CAJ | 학술논문

目的：探讨高氧暴露对早产新生大鼠肺组织中转录因子 NF-E2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)和胞浆蛋白伴侣分子 Keap1表达的影响。方法早产新生 SD 大鼠生后1 d 完全随机设计方法分为两组：高氧组与空气组。高氧组持续暴露于常压氧舱中,氧质量浓度＞85%；空气组置于同一室常压空气中。两组分别于高浓度氧或空气暴露后1、4、7、10、14 d 提取肺组织标本。采用石蜡包埋切片行 HE 染色,观察肺组织的病理学变化。采取 RT-PCR 方法测定 Nrf2和Keap1 mRNA 水平的动态表达。Western blot 检测 Nrf2蛋白水平的表达。结果(1)与空气组相比,高氧组4、7 d 早产鼠肺组织 Nrf2 mRNA 表达显著增强(4 d：0.314±0.064 vs.0.521±0.086,7 d：0.440±0.121 vs.0.658±0.076),10 d 出现下降趋势,14 d 明显减弱(P ＜0.05)。(2)与空气组相比,高氧组1、4 d 肺组织中 Keap1 mRNA 表达均显著增强(1 d：0.352±0.052 vs.0.547±0.075,4 d：0.363±0.074 vs.0.658±0.076)(P ＜0.05),但7 d 后呈下降趋势,10、14 d 明显减弱(P ＜0.05)。(3)与空气组相比,高氧暴露1、4 d,Nrf2蛋白水平表达稍增强(P ＞0.05)；7 d 后 Nrf2蛋白呈下降趋势,其表达较空气组减弱,但7 d 两者相比差异无统计学意义(P ＞0.05),而10、14 d 显著减弱(10 d：1.325±0.464 vs.0.755±0.348,14 d：1.662±0.474 vs.0.867±0.115)(P ＜0.05)。结论氧化暴发诱导肺组织中 Nrf2与 Keap1的异常表达,调节机体氧化应激水平,参与高氧肺损伤发病过程。高氧暴露可能通过上调 Nrf2活性,提高机体抗氧化,在减轻高氧肺损伤过程中发挥一定抗氧化作用。
목적：탐토고양폭로대조산신생대서폐조직중전록인자 NF-E2상관인자2(nuclear factor-erythroid 2-related factor 2,Nrf2)화포장단백반려분자 Keap1표체적영향。방법조산신생 SD 대서생후1 d 완전수궤설계방법분위량조：고양조여공기조。고양조지속폭로우상압양창중,양질량농도＞85%；공기조치우동일실상압공기중。량조분별우고농도양혹공기폭로후1、4、7、10、14 d 제취폐조직표본。채용석사포매절편행 HE 염색,관찰폐조직적병이학변화。채취 RT-PCR 방법측정 Nrf2화Keap1 mRNA 수평적동태표체。Western blot 검측 Nrf2단백수평적표체。결과(1)여공기조상비,고양조4、7 d 조산서폐조직 Nrf2 mRNA 표체현저증강(4 d：0.314±0.064 vs.0.521±0.086,7 d：0.440±0.121 vs.0.658±0.076),10 d 출현하강추세,14 d 명현감약(P ＜0.05)。(2)여공기조상비,고양조1、4 d 폐조직중 Keap1 mRNA 표체균현저증강(1 d：0.352±0.052 vs.0.547±0.075,4 d：0.363±0.074 vs.0.658±0.076)(P ＜0.05),단7 d 후정하강추세,10、14 d 명현감약(P ＜0.05)。(3)여공기조상비,고양폭로1、4 d,Nrf2단백수평표체초증강(P ＞0.05)；7 d 후 Nrf2단백정하강추세,기표체교공기조감약,단7 d 량자상비차이무통계학의의(P ＞0.05),이10、14 d 현저감약(10 d：1.325±0.464 vs.0.755±0.348,14 d：1.662±0.474 vs.0.867±0.115)(P ＜0.05)。결론양화폭발유도폐조직중 Nrf2여 Keap1적이상표체,조절궤체양화응격수평,삼여고양폐손상발병과정。고양폭로가능통과상조 Nrf2활성,제고궤체항양화,재감경고양폐손상과정중발휘일정항양화작용。
Objective To explore the expression of nuclear factor-erythroid 2-related factor 2(Nrf2) and the molecular chaperone of cytoplasmic Keap1 in premature newborn rats exposed to hyperoxia.Methods Completely randomized design method was performed,one-day old preterm SD rats were randomly divided into two groups:hyperoxia group and air group.The preterm SD rats in hyperoxia group were continuously exposed to oxygen(oxygen >0.85)and air group in room air.After 1 ,4,7,10,14 days of exposure,the pre-term SD rats of two groups were sacrificed,whole lung of these rats were isolated,the lung histological chan-ges were observed by HE staining.Total lung RNA was extracted,Nrf2 and Keap1 mRNA were detected by RT-PCR.Western-blot was used to detect the changes of Nrf2 protein expression.Results (1 )Compaired with air group,the expression of Nrf2 in lung tissue of hyperoxia group significantly increased after 4,7 days of exposure(4 d:0.314 ±0.064 vs.0.521 ±0.086,7 d:0.440 ±0.121 vs.0.658 ±0.076)(P <0.05 ),the general tendency decreased after 10 days,but the expression of Nrf2 became significantly weak after 14 days of exposure in hyperoxia group(P <0.05).(2)The expression of Keap1 mRNA in hyperoxia group signifi-cantly increased in 1 ,4 days(1 d:0.352 ±0.052 vs.0.547 ±0.075,4 d:0.363 ±0.074 vs.0.658 ±0.076) (P <0.05),the general tendency decreased after 7 days of exposure,but the expression of Keap1 mRNA in hyperoxia group became significantly weaker than its expression after 10,14 days of air group(P <0.05 ). (3)In comparison with air group,Nrf2 protein expression in hyperoxia group increased after 1 ,4 days of ex-posure with no significant difference(P >0.05),but had a tendency of decreasing after 7 days.On day 10, 14,its expression in hyperoxia group became significantly weak compared with that of air group(10 d:1.325 ±0.464 vs.0.755 ±0.348,14 d:1.662 ±0.474 vs.0.867 ±0.1 15 )(P <0.05 ).Conclusion Oxidation outbreak results in the abnormal expression of Nrf2 and Keap1 in the lung of premature SD rats induced by hyperoxia exposure,which adjusts the levels of oxidative stress in the body,these changes participate in the development of hyperoxia induced lung injury,the activity of Nrf2 may be increased by hyperoxia exposure, and alleviate hyperoxia lung injury in premature rats through antioxidation of Nrf2.