血红素加氧酶-1预处理对大鼠肺泡Ⅱ型上皮细胞氧化损伤的影响

Effects of heme oxygenase-1 preconditioning on oxidative damage in alveolar epithefial type II cells in rats

目的 评价血红素加氧酶-1（HO-1）预处理对大鼠肺泡Ⅱ型上皮细胞氧化损伤的影响.方法 成年健康雄性SD大鼠,体重180～220 g,原代培养肺泡Ⅱ型上皮细胞,经鉴定后随机分为6组（n=8）：对照组（C组）,不给予任何药物,继续培养5 h;H2 O2组,加入0-5 mmol/L H2 O2,孵育3 h;不同浓度HO-1预处理组（H1-4组）,分别加入0.01、0.10、1.00、10.00μmol/L HO-1孵育2 h,然后加入0.5 mmol/L H2 O2,孵育3 h.孵育结束后,于倒置相差显微镜下观察细胞形态,并进行肺泡Ⅱ型上皮细胞计数,测定细胞活力.结果 C组、H2～4组肺泡Ⅱ型上皮细胞大部分贴壁,呈圆形,胞质均匀,胞浆内含颗粒状物质;而H2 O2组和H1组肺泡Ⅱ型上皮细胞内可见反光增强的空泡,上清液中有较多的细胞碎片.与C组比较,H2O2组和H1组细胞计数和细胞活力降低（P＜0.05）,H2～4组细胞计数和细胞活力差异无统计学意义（P＞0.05）;与H2O2组和H1组比较,H2～4组细胞计数和细胞活力升高（P＜0.05）;H2O2组和H1组间,H2～4组间细胞计数和细胞活力差异无统计学意义（P＞0.05）.结论 0.10～10.00μmol/L HO-1预处理可减轻大鼠肺泡Ⅱ型上皮细胞氧化损伤.

Objective To investigate the effects of heme oxygenase-1 （HO-1） preconditioning on oxidative damage in alveolar epithelial type Ⅱ （AE- Ⅱ） cells in rats. Methods The primarily cultured AE- Ⅱ cells isolated from male SD rats were randomly assigned to one of 6 groups （n = 8 each）： control group （group C）,H2O2 group and 4 different concentrations of HO-1 preconditioning group （group H1-4）. The cells were continuously incubated for 5 h in group C. H2O2 0.5 mmol/L was added and the cells were incubated for3 h in group H2O2.In group H1-4, HO-1 0.01, 0.10, 1.00 and 10.00 μmol/L were added and the cells were incubated for2 h, then H2O2 0.5 mmol/L was added and the cells were incubated for 3 h. After the end of incubation, the cell morphology was observed under inverted phase contrast microscope, the AE- Ⅱ cells were counted, and the cell viability was determincd. Results The most AE- Ⅱ cells were adherent and round, had homogeneous cytoplasm, and the cytoplasm contains granular materials in group C and H2-4, while the vacuoles appeared in AE- Ⅱ cells and the cell debris appearecd in the supernatant in group H2 O2 and H1 . Compared with group C, the cell count and cell viability were significantly decreased in group H2 O2 and H1 （P 〈 0.05）. Compared with group H2 O2 and H1, the cell count and cell viability were significantly increased in group H2-4 （P 〈 0.05）. There was no significant difference in the cell count and cell viability between group H2O2 and H1, and among group H2～4（P 〉 0.05） .Conclusion Preconditioning with 0.10-10.00 μmol/L HO-1 can reduce oxidative damage in rat AE- Ⅱ cells.