CAJ | 학술논문

目的探讨电针足三里穴和内关穴对兔内毒素休克诱发心肌损伤的影响及血红素氧合酶-1(HO-1)在其中的作用.方法健康清洁级雄性新西兰大白兔60只,体重1.5～2.0 kg,2月龄,采用随机数字表法,将其分为6组(n=10):假手术组(S组)、锌原卟啉-Ⅸ组(Z组)、内毒素休克组(ES组)、电针+内毒素休克组(EES组)、假针刺+内毒素休克组(SEES组)、电针+内毒素休克+锌原卟啉-Ⅸ组(EESZ组).ES组、EES组、SEES组和EESZ组采用静脉注射脂多糖5 mg/kg的方法制备内毒素休克模型,S组和Z组给予等量生理盐水.EES组和EESZ组于模型制备前1～4d和模型制备当天(静脉注射内毒素开始至模型制备结束)电针双侧足三里穴和内关穴.疏密波,频率2/100 Hz,波宽0.2 ～ 0.6 ms,刺激强度2～3 mA,以免出现轻微肌颤为宜,30 min/次,1次/d.SEES组穴点定位于足三里穴和内关穴旁开0.5 cm处,刺激方法同EES组.EESZ组和Z组静脉注射脂多糖或生理盐水后2h时腹腔注射锌原卟啉-Ⅸ10 μmol/kg(溶于50 mmol/L的NaHCO3溶液1ml),其余组给予等容量NaHCO3溶液.静脉注射脂多糖或生理盐水后6h时,取右颈总动脉血样,测定血清TNF-α、CK和LDH水平,处死动物后取心肌组织,进行病理学评分;采用Western blot法测定HO-1蛋白的表达;采用荧光定量PCR法测定HO-1 mRNA的表达.结果与S组比较,ES组、EES组、SEES组、EESZ组心肌病理学评分、血清TNF-α、CK和LDH水平升高,心肌组织HO-1 mRNA及其蛋白表达上调(P＜0.05),Z组上述指标差异无统计学意义(P＞0.05);与ES组比较,EES组心肌病理学评分、血清TNF-α、CK、LDH水平降低,心肌组织HO-1 mRNA及其蛋白表达上调(P＜0.05),SEES组和EESZ组上述各指标差异无统计学意义(P＞0.05);与EES组比较,EESZ组心肌组织病理学评分、血清TNF-α、CK和LDH水平升高,心肌组织HO-1 mRNA及其蛋白表达下调(P＜0.05).结论电针足三里穴和内关穴可减轻内毒素休克诱发的兔心肌损伤,机制可能与上调心肌组织HO-1表达,抑制炎症反应有关. 目的探討電針足三裏穴和內關穴對兔內毒素休剋誘髮心肌損傷的影響及血紅素氧閤酶-1(HO-1)在其中的作用.方法健康清潔級雄性新西蘭大白兔60隻,體重1.5～2.0 kg,2月齡,採用隨機數字錶法,將其分為6組(n=10):假手術組(S組)、鋅原卟啉-Ⅸ組(Z組)、內毒素休剋組(ES組)、電針+內毒素休剋組(EES組)、假針刺+內毒素休剋組(SEES組)、電針+內毒素休剋+鋅原卟啉-Ⅸ組(EESZ組).ES組、EES組、SEES組和EESZ組採用靜脈註射脂多糖5 mg/kg的方法製備內毒素休剋模型,S組和Z組給予等量生理鹽水.EES組和EESZ組于模型製備前1～4d和模型製備噹天(靜脈註射內毒素開始至模型製備結束)電針雙側足三裏穴和內關穴.疏密波,頻率2/100 Hz,波寬0.2 ～ 0.6 ms,刺激彊度2～3 mA,以免齣現輕微肌顫為宜,30 min/次,1次/d.SEES組穴點定位于足三裏穴和內關穴徬開0.5 cm處,刺激方法同EES組.EESZ組和Z組靜脈註射脂多糖或生理鹽水後2h時腹腔註射鋅原卟啉-Ⅸ10 μmol/kg(溶于50 mmol/L的NaHCO3溶液1ml),其餘組給予等容量NaHCO3溶液.靜脈註射脂多糖或生理鹽水後6h時,取右頸總動脈血樣,測定血清TNF-α、CK和LDH水平,處死動物後取心肌組織,進行病理學評分;採用Western blot法測定HO-1蛋白的錶達;採用熒光定量PCR法測定HO-1 mRNA的錶達.結果與S組比較,ES組、EES組、SEES組、EESZ組心肌病理學評分、血清TNF-α、CK和LDH水平升高,心肌組織HO-1 mRNA及其蛋白錶達上調(P＜0.05),Z組上述指標差異無統計學意義(P＞0.05);與ES組比較,EES組心肌病理學評分、血清TNF-α、CK、LDH水平降低,心肌組織HO-1 mRNA及其蛋白錶達上調(P＜0.05),SEES組和EESZ組上述各指標差異無統計學意義(P＞0.05);與EES組比較,EESZ組心肌組織病理學評分、血清TNF-α、CK和LDH水平升高,心肌組織HO-1 mRNA及其蛋白錶達下調(P＜0.05).結論電針足三裏穴和內關穴可減輕內毒素休剋誘髮的兔心肌損傷,機製可能與上調心肌組織HO-1錶達,抑製炎癥反應有關.
