中国小儿急救医学
中國小兒急救醫學
중국소인급구의학
CHINESE PEDIATRIC EMERGENCY MEDICINE
2012年
5期
498-502
,共5页
纪卫华%聂宏光%富建华%薛辛东
紀衛華%聶宏光%富建華%薛辛東
기위화%섭굉광%부건화%설신동
高氧%肺泡Ⅱ型上皮细胞%上皮钠通道%大鼠,新生
高氧%肺泡Ⅱ型上皮細胞%上皮鈉通道%大鼠,新生
고양%폐포Ⅱ형상피세포%상피납통도%대서,신생
Hyperoxia%Alveolar epithelial type Ⅱ cells%Epithelial sodium channel%Rat,newborn
目的 探讨高氧对新生大鼠肺泡Ⅱ型上皮(alveolar epithelial typeⅡ,ATⅡ)细胞钠转运功能的影响.方法 分离提取新生大鼠ATⅡ细胞,随机分成高氧组和对照组,分别在氧浓度为85％和21％的细胞培养箱中原代培养.应用Western blot方法和全细胞膜片钳技术检测高氧暴露后上皮钠通道(epithelial sodium channel,ENaC)蛋白表达变化及高氧对阿米洛利敏感性钠电流的影响.结果 高氧使新生大鼠ATⅡ细胞中β-ENaC和γ-ENaC表达增加[β-ENaC:(1 d:0.43±0.06 vs0.32 ±0.04,P=0.047;2 d:0.73 ±0.06 vs 0.50±0.08,P=0.019;3 d:0.72±0.08 vs 0.52±0.06,P=0.027);γ-ENaC:(1 d:0.64 ±0.05 vs 0.53 ±0.05,P =0.044;2 d:0.76 ±0.03 vs 0.52 ±0.04,P =0.001;3 d:0.77±0.06 vs 0.61 ±0.05,P=0.025)],同时使阿米洛利敏感性钠电流增强(1 d:13.71±2.77 vs 8.92±1.38,P＜0.001;2 d:29.12±11.03 vs 10.41±1.80,P＜0.001),面对α-ENaC表达表现出一定的抑制作用(1 d:0.31 ±0.05 vs 0.46 ±0.05,P =0.025;2 d:0.30 ±0.01 vs 0.38 ±0.02,P =0.002;3 d:0.37 ±0.06vs 0.37 ±0.08,P=0.983).结论 高氧促进了 ATⅡ细胞ENaC钠转运功能,ENaC功能障碍并非高氧暴露后肺水肿发生的主要原因.
目的 探討高氧對新生大鼠肺泡Ⅱ型上皮(alveolar epithelial typeⅡ,ATⅡ)細胞鈉轉運功能的影響.方法 分離提取新生大鼠ATⅡ細胞,隨機分成高氧組和對照組,分彆在氧濃度為85％和21％的細胞培養箱中原代培養.應用Western blot方法和全細胞膜片鉗技術檢測高氧暴露後上皮鈉通道(epithelial sodium channel,ENaC)蛋白錶達變化及高氧對阿米洛利敏感性鈉電流的影響.結果 高氧使新生大鼠ATⅡ細胞中β-ENaC和γ-ENaC錶達增加[β-ENaC:(1 d:0.43±0.06 vs0.32 ±0.04,P=0.047;2 d:0.73 ±0.06 vs 0.50±0.08,P=0.019;3 d:0.72±0.08 vs 0.52±0.06,P=0.027);γ-ENaC:(1 d:0.64 ±0.05 vs 0.53 ±0.05,P =0.044;2 d:0.76 ±0.03 vs 0.52 ±0.04,P =0.001;3 d:0.77±0.06 vs 0.61 ±0.05,P=0.025)],同時使阿米洛利敏感性鈉電流增彊(1 d:13.71±2.77 vs 8.92±1.38,P＜0.001;2 d:29.12±11.03 vs 10.41±1.80,P＜0.001),麵對α-ENaC錶達錶現齣一定的抑製作用(1 d:0.31 ±0.05 vs 0.46 ±0.05,P =0.025;2 d:0.30 ±0.01 vs 0.38 ±0.02,P =0.002;3 d:0.37 ±0.06vs 0.37 ±0.08,P=0.983).結論 高氧促進瞭 ATⅡ細胞ENaC鈉轉運功能,ENaC功能障礙併非高氧暴露後肺水腫髮生的主要原因.
