目的:观察利多卡因(Lido)对高氧暴露的早产大鼠肺泡Ⅱ型上皮细胞(AECⅡ)增殖和凋亡的影响,为防治早产儿高氧肺损伤提供依据。方法:原代培养的早产大鼠AECⅡ随机分为4组:空气组、高氧组、空气+Lido组、高氧+ Lido组。高氧、高氧+Lido组按5 L/min通入95%O2/5%CO2高纯混合气,10 min后密封。空气+Lido、高氧+Lido组加入20 μg/mL Lido。各组均置于培养箱(37℃,5%CO2)中培养24 h,收集各组细胞,采用流式细胞仪检测AECⅡ凋亡率和细胞周期;运用Western blot 检测增殖细胞核抗原(PCNA)蛋白表达。结果:与空气组比较,高氧组AECⅡ凋亡率增高,G2/M、S期细胞比例明显降低(P<0.01);PCNA蛋白表达明显下调(P<0.01)。而Lido干预后可使AECⅡ凋亡率降低,G2/M、S期细胞比例增高,PCNA蛋白表达上调。结论:高氧可使早产大鼠AECⅡ凋亡增加,增殖抑制; Lido对高氧所致的AECⅡ损伤有直接的抑制作用。
Abstract
OBJECTIVE: To investigate the effects of lidocaine on apoptosis and proliferation of hyperoxia-exposed type Ⅱalveolar epithelial cells (AECⅡ) from premature rats. METHODS: Primary cultured AEC Ⅱ isolated from premature rats were randomly divided into four groups: air, air+ lidocaine (20 μg/mL), 95%O2/5%CO2, and 95%O2/5%CO2+ lidocaine. The cells in each group were collected 24 hrs after culture in ordinary incubators (37℃,5%CO2). The proliferation and apoptosis of AEC Ⅱ were detected by flow cytometry. Protein levels of proliferating cell nuclear antigen (PCNA) were measured by Western blot. RESULTS: The apoptosis rate of AECⅡincreased, the percentages of G2/M and S phase cells decreased and the protein levels of PCNA decreased significantly in the group exposed to 95%O2/5%CO2 compared with the group exposed to air (P<0.01). Lidocaine treatment decreased apoptosis rate of AECⅡ, increased percentage of G2/M and S phase cells, and increased protein levels of PCNA. CONCLUSIONS: Hyperoxia can increase apoptosis and inhibit proliferation of AECⅡ in premature rats. Lidocaine may have protective effects against the AECⅡ injuries.
关键词
利多卡因 /
凋亡 /
增殖 /
肺泡Ⅱ型上皮细胞 /
高氧 /
早产大鼠
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