中性粒细胞凋亡在烧伤兔急性肺损伤发病中的作用

    Role of polymorphonuclear neutrophil apoptosis in rabbit acute lung injury after burns

    • 摘要: 目的:探讨中性粒细胞(PMN)凋亡在急性肺损伤(ALI)发病中的作用。方法:健康新西兰大白兔36只,随机分为对照组6只和烧伤组30只;烧伤组又随机分为5组,分别在烧伤后2、4、8、12、24 h 5个不同时间点处死动物,每次6只;共计6组,每组6只。观察烧伤组不同时间点兔肺组织病理,进行肺损伤评分,计算肺湿干重比(W/D),分离纯化肺泡灌洗液中的PMN,用流式细胞仪定量检测PMN凋亡和坏死细胞和呼吸爆发。结果:烧伤组各时相点均可见到肺泡腔及间质内白细胞渗出、浸润,肺泡腔内见渗出液,肺泡隔增厚,肺泡及间质充血,部分肺泡塌陷、不张,透明膜形成,且随时间延长病变加重,肺损伤的半定量评分增加(P0.01),尽管各时间点组间差异无统计学意义(P0.05),但与对照组比较差异均有统计学意义(P0.05~P0.01)。烧伤组肺W/D也随时程延长而升高(P0.01),8 h后高于对照组(P0.01);兔烧伤后2~12 h,肺灌洗液中PMN凋亡率显著降低(P0.01);PMN坏死率在烧伤后2 h显著升高(P0.05),4 h达峰值(P0.01),8 h后下降至对照组水平(P0.05);烧伤组PMN的存活率均显著高于对照组(P0.01);PMN呼吸爆发在烧伤2 h后升高,8 h到达峰值(P0.01),后下降至对照组水平。结论:ALI兔PMN从血管游出到达肺组织,在肺泡中大量聚集,且凋亡途径发生障碍,致PMN持续处于活化状态,与ALI的发病相关。

       

      Abstract: Objective:To investigate the potential role of polymorphonuclear neutrophil(PMN) apoptosis in the development of rabbit acute lung injury(ALI). Methods:Thirty-six healthy New Zealand white rabbits were randomly divided into control group(n =6) and burn group(5 time points including 2,4,8,12,24 hours after burn, with 6 in each time point). The pathological changes of lung tissue were observed. Lung injury scores(LIS) and wet-dry weight ratio(W/D) of rabbit lung tissue were calculated. PMNs were isolated and purified in bronchoalveolar lavage fluid. PMN apoptosis, necrosis, survival cell ratio and the changes of respiratory burst function were measured by flow cytometry. Results:There were significant changes in pathological section of rabbit lung tissue in burn groups at different time. It was observed that alveolar spaces and interstitial edema were occurred with massive infiltration of PMN, pulmonary architecture was damaged at 2 hours, and worsened with prolongation of time. LIS and W/D were increased at 2 hours and reached the peak at 24 hours, the differences were significant after 8 hours between control group and burn group(P 0. 01). PMN apoptosis in rabbit lung lavage fluid in burn group were decreased(P 0. 01),PMN necrosis rate was significantly increased at 2 hours(P0. 05), reached peak at 4 hours(P0. 01),restored to normal level at 8 hours(P0.05). PMN survival rate in the 5 burn groups was higher than in control group(P 0.01). The Respiratory burst of PMNs was significantly increased at 2 hours(P 0. 05),and the peak value was appeared at 8 hours(P0. 01). Conclusions:PMN congregates in alveolar spaces from pulmonary blood vessel during the pathogenesis of ALI, and apoptotic pathway is disturbed, which induce persistent activation of PMN. It may play an important role in the pathogenesis of ALI.

       

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