高氧对早产鼠肺泡Ⅱ型上皮细胞的影响及神经肽P .doc

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法流式细胞仪测增殖凋亡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 与组比较2、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 hyperoxia on type Ⅱ alveolar epithelial cells of prematural rats and protection of neuropeptide substance P HUANG Bo, FU Hong-min, YANG Ming, FANG Fang, KUANG Feng-wu, XU Feng (Pediatric ICU, Children’s Hospital, Chongqing Medical University, Chongqing 400014, China) Abstract: Objective To investigate the effect of hyperoxia on type Ⅱ alveolar epithelial cells (AECⅡ) isolated from prematural rats and the protection of neuropeptide substance P (SP). Methods ① Prematural rats AECⅡ were isolated, purified and randomly divided into following groups, including air exposure group, hyperoxia exposure group, SP treatment group. Oxygen capacity fraction was 21% in air group while 95% in hyperoxia group. SP treatment group was added 1×10-6 mol/L SP before AECⅡ were exposed to 21% or 95% oxygen for 12, 24 and 48 h respectively. Morphologic change of AECⅡ was observed under electron microscope. XTT and flow cytometry were employed to detect growth rate and apoptosis of AECⅡ. ② Prematural SD rats (full term is 22 d) of 21 days old were taken out by opening wombs and randomly divided into two groups: air exposure group (21% oxygen) and hyperoxia exposure group (95% oxygen). After 3, 7, 14 d of exposure, the levels of SP in lung tissues were determined. Results Comp