中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2014年
3期
348-352
,共5页
宫丽荣%余剑波%徐妍%曹新顺%史佳%张桂诚
宮麗榮%餘劍波%徐妍%曹新順%史佳%張桂誠
궁려영%여검파%서연%조신순%사가%장계성
转录因子AP-1%电刺激疗法%休克 ,脓毒性%肺%血红素加氧酶-1
轉錄因子AP-1%電刺激療法%休剋 ,膿毒性%肺%血紅素加氧酶-1
전록인자AP-1%전자격요법%휴극 ,농독성%폐%혈홍소가양매-1
Transcription factor AP-1%Electric stimulation therapy%Shock,septic%Lung%Heme oxygenase-1
目的：评价活化蛋白-1（AP-1）在电针上调内毒素休克兔肺组织血红素加氧酶-1（HO-1）表达中的作用。方法健康雄性新西兰大白兔70只，体重1.5～2.0 kg ，2月龄，采用随机数字表法，将其分为7组（ n＝10）：正常对照组（C组）、内毒素休克诱发急性肺损伤组（ALI组）、电针＋ALI组（EL组）、非穴位＋电针＋ALI组（NEL组）、电针＋ALI＋姜黄素组（ELC组）、AP-1抑制剂姜黄素组（Cur组）和二甲基亚砜组（D组）。EL组和ELC组电针刺激双侧足三里及肺俞穴，疏密波，频率15 Hz ，刺激强度以兔出现轻微肌颤为宜，15 min/次，1次/d ，连续5 d ，留针1 h后出针；NEL组以相同电针参数刺激足三里及肺俞穴旁开0.5 cm处。电针刺激5 d后，Cur组及ELC组经耳缘静脉注射姜黄素25 mg/kg （溶于0.5ml0.1％二甲基亚砜），D组静脉注射0.1％二甲基亚砜0.5ml，其余组给予等容量的生理盐水。给药后30 min ALI组、EL组、NEL组及ELC组经耳缘静脉缓慢注射LPS 5 mg/kg （溶于2 ml生理盐水），其余3组给予等容量的生理盐水。给予LPS或生理盐水后6 h时处死取肺组织行病理学评分，计算肺湿/干重比值（W/D比值），检测肺组织MDA含量及SOD活性，采用Western blot法测定HO-1及c-Jun表达水平，采用荧光定量PCR法测定HO-1 mRNA及c-Jun mRNA表达水平。结果与C组比较，ALI组、EL组、NEL组及ELC组肺组织病理学评分、W/D比值和MDA含量升高，SOD活性降低，ALI组、EL组及NEL组HO-1 mRNA、HO-1、c-Jun mRNA和c-Jun表达上调（ P＜0.05），ELC组c-Jun mRNA和c-Jun表达水平差异无统计学意义（ P＞0.05），Cur组及D组上述各指标差异无统计学意义（ P＞0.05）。与ALI组比较，EL组及ELC组肺组织病理学评分、W/D比值和MDA含量降低，SOD活性升高，HO-1 mRNA和HO-1表达上调，EL组c-Jun mRNA和c-Jun表达上调，ELC组c-Jun mRNA和c-Jun表达下调（ P＜0.05）， NEL组上述各指标差异无统计学意义（ P＞0.05）；与EL组比较，ELC组肺组织病理学评分、W/D比值、MDA含量升高，SOD活性降低，HO-1 mRNA、HO-1、c-Jun mRNA及c-Jun表达下调（ P＜0.05）。结论内毒素休克诱发兔急性肺损伤时，电针可通过活化AP-1而上调HO-1表达起到肺保护作用。
目的：評價活化蛋白-1（AP-1）在電針上調內毒素休剋兔肺組織血紅素加氧酶-1（HO-1）錶達中的作用。方法健康雄性新西蘭大白兔70隻，體重1.5～2.0 kg ，2月齡，採用隨機數字錶法，將其分為7組（ n＝10）：正常對照組（C組）、內毒素休剋誘髮急性肺損傷組（ALI組）、電針＋ALI組（EL組）、非穴位＋電針＋ALI組（NEL組）、電針＋ALI＋薑黃素組（ELC組）、AP-1抑製劑薑黃素組（Cur組）和二甲基亞砜組（D組）。EL組和ELC組電針刺激雙側足三裏及肺俞穴，疏密波，頻率15 Hz ，刺激彊度以兔齣現輕微肌顫為宜，15 min/次，1次/d ，連續5 d ，留針1 h後齣針；NEL組以相同電針參數刺激足三裏及肺俞穴徬開0.5 cm處。電針刺激5 d後，Cur組及ELC組經耳緣靜脈註射薑黃素25 mg/kg （溶于0.5ml0.1％二甲基亞砜），D組靜脈註射0.1％二甲基亞砜0.5ml，其餘組給予等容量的生理鹽水。給藥後30 min ALI組、EL組、NEL組及ELC組經耳緣靜脈緩慢註射LPS 5 mg/kg （溶于2 ml生理鹽水），其餘3組給予等容量的生理鹽水。給予LPS或生理鹽水後6 h時處死取肺組織行病理學評分，計算肺濕/榦重比值（W/D比值），檢測肺組織MDA含量及SOD活性，採用Western blot法測定HO-1及c-Jun錶達水平，採用熒光定量PCR法測定HO-1 mRNA及c-Jun mRNA錶達水平。結果與C組比較，ALI組、EL組、NEL組及ELC組肺組織病理學評分、W/D比值和MDA含量升高，SOD活性降低，ALI組、EL組及NEL組HO-1 mRNA、HO-1、c-Jun mRNA和c-Jun錶達上調（ P＜0.05），ELC組c-Jun mRNA和c-Jun錶達水平差異無統計學意義（ P＞0.05），Cur組及D組上述各指標差異無統計學意義（ P＞0.05）。與ALI組比較，EL組及ELC組肺組織病理學評分、W/D比值和MDA含量降低，SOD活性升高，HO-1 mRNA和HO-1錶達上調，EL組c-Jun mRNA和c-Jun錶達上調，ELC組c-Jun mRNA和c-Jun錶達下調（ P＜0.05）， NEL組上述各指標差異無統計學意義（ P＞0.05）；與EL組比較，ELC組肺組織病理學評分、W/D比值、MDA含量升高，SOD活性降低，HO-1 mRNA、HO-1、c-Jun mRNA及c-Jun錶達下調（ P＜0.05）。結論內毒素休剋誘髮兔急性肺損傷時，電針可通過活化AP-1而上調HO-1錶達起到肺保護作用。
