中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2014年
12期
1481-1484
,共4页
戴琼艳%龙云%赵琳%张露%高宇%吴淋%陈兴东%段满林%徐建国
戴瓊豔%龍雲%趙琳%張露%高宇%吳淋%陳興東%段滿林%徐建國
대경염%룡운%조림%장로%고우%오림%진흥동%단만림%서건국
电压依赖性阴离子通道1%氢%再灌注损伤%脑
電壓依賴性陰離子通道1%氫%再灌註損傷%腦
전압의뢰성음리자통도1%경%재관주손상%뇌
Voltage-dependent anion channel 1%Hydrogen%Reperfusion injury%Brain
目的 评价电压依赖性阴离子通道1(VDAC1)在富氢液减轻大鼠全脑缺血再灌注损伤中的作用.方法 雄性SD大鼠60只,体重280 ~ 320 g,采用随机数字表法,将其分为6组(n=10):假手术组(S组)、缺血再灌注组(I/R组)、富氢液组(H组)、二甲基亚砜溶剂对照组(DMSO组)、VDAC1阻断剂4'-二异硫氰基芪-2,2'-二磺酸组(DIDS组)和富氢液+DIDS组(HD组).采用经食管心脏起搏诱发室颤后心肺复苏建立大鼠全脑缺血再灌注损伤模型.H组和HD组于再灌注即刻和6h时腹腔注射富氢液5 ml/kg,I/R组腹腔注射等容量生理盐水;DIDS组和HD组诱发室颤前经15 min股静脉输注DIDS 12 mg/kg,次日9时腹腔注射DIDS 12 mg/kg;DMSO组诱发室颤前经15 min股静脉输注DIDS的溶剂DMSO 12 mg/kg,次日9时腹腔注射DMSO 12 mg/kg.于再灌注24h时行神经功能评分,然后处死大鼠,取海马组织,计数正常椎体细胞,采用Western blot法测定VDAC1蛋白、细胞色素c和活化的caspase-3表达水平,采用荧光探针法测定活性氧(ROS)的含量.结果 与S组比较,其余各组神经功能评分和正常锥体细胞计数降低,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达上调,ROS含量升高(P<0.05);与I/R组比较,H组、DIDS组和HD组神经功能评分和正常锥体细胞计数升高,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达下调,ROS含量降低(P<0.05),DMSO组上述各指标差异无统计学意义(P>0.05);与H组比较,HD组神经功能评分和正常锥体细胞计数升高,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达下调,ROS含量降低(P<0.05).结论 富氢液通过抑制VDAC1的开放减轻大鼠全脑缺血再灌注损伤.
目的 評價電壓依賴性陰離子通道1(VDAC1)在富氫液減輕大鼠全腦缺血再灌註損傷中的作用.方法 雄性SD大鼠60隻,體重280 ~ 320 g,採用隨機數字錶法,將其分為6組(n=10):假手術組(S組)、缺血再灌註組(I/R組)、富氫液組(H組)、二甲基亞砜溶劑對照組(DMSO組)、VDAC1阻斷劑4'-二異硫氰基芪-2,2'-二磺痠組(DIDS組)和富氫液+DIDS組(HD組).採用經食管心髒起搏誘髮室顫後心肺複囌建立大鼠全腦缺血再灌註損傷模型.H組和HD組于再灌註即刻和6h時腹腔註射富氫液5 ml/kg,I/R組腹腔註射等容量生理鹽水;DIDS組和HD組誘髮室顫前經15 min股靜脈輸註DIDS 12 mg/kg,次日9時腹腔註射DIDS 12 mg/kg;DMSO組誘髮室顫前經15 min股靜脈輸註DIDS的溶劑DMSO 12 mg/kg,次日9時腹腔註射DMSO 12 mg/kg.于再灌註24h時行神經功能評分,然後處死大鼠,取海馬組織,計數正常椎體細胞,採用Western blot法測定VDAC1蛋白、細胞色素c和活化的caspase-3錶達水平,採用熒光探針法測定活性氧(ROS)的含量.結果 與S組比較,其餘各組神經功能評分和正常錐體細胞計數降低,海馬組織VDAC1蛋白、細胞色素c和活化的caspase-3錶達上調,ROS含量升高(P<0.05);與I/R組比較,H組、DIDS組和HD組神經功能評分和正常錐體細胞計數升高,海馬組織VDAC1蛋白、細胞色素c和活化的caspase-3錶達下調,ROS含量降低(P<0.05),DMSO組上述各指標差異無統計學意義(P>0.05);與H組比較,HD組神經功能評分和正常錐體細胞計數升高,海馬組織VDAC1蛋白、細胞色素c和活化的caspase-3錶達下調,ROS含量降低(P<0.05).結論 富氫液通過抑製VDAC1的開放減輕大鼠全腦缺血再灌註損傷.
