分数为0.21(21%)和0.95(95%)中各组分别暴露12、24、和48h，电镜观察AECⅡ的形态变化；XTT法及流式细胞仪测定其增殖率和凋亡率。(2)剖宫取出SD大鼠孕2l d(足月为22 d)早产鼠，随机分为空气暴露组和高氧暴露组，空气暴露组氧体积分数为0.21(21%)，高氧暴露组氧体积分数为0.95(95%)，分别于暴露3，7，14 d后，取肺组织测定其SP含量。结果 与空气暴露组比较，高氧组暴露12、24、48h 后AECⅡ增殖率明显降低，凋亡率明显增加，而SP干预后其增殖率明显增加，凋亡率明显下降，形态学的损伤也有明显的改善。高氧暴露3、7、14 d 与空气暴露组比较肺组织SP含量显著降低，且随高氧暴露的延长,SP含量降低明显。结论 P物质可促进AECⅡ的增殖并抑制AECⅡ的凋亡，在高氧暴露下对AECⅡ可起到保护作用，高氧暴露可导致肺组织SP含量的变化。 关键词：P物质;高氧;Ⅱ型肺泡上皮细胞 Effect of hyperxia on the type II alveolar epithelial cells of prematural rats and protection of neuropeptide substance P HUANG Bo , FU Hong ming ， Yang Ming, FANG Fang, KUANG Feng-wu, , XU Feng, PICU, Children,s Hospital , Chongqing Medical University, Chongqing Corresponding author：XU Feng Abstract： Objective To investigate the effect of hyperxia on prematural rats type II alveolar epithelial cells (AECⅡ)cultured in vitro and the protection of neuropeptide substance P (SP). Methods I. Prematural rats AECⅡ was isolated and purified and randomly divided into following groups, including air exposure group, hyperxia exposure group and SP treatment group.Oxygen capacity fraction was 21% in air group compared to 95% in hyperxia group.SP treatment group was added 1×10-6mol/L SP before exposed to 21% and 95% oxygen for 12,24 and 48h respectively. Morphologic change of AECⅡ was observed under electron microscope.XTT and flow cytometry were employed to detect growth rate and apoptosis of AECⅡ .II. Prematural SD rats(full term is 22 d) which were 21d old were taken out by opening wombs.They were divided randomly into two groups including air exposure group(21% oxygen), hyperxia exposure group (95% oxygen).The two groups were exposed to different o