목적 탐토고양대신생대서폐포Ⅱ형상피(alveolar epithelial typeⅡ,ATⅡ)세포납전운공능적영향.방법 분리제취신생대서ATⅡ세포,수궤분성고양조화대조조,분별재양농도위85％화21％적세포배양상중원대배양.응용Western blot방법화전세포막편겸기술검측고양폭로후상피납통도(epithelial sodium channel,ENaC)단백표체변화급고양대아미락리민감성납전류적영향.결과 고양사신생대서ATⅡ세포중β-ENaC화γ-ENaC표체증가[β-ENaC:(1 d:0.43±0.06 vs0.32 ±0.04,P=0.047;2 d:0.73 ±0.06 vs 0.50±0.08,P=0.019;3 d:0.72±0.08 vs 0.52±0.06,P=0.027);γ-ENaC:(1 d:0.64 ±0.05 vs 0.53 ±0.05,P =0.044;2 d:0.76 ±0.03 vs 0.52 ±0.04,P =0.001;3 d:0.77±0.06 vs 0.61 ±0.05,P=0.025)],동시사아미락리민감성납전류증강(1 d:13.71±2.77 vs 8.92±1.38,P＜0.001;2 d:29.12±11.03 vs 10.41±1.80,P＜0.001),면대α-ENaC표체표현출일정적억제작용(1 d:0.31 ±0.05 vs 0.46 ±0.05,P =0.025;2 d:0.30 ±0.01 vs 0.38 ±0.02,P =0.002;3 d:0.37 ±0.06vs 0.37 ±0.08,P=0.983).결론 고양촉진료 ATⅡ세포ENaC납전운공능,ENaC공능장애병비고양폭로후폐수종발생적주요원인.
Objective To investigate the effect of hyperoxia on the expression and transport function of epithelial sodium channel (ENaC) in neonatal rat alveolar epithelial type Ⅱ(AT Ⅱ) cells.Methods AT Ⅱ cells were isolated from neonatal rats,and primarily cultured under hyperoxic or normoxic conditions.Western blot was applied to examine the ENaC expression,and the amiloride-sensitive Na + currents were recorded using the whole-cell patch clamp technique.Results Hyperoxia upregulate the expression of β-ENaC and γ-ENaC subunits in the neonatal rat ATⅡ cells(β-ENaC:1 d:0.43 ±0.06 vs0.32 ±0.04,P =0.047;2 d:0.73±0.06 vs 0.50±0.08,P =0.019;3 d:0.72 ±0.08 vs 0.52 ±0.06,P =0.027;γ-ENaC:1 d:0.64±0.05 vs0.53 ±0.05,P =0.044;2 d:0.76 ±0.03 vs 0.52 ±0.04,P =0.001 ;3 d:0.77 ±0.06 vs 0.61 ±0.05,P =0.025).In addition,the amiloride-sensitive Na+ currents in hyperoxia-exposed AT Ⅱ cells were also increased (1d:13.71 ±2.77 vs8.92±1.38,P＜0.001;2d:29.12±11.03 vs 10.41 ±1.80,P＜0.001),which was consistent with the upregulated expression of β-ENaC and γ-ENaC.However,the expression of α-ENaC was inhibited by hyperoxia to some extent (1 d:0.31 ± 0.05 vs 0.46 ± 0.05,P =0.025 ; 2 d:0.30 ±0.01 vs0.38±0.02,P=0.002;3d:0.37±0.06 vs 0.37 ± 0.08,P =0.983).Conclusion Hyperoxia enhanced the transport function of ENaC in neonatal rat AT Ⅱ cells.Dysfunctional transport of Na + may not be a key factor involving in pulmonary edema at the early stage of bronchopulmonary dysplasia.