목적 탐토전침족삼리혈화내관혈대토내독소휴극유발심기손상적영향급혈홍소양합매-1(HO-1)재기중적작용.방법 건강청길급웅성신서란대백토60지,체중1.5～2.0 kg,2월령,채용수궤수자표법,장기분위6조(n=10):가수술조(S조)、자원계람-Ⅸ조(Z조)、내독소휴극조(ES조)、전침+내독소휴극조(EES조)、가침자+내독소휴극조(SEES조)、전침+내독소휴극+자원계람-Ⅸ조(EESZ조).ES조、EES조、SEES조화EESZ조채용정맥주사지다당5 mg/kg적방법제비내독소휴극모형,S조화Z조급여등량생리염수.EES조화EESZ조우모형제비전1～4d화모형제비당천(정맥주사내독소개시지모형제비결속)전침쌍측족삼리혈화내관혈.소밀파,빈솔2/100 Hz,파관0.2 ～ 0.6 ms,자격강도2～3 mA,이면출현경미기전위의,30 min/차,1차/d.SEES조혈점정위우족삼리혈화내관혈방개0.5 cm처,자격방법동EES조.EESZ조화Z조정맥주사지다당혹생리염수후2h시복강주사자원계람-Ⅸ10 μmol/kg(용우50 mmol/L적NaHCO3용액1ml),기여조급여등용량NaHCO3용액.정맥주사지다당혹생리염수후6h시,취우경총동맥혈양,측정혈청TNF-α、CK화LDH수평,처사동물후취심기조직,진행병이학평분;채용Western blot법측정HO-1단백적표체;채용형광정량PCR법측정HO-1 mRNA적표체.결과 여S조비교,ES조、EES조、SEES조、EESZ조심기병이학평분、혈청TNF-α、CK화LDH수평승고,심기조직HO-1 mRNA급기단백표체상조(P＜0.05),Z조상술지표차이무통계학의의(P＞0.05);여ES조비교,EES조심기병이학평분、혈청TNF-α、CK、LDH수평강저,심기조직HO-1 mRNA급기단백표체상조(P＜0.05),SEES조화EESZ조상술각지표차이무통계학의의(P＞0.05);여EES조비교,EESZ조심기조직병이학평분、혈청TNF-α、CK화LDH수평승고,심기조직HO-1 mRNA급기단백표체하조(P＜0.05).결론 전침족삼리혈화내관혈가감경내독소휴극유발적토심기손상,궤제가능여상조심기조직HO-1표체,억제염증반응유관.
Objective To evaluate the effect of electro-acupuncture (EA) at Zusanli (ST-36) and Neiguan (PC6) acupoints on endotoxic shock (ES)-induced myocardial injury in rabbits and the role of heme oxygenase-1 (HO-1).Methods Sixty healthy male New Zealand white rabbits,aged 2 months,weighing 1.5-2.0 kg,were randomly assigned into 6 groups (n =10 each) using a random number table:sham operation group (group S) ; heme oxygenase-1 inhibitor zinc protoporphyrin-Ⅸ (ZnPP-Ⅸ) group (group Z) ; group ES; EA + ES group (group EES); sham EA + ES group (group SEES); EA + ES + ZnPP-Ⅸ group (group EESZ).The rabbits were anesthetized with intravenous 20％ urethane 5 ml/kg.Right common carotid artery was cannulated for BP monitoring.Ear vein was cannulated for drug administration.ES was induced with lipopolysaccharide (LPS) 5 mg/kg injected intravenously in ES,EES,SEES and EESZ groups,while the equal volume of normal saline was given in S and Z groups.ES was confirmed by decrease in MAP to 75 ％ of the baseline value.ZnPP-Ⅸ 10 μmol/kg (in 1 ml of NaHCO3 50 mmol/L) was injected intraperitoneally at 2 h after LPS or normal saline administration in EESZ and Z groups,while the equal volume of NaHCO3 was injected intraperitoneally in the other groups.In EES and EESZ groups,bilateral 30 min EA stimulation (0.2-0.6 ms,2/100 Hz,2-3 mA) of ST36 and PC6 was performed once a day on days 1-4 before induction of ES and on the day of induction of ES (from the onset of LPS injection until the end of induction of ES).In SEES group,electro-stimulation was performed at the points 0.5 cm lateral to the acupoints of ST36 and PC6 and the method was similar to those previously described in group EES.Blood samples were taken from the right common carotid artery at 6 h after LPS or normal saline administration for measurement of serum tumor necrosis factor-alpha (TNF-α),creatine kinase (CK),and lactic dehydrogenase (LDH) levels.Then the rabbits were sacrificed by exsanguination,and the myocardial tissues were removed for microscopic examination of the pathological changes which were scored and for determination of HO-1 protein (by Western blot) and mRNA (using fluorescence quantitative PCR) expression.Results Compared with group S,the pathological scores and concentrations of serum TNF-α,CK,and LDH were significantly increased,and the expression of HO-1 protein and mRNA was up-regulated in ES,EES,SEES and EESZ groups (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group Z (P ＞ 0.05).Compared with group ES,the pathological scores and concentrations of serum TNF-α,CK,and LDH were significantly decreased,and the expression of HO-1 protein and mRNA was up-regulated in group EES (P ＜ 0.05),while no significant change in the parameters mentioned above was found in SEES and EESZ groups (P ＞ 0.05).Compared with group EES,the pathological scores and concentrations of serum TNF-α,CK,and LDH were significantly increased,and the expression of HO-1 protein and mRNA was down-regulated in group EESZ (P ＜ 0.05).Conclusion EA at ST-36 and PC6 acupoints can attenuate ES-induced myocardial injury in rabbits and upregulation of HO-1 expression and inhibition of inflammatory responses may be involved in the mechanism.