血红素加氧酶-1对过氧化氢损伤的Ⅱ型肺泡上皮细胞凋亡的影响  

The effects of heme oxygenase-1 on apoptosis induced by hydrogen peroxide in type Ⅱ alveolar epithelium cell

作　　者：刘海霞[1] 陈淼[1] 杨秀娟[1] 戢慧[1] 陈涛[1] 陈华军[1] 

机构地区：[1]遵义医学院附属医院重症医学科,贵州563000

出　　处：《中华危重病急救医学》2014年第2期110-114,共5页Chinese Critical Care Medicine

基　　金：贵州省科技计划项目（2008-3052）;贵州省教育厅自然科学基金项目（2007-042）

摘　　要：目的探讨血红素加氧酶-1（HO-1）对过氧化氢（H202）刺激后原代培养大鼠Ⅱ型肺泡上皮细胞（AECⅡ）凋亡率、活化天冬氨酸特异性半胱氨酸蛋白酶3（easpase-3）及细胞色素C（Cyt-c）表达的影响。方法原代培养雄性sD大鼠AEC1I细胞，被随机分为4组：对照组（A组）加入生理盐水；H202损伤组（B组）加入0．5mmo]]LH，0：处理；HO一1预处理组（C组）给予0．2μmol／LHO-1预处理2h后加入0．5mmol／LH202；HO-1抑制组（D组）给予10μmol／L锌原卟啉Ⅸ（ZnppIX）预处理2h后加入0．5mmol/L H202，各组细胞处理后继续培养。于药物干预后2、6、12h采用流式细胞仪检测细胞凋亡率，用蛋白质免疫印迹试验（WesternBlot）检测活化easpase-3、Cyt-c蛋白表达。结果各组细胞凋亡率均随H202作用时间延长而逐渐升高；B组干预后各时间点AEClI凋亡率均较A组显著增加；C组AEClI凋亡率则较B组明显降低[2h：（11．46±1．47）％比（20．83±1．55）％，6h：（12．30±1．37）％比（27．14±1．53）％，12h：（12．62±1．39）％比（35．66±0．74）％，均P〈0．05]；D组2、6、12hAEC1I凋亡率[（24．33±1．36）％、（31．67±1．24）％、（36．93±2．40）％]与B组比较则差异无统计学意义。各组活化easpase-3、Cyt-C蛋白表达量均随H：0：作用时间延长而逐渐增加；B组各时间点活化easpase-3、Cyt-c蛋白表达量均较A组明显增加；c组活化easpase-3蛋白表达量（灰度值）较B组明显减少（2h：0．250±0．039比0．650±0．072，6h：0．470±0．080比0．960±0．118，12h：0．680±0．099比1．830±0．220，均P〈O．05）；Cyt-c蛋白表达量（灰度值）也较B组明显减少（2h：0．250±0．074比0．390±0．069，6h：0．340±0．043比0．670±0．120，12h：0．470±0．072比1．360±0．112，均P〈O．05）；D组活化caspase-3蛋白表达量（0．720±0．052、1．060±0．109、1．880±0．159）、Cyt-C蛋�Objective To explore the effects of heme oxygenase-1 （HO-1） on the apoptosis, expression of easpase-3 and cytochrome-C （Cyt-C） on hydrogen peroxide （H202） oxidative damage in primary cultured rat＇s type Ⅱ alveolar epithelium cell （AEC H ）. Methods AEC H from male Sprague-Dawley （SD） rats were separated and cultured. The cells were randomly divided into four groups ： control group （A group, the cells were cultured with nmTnal saline）, H202 group （B group, cultured with 0.5 mmol/L H202）, HO-1 pretreatment group （C group, pretreatment with 0.2 μmol/L HO-1 for 2 hours followed by 0.5 mmol/L H202）, HO-1 inhibition group [D group, pretreatment with l0 txmol/L zinc protoporphyrin IX （ZnppIX） for 2 hours followed by 0.5 mmol/L H202]. The ceils in each group were cuhm＇ed after different treatment. The cell apoptosis rate was determined by flow eytometry at 2, 6, 12 hours after the intervention. The protein expression of caspase-3 and Cyt-C were determined by Western Blot. Results The cell apoptosis rate in each group was gradually increased with prolonged time of H202 treatment. The apoptosis rate at different time points after treatment in B group was significantly higher than that in A group, while the rate in C group was significantly lower than that in B group [2 hours： （ 11.46 _＋ 1.47 ）% vs. （20.83 _＋ 1.55 ）%, 6 hours： （ 12.30 ± 1.37 ）% vs. （27.14 ± 1.53）%, 12 hours： （12.62 ± 1.39）% vs. （35.66 ＋ 0.74）%, all P〈0.05]. There were no significant differences in apoptotic rate at 2, 6, 12 hours between D group [（24.33 ± 1.36）%, （31.67 ± 1.24）%, （36.93 ± 2.40）% ] and B group. The protein expression of caspase-3 and Cyt-C was gradually increased with prolonged time of H202 treatment. The protein expression of easpase-3 and Cyt-C at different time points in B group was significantly higher than that in A group. The protein expression of caspase-3 （ gray scale ） in C group was significantly lower than thatin B group （2 ho

关 键 词：血红素加氧酶-1 凋亡 肺泡上皮细胞 Ⅱ型 过氧化氢 大鼠 

分 类 号：R563.8[医药卫生—呼吸系统]