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目的探讨神经肽P物质(Substance P,SP)对高氧损伤早产大鼠肺泡Ⅱ型上皮细胞(Alveolar epithelial celltypeⅡ,AECⅡ)氧化/抗氧化状态的影响。方法将分离培养的原代早产大鼠AECⅡ随机分为4组:空气组(将细胞置5%CO2、21%O2的孵箱中)、高氧组(将细胞置5%CO2、95%O2的密闭氧仓中)、高氧SP组(细胞培养液中预先加入5×10-8mol/L的SP,再置高氧中)和高氧SP受体拮抗剂组(细胞培养液中预先加入5×10-8mol/L的SP和5×10-7mol/L的L703.606,再置高氧中),各组细胞培养24 h后,镜下观察细胞形态的变化;MTT法检测细胞的增殖活力;流式细胞术检测细胞周期;JC-1探针法检测细胞凋亡早期线粒体膜电位的变化;化学比色法检测细胞丙二醛(Malondialdehyde,MDA)含量、总抗氧化能力(Total antioxidative capacity,TAOC)及超氧化物歧化酶(Superoxidedismutase,SOD)的活力。结果与空气组比较,高氧组AECⅡ的增殖活力、S+G2期细胞比例、线粒体膜电位、SOD活力和TAOC均明显下降(P<0.01),MDA含量明显升高(P<0.01);与高氧组比较,高氧SP组AECⅡ的增殖活力、S+G2期细胞比例、线粒体膜电位、SOD活力和TAOC均明显升高(P<0.01),而MDA含量明显下降(P<0.01),SP受体拮抗剂可阻断该效应。结论 SP可明显减轻高氧导致的早产大鼠AECⅡ的氧化损伤,增强细胞的抗氧化能力,进而起到促进细胞增殖的作用。
Objective To investigate the effect of Substance P (SP) on the oxidation / antioxidation of Alveolar epithelial cell type Ⅱ (AEC Ⅱ) in premature rats exposed to hyperoxia. Methods AEC Ⅱ isolated from preterm premature rats was randomly divided into 4 groups: air group (5% CO2, 21% O2 in incubator), hyperoxia group (5% CO2, 95% O2 ), Hyperoxia SP group (5 × 10-8mol / L SP in cell culture medium and then high oxygen medium) and hyperoxia SP receptor antagonist group (pre-added in cell culture medium 5 × 10-8mol / L SP and 5 × 10-7mol / L L703.606, then placed in high oxygen), the cells were cultured for 24 h, the morphological changes were observed under the microscope; MTT assay Cell viability was measured by flow cytometry. Changes of mitochondrial membrane potential were detected by JC-1 probe method. Malondialdehyde (MDA) content and total antioxidant capacity antioxidative capacity, TAOC) and superoxide dismutase (SOD). Results Compared with the air group, the proliferation activity, the percentage of cells in S + G2 phase, mitochondrial membrane potential, SOD activity and TAOC in hyperoxia group were significantly decreased (P <0.01) and MDA content was significantly increased (P <0.01) Compared with hyperoxia group, the activity of AECⅡ, the percentage of cells in S + G2 phase, mitochondrial membrane potential, SOD activity and TAOC in hyperoxia group were significantly increased (P <0.01), and MDA content was significantly decreased (P <0.01) SP receptor antagonists can block this effect. Conclusion SP can significantly reduce the oxidative damage of AEC Ⅱ in premature rats induced by hyperoxia and increase the anti-oxidative capacity of cells, and then play a role in promoting cell proliferation.