CAJ | 학술논문

目的探讨电针减轻内毒素休克诱发兔急性肺损伤的机制与核因子E2相关因子2/抗氧化反应元件(Nrf2/ARE)通路的关系.方法健康雄性新西兰大白兔40只,2月龄,体重1.5～2.0kg,采取随机数字表法,将其分为4组(n=10):假手术组(S组)、内毒素休克组(ES组)、电针刺激穴位+内毒素休克组(EA组)、电针刺激非穴位+内毒素休克组(NA-EA组).ES组、EA组及NA-EA组静脉注射脂多糖(LPS)5 mg/kg制备内毒素休克模型.S组给予等容量的生理盐水.EA组于模型制备前1～4d及模型制备过程中电针刺激双侧足三里、内关和肺俞穴(疏密波,频率2/15 Hz,刺激电流1～2 mA,波宽0.2 ～ 0.6 ms,刺激强度以兔肢体出现轻微颤动为宜),1次/d,30 min/次;NA-EA组电针刺激足三里、内关和肺俞穴旁开0.5 cm非经非穴处,针刺参数同EA组.静脉注射LPS或生理盐水后6h时,处死动物,取肺组织,进行病理学观察及肺损伤评分,计算肺组织湿重/干重(W/D)比值,测定肺组织MDA含量及SOD活性,检测Nrf2 mRNA、核蛋白和总蛋白Nrf2的表达水平,计算核蛋白与总蛋白Nrf2表达水平的比值(N/T比值).结果与S组比较,ES组、EA组和NA-EA组肺组织W/D比值、MDA含量和肺组织损伤评分升高,SOD活性降低,肺组织Nrf2 mRNA、总蛋白及核蛋白Nrf2表达上调,N/T比值升高(P＜0.05);与ES组比较,EA组肺组织W/D比值、MDA含量和肺组织损伤评分降低,SOD活性升高,肺组织Nrf2 mRNA、总蛋白及核蛋白Nr2表达上调,N/T比值升高(P＜0.05),NA-EA组上述指标差异无统计学意义(P＞0.05).结论电针减轻内毒素休克诱发兔急性肺损伤的机制可能与激活Nrf2/ARE通路,增强抗氧化能力有关.
목적 탐토전침감경내독소휴극유발토급성폐손상적궤제여핵인자E2상관인자2/항양화반응원건(Nrf2/ARE)통로적관계.방법 건강웅성신서란대백토40지,2월령,체중1.5～2.0kg,채취수궤수자표법,장기분위4조(n=10):가수술조(S조)、내독소휴극조(ES조)、전침자격혈위+내독소휴극조(EA조)、전침자격비혈위+내독소휴극조(NA-EA조).ES조、EA조급NA-EA조정맥주사지다당(LPS)5 mg/kg제비내독소휴극모형.S조급여등용량적생리염수.EA조우모형제비전1～4d급모형제비과정중전침자격쌍측족삼리、내관화폐유혈(소밀파,빈솔2/15 Hz,자격전류1～2 mA,파관0.2 ～ 0.6 ms,자격강도이토지체출현경미전동위의),1차/d,30 min/차;NA-EA조전침자격족삼리、내관화폐유혈방개0.5 cm비경비혈처,침자삼수동EA조.정맥주사LPS혹생리염수후6h시,처사동물,취폐조직,진행병이학관찰급폐손상평분,계산폐조직습중/간중(W/D)비치,측정폐조직MDA함량급SOD활성,검측Nrf2 mRNA、핵단백화총단백Nrf2적표체수평,계산핵단백여총단백Nrf2표체수평적비치(N/T비치).결과 여S조비교,ES조、EA조화NA-EA조폐조직W/D비치、MDA함량화폐조직손상평분승고,SOD활성강저,폐조직Nrf2 mRNA、총단백급핵단백Nrf2표체상조,N/T비치승고(P＜0.05);여ES조비교,EA조폐조직W/D비치、MDA함량화폐조직손상평분강저,SOD활성승고,폐조직Nrf2 mRNA、총단백급핵단백Nr2표체상조,N/T비치승고(P＜0.05),NA-EA조상술지표차이무통계학의의(P＞0.05).결론 전침감경내독소휴극유발토급성폐손상적궤제가능여격활Nrf2/ARE통로,증강항양화능력유관.
Objective To investigate the relationship between the mechanism of electroacupuncture (EA)-induced reduction of acute lung injury induced by endotoxic shock (ES) and nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway in rabbits.Methods Forty healthy male New Zealand white rabbits,aged 2 months,weighing 1.5-2.0 kg,were randomly divided into 4 groups (n =10 each) using a random number table:sham operation group (group S),group ES,EA at acupoints + ES group (group EA),and EA at non-acupoints + ES group (group NA-EA).The animals were anesthetized with iv 20％ urethane 5 ml/kg,tracheostomized and kept spontaneous breathing.ES was induced by iv administration of lipopolysaccharide (LPS) 5 mg/kg in ES,EA and NA-EA groups.The equal volume of normal saline was given in group S.Bilateral 30 min EA (wave length 0.2-0.6 ms,frequency 2/100 Hz,intensity ≤2-3 mA) stimulation of Zusanli and Feishu was performed once a day for 4 days before establishment of endotoxic shock model and during the process of establishment of endotoxic shock model in EA group.In NA-EA group,EA was performed at the points 0.5 cm lateral to the acupoints of Zusanli and Feishu according to the method previously described in group EA.The rabbits were sacrificed at 6 h after LPS or normal saline administration,and lungs were removed for microscopic examination and for determination of wet to dry lung weight ratio (W/D ratio),malondialdehyde (MDA) contents,superoxide dismutase (SOD) activities,and expression of Nrf2 mRNA in lung tissues and Nrf2 in nucleoprotein and total protein.The pathological changes of lung were scored.The ratio of Nrf2 expression in nucleoprotein to Nrf2 expression in total protein (N/T ratio) was calculated.Results Compared with group S,W/D ratio,MDA contents,and lung injury scores were significantly increased,the activities of SOD were decreased,the expression of Nrf2 mRNA in lung tissues and Nrf2 in nucleoprotein and total protein was up-regulated,and N/T ratio was increased in ES,EA and NA-EA groups.Compared with group ES,W/D ratio,MDA contents,and lung injury scores were significantly decreased,the activities of SOD were increased,the expression of Nrf2 mRNA in lung tissues and Nrf2 in nucleoprotein and total protein was up-regulated,and N/T ratio was increased in EA group,and no significant changes were found in NA-EA group.Conclusion The mechanism by which EA reduces acute lung injury induced by ES may be related to activation of Nrf2/ARE pathway and enhancement of antioxidant capacity in rabbits.