목적：평개활화단백-1（AP-1）재전침상조내독소휴극토폐조직혈홍소가양매-1（HO-1）표체중적작용。방법건강웅성신서란대백토70지，체중1.5～2.0 kg ，2월령，채용수궤수자표법，장기분위7조（ n＝10）：정상대조조（C조）、내독소휴극유발급성폐손상조（ALI조）、전침＋ALI조（EL조）、비혈위＋전침＋ALI조（NEL조）、전침＋ALI＋강황소조（ELC조）、AP-1억제제강황소조（Cur조）화이갑기아풍조（D조）。EL조화ELC조전침자격쌍측족삼리급폐유혈，소밀파，빈솔15 Hz ，자격강도이토출현경미기전위의，15 min/차，1차/d ，련속5 d ，류침1 h후출침；NEL조이상동전침삼수자격족삼리급폐유혈방개0.5 cm처。전침자격5 d후，Cur조급ELC조경이연정맥주사강황소25 mg/kg （용우0.5ml0.1％이갑기아풍），D조정맥주사0.1％이갑기아풍0.5ml，기여조급여등용량적생리염수。급약후30 min ALI조、EL조、NEL조급ELC조경이연정맥완만주사LPS 5 mg/kg （용우2 ml생리염수），기여3조급여등용량적생리염수。급여LPS혹생리염수후6 h시처사취폐조직행병이학평분，계산폐습/간중비치（W/D비치），검측폐조직MDA함량급SOD활성，채용Western blot법측정HO-1급c-Jun표체수평，채용형광정량PCR법측정HO-1 mRNA급c-Jun mRNA표체수평。결과여C조비교，ALI조、EL조、NEL조급ELC조폐조직병이학평분、W/D비치화MDA함량승고，SOD활성강저，ALI조、EL조급NEL조HO-1 mRNA、HO-1、c-Jun mRNA화c-Jun표체상조（ P＜0.05），ELC조c-Jun mRNA화c-Jun표체수평차이무통계학의의（ P＞0.05），Cur조급D조상술각지표차이무통계학의의（ P＞0.05）。여ALI조비교，EL조급ELC조폐조직병이학평분、W/D비치화MDA함량강저，SOD활성승고，HO-1 mRNA화HO-1표체상조，EL조c-Jun mRNA화c-Jun표체상조，ELC조c-Jun mRNA화c-Jun표체하조（ P＜0.05）， NEL조상술각지표차이무통계학의의（ P＞0.05）；여EL조비교，ELC조폐조직병이학평분、W/D비치、MDA함량승고，SOD활성강저，HO-1 mRNA、HO-1、c-Jun mRNA급c-Jun표체하조（ P＜0.05）。결론내독소휴극유발토급성폐손상시，전침가통과활화AP-1이상조HO-1표체기도폐보호작용。
Objective To evaluate the role of activator protein-1 (AP-1 ) in electro-acupuncture (EA )-induced up-regulation of heme oxygenase-1 (HO-1 ) expression in lung tissues in a rabbit model of endotoxic shock .Methods Seventy healthy male New Zealand white rabbits ,aged 2 months ,weighing 1.5-2.0 kg ,were randomly divided into 7 groups ( n=10 each ) using a random number table :normal control group (group C ) , endotoxic shock-induced acute lung injury (ALI ) group (group ALI ) , EA + ALI group (group EL ) , non-acupoint+EA+ ALI group (group NEL ) , curcumin (HO-1 inhibitor ) group (group Cur ) , dimethyl sulfoxide group (group D ) ,and EA+ALI+curcumin group (group ELC ) .Bilateral 15 min EA stimulation of Zusanli and Feishu (according to atlas of animals acupoints ) was performed (frequency 15Hz ) once a day for 5 consecutive days before lipopolysaccharide (LPS ) administration in EL and ELC groups ,while in group NEL ,EA stimulation was performed at non-acupoints located 0.5 cm lateral to Zusanli and Feishu acupoints with the same parameters . The animals were anesthetized with urethane and tracheostomized .The animals kept spontaneous breathing .Right internal carotid artery was cannulated for blood pressure monitoring . Ear vein was cannulated for drug administration .At 5 days after EA stimulation ,curcumin 25 mg/kg (in 0.5 ml of 0.1% dimethyl sulfoxide ) was injected in Cur and ELC groups ,0.1% dimethyl sulfoxide 0.5 ml was injected in D group ,and the equal volume of normal saline was given in the other groups .LPS 5 mg/kg (in 2 ml of 0.9% normal saline ) was injected intravenously at 30 min after administration in ALI ,EL ,NEL and ELC groups ,while the equal volume of normal saline was given in the other three groups .Endotoxic shock was confirmed by decrease in mean arterial pressure to 75% of the baseline value within 2 h after LPS injection .Blood samples were collected from the common carotid artery at 6 h after LPS or normal saline administration and the rabbits were then sacrificed .The lungs were removed for microscopic examination and the pathological changes were scored .The wet/dry lung weight ratio (W/D ratio ) was calculated .The malondialdehyde (MDA ) content and superoxide dismutase (SOD ) activity in lung tissues were detected ,and the expression of HO-1 mRNA ,HO-1 ,c-Jun mRNA and c-Jun in lung tissues was determined by Western blot .Results Compared with group C ,the pathological score ,W/D ratio and MDA content were significantly increased ,and the SOD activity was decreased in ALI ,EL ,NEL and ELC groups ,the expression of HO-1 mRNA ,HO-1 ,c-Jun mRNA and c-Jun was up-regulated in groups ALI ,EL and NEL ( P<0.05 ) ,while no significant change was found in the expression of c-Jun mRNA and c-Jun in group ELC ( P>0.05) .There was no significant difference in the parameters mentioned above between Cur and D groups ( P>0.05 ) .Compared with group ALI ,the pathological score ,W/D ratio and MDA content were significantly decreased ,and SOD activity was increased ,and the expression of HO-1 mRNA and HO-1 was up-regulated in EL and ELC groups ( P<0.05) ,the expression of c-Jun mRNA and c-Jun was up-regulated in group EL ,while down-regulated in group ELC ( P<0.05) ,and no significant change was found in the parameters mentioned above in group NEL ( P>0.05) .The pathological score ,W/D ratio and MDA content were significantly higher ,and the SOD activity and expression of HO-1 mRNA ,HO-1 ,c-Jun mRNA and c-Jun were lower in group ELC than in group EL ( P<0.05) .Conclusion EA up-regulates HO-1 expression through activating AP-1 during endotoxic shock-induced ALI in rabbits ,thus protecting the lung .
