目的 探讨高脂饮食对模拟低氧环境SD大鼠肺组织内皮型一氧化氮合酶(eNOS)/一氧化氮(NO)的影响及其可能机制.方法 雄性SD大鼠60只随机分为3组:常氧组、低氧组和低氧联合高脂饮食组,经普通饮食或高脂饮食4周后,留取外周血及肺组织标本,采用全自动血细胞分析仪检测静脉血血红蛋白(Hb)浓度,TBA比色法检测血浆丙二醛(MDA)含量,WST-1法检测超氧化物歧化酶(SOD)活力,终点法和直接检测法检测血脂含量,荧光实时定量PCR检测肺组织eNOS mRNA水平,Western blot法检测肺组织eNOS 蛋白表达水平,硝酸还原酶法检测肺组织NO代谢产物硝酸盐/亚硝酸盐(NOX)水平.结果 与低氧组比较,低氧联合高脂饮食组肺组织中eNOS mRNA表达水平明显升高,蛋白表达水平无明显变化,NOX含量明显降低,血浆TCH及LDL水平明显升高;低氧联合高脂饮食组血浆SOD活力及MDA含量低于低氧组.结论 高脂饮食降低低氧环境大鼠肺组织NOX水平,提示高脂饮食可引起慢性重度低氧环境下的肺组织损伤,其机制可能与血脂异常及抗氧化应激能力不足有关.
Objective To investigate the effect of high fat diet on eNOS/NO of rat lung under hypoxia environment(both at sea level and the simulated altitude of 5 000m)and clarify the possible mechanism.Methods 60 male SD rats were randomly divided into three groups:normoxia control group,hypoxia group and combined hypoxia and high fat diet group.After 4 weeks of normal diet or high fat diet,the peripheral blood and lung specimens were collected.Automatic blood cell analyzer,WST-1 and TBA methods,end point method and direct method,real-time PCR,Western blot and Nitrale reduetase method were used to test the hemoglobin(Hb)concentration,the plasma superoxide dismutase(SOD)activity,the content of malondialdehyde(MDA),the content of blood lipid,the levels of lung eNOS mRNA and protein and the levels of lung nitrates and nitrites(NOX),respectively.Results The levels of NOX were significantly decreased in H+HFD group compared with H group(P〈0.05),and while the levels of plasma TCH and LDL and the levels of eNOS mRNA in H+HFD group were higher than those in H group.The levels of eNOS protein were no significant difference between H+HFD group and H group.The plasma SOD activity and content of MDA in H+HFD group were lower than those in the H group.Conclusion High fat diet reduce NOX levels of rat lung under hypoxia environment.These results suggest that high-fat diet may induce to impairment of lung tissue under hypoxia environment,which might be,at least in part,associated with dyslipidemia and lack of ability to resist oxidative stress.