목적 평개전압의뢰성음리자통도1(VDAC1)재부경액감경대서전뇌결혈재관주손상중적작용.방법 웅성SD대서60지,체중280 ~ 320 g,채용수궤수자표법,장기분위6조(n=10):가수술조(S조)、결혈재관주조(I/R조)、부경액조(H조)、이갑기아풍용제대조조(DMSO조)、VDAC1조단제4'-이이류청기기-2,2'-이광산조(DIDS조)화부경액+DIDS조(HD조).채용경식관심장기박유발실전후심폐복소건립대서전뇌결혈재관주손상모형.H조화HD조우재관주즉각화6h시복강주사부경액5 ml/kg,I/R조복강주사등용량생리염수;DIDS조화HD조유발실전전경15 min고정맥수주DIDS 12 mg/kg,차일9시복강주사DIDS 12 mg/kg;DMSO조유발실전전경15 min고정맥수주DIDS적용제DMSO 12 mg/kg,차일9시복강주사DMSO 12 mg/kg.우재관주24h시행신경공능평분,연후처사대서,취해마조직,계수정상추체세포,채용Western blot법측정VDAC1단백、세포색소c화활화적caspase-3표체수평,채용형광탐침법측정활성양(ROS)적함량.결과 여S조비교,기여각조신경공능평분화정상추체세포계수강저,해마조직VDAC1단백、세포색소c화활화적caspase-3표체상조,ROS함량승고(P<0.05);여I/R조비교,H조、DIDS조화HD조신경공능평분화정상추체세포계수승고,해마조직VDAC1단백、세포색소c화활화적caspase-3표체하조,ROS함량강저(P<0.05),DMSO조상술각지표차이무통계학의의(P>0.05);여H조비교,HD조신경공능평분화정상추체세포계수승고,해마조직VDAC1단백、세포색소c화활화적caspase-3표체하조,ROS함량강저(P<0.05).결론 부경액통과억제VDAC1적개방감경대서전뇌결혈재관주손상.
Objective To evaluate the role of voltnge-dependent anion channel 1 (VDAC1) in attenuation of global cerebral ischemia-reperfusion (I/R) injury by hydrogen-rich saline in rats.Methods Sixty male Sprague Dawley rats,weighing 280-320 g,were randomly divided into 6 groups (n =10 each) using a random number table:sham operation group (group S),I/R group,hydrogen-rich saline group (group H),vehicle dimethyl sulfoxide (DMSO) group,VDAC1 blocker 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) group (group DIDS),and hydrogen-rich saline + DIDS group (group HD).Ventricular fibrillation (VF) was induced by transoesophageal cardiac pacing and cardiopulmonary resuscitation was initiated 4 min after onset of VF to establish the model of global cerebral I/R injury.In H and HD groups,hydrogen-rich saline 5 ml/mg was injected intraperitoneally at 0 and 6 h after reperfusion,while the equal volume of normal saline was injected instead of hydrogen-rich saline in group I/R.In DIDS and HD groups,DIDS 12 mg/kg was infused over 15 min via the femoral vein prior to induction of VF,and DIDS 12 mg/kg was injected intraperitoneally at 9:00 am on the next day.In DMSO group,DMSO 12 mg/kg was infused over 15 min viathe femoral veprior to induction of VF,and DMSO 12 mg/kg was injected intraperitoneally at 9:00 am on the next day.After the neurological score was evaluated at 24 h of reperfusion,the rats were sacrificed.Hippocampi were immediately removed for determination of the number of normal pyramidal neurons,expression of VDAC1 protein,cytochrome c and activated caspase-3 (by Western blot),and content of reactive oxygen species (ROS) (by using fluorescence probe).Results Compared with group S,the neurological scores and the number of normal pyramidal neurons were significantly decreased,the expression of VDAC1 protein,cytochrome c and activated caspase-3 was up-regulated,and the content of ROS was increased in the other groups.Compared with group I/R,the neurologicalscores and the number of normal pyramidal neurons were significantly increased,the expression of VDAC1 protein,cytochrome c and activated caspase-3 was down-regulated,and the content of ROS was decreased in H,DIDS and HD groups,and no significant change was fotund in each parameter mentioned above in DMSO group.Compared with group H,the neurological scores and the number of normal pyramidal neurons were significantly increased,the expression of VDAC1 protein,cytochrome e and activated caspase-3 was down-regulated,and the content of ROS was decreased in HD group.Conclusion Hydrogen-richsaline attenuates global cerebral I/R injury through inhibiting VDAC1 opening in rats.