摘要
目的:观察肢体缺血再灌注所致脑损伤大鼠脑组织中一氧化氮的变化,分析一氧化氮在损伤中的作用及MK801对其的影响。方法:实验于2003-09/2004-12在华北煤炭医学院解剖教研室和实验中心完成。①40只Wistar大鼠随机分成5组,每组8只:对照组、再灌4h组(缺血4h再灌4h)、再灌12h组(缺血4h再灌12h)、再灌24h组(缺血4h再灌24h),MK801干预组(缺血4h再灌24h+MK801)。②复制大鼠肢体缺血再灌损伤模型,MK801干预组在再灌注前5min立刻皮下注射MK801(0.5mg/kg)。③分别测定各组大鼠脑组织中一氧化氮、丙二醛含量、免疫组化观察各组脑组织诱导型一氧化氮合酶表达变化、并用Western-blot检测诱导型一氧化氮合酶蛋白表达。结果:40只大鼠均进入结果分析。①MK801干预组与再灌24h组比较,丙二醛含量降低了45.8%,一氧化氮含量降低了12.8%。②免疫组化结果显示随再灌时间的延长,大鼠脑组织诱导型一氧化氮合酶表达呈逐渐上升趋势,再灌24h达高峰。阳性信号主要分布海马区,中脑红核区等部位的神经元细胞,对照组仅见少量表达,而MK801干预后诱导型一氧化氮合酶表达明显降低,同时减轻脑组织水肿及神经元受损。与Western-blot检测诱导型一氧化氮合酶蛋白表达结果一致。结论:MK801减少脑组织一氧化氮、丙二醛含量,降低诱导型一氧化氮合酶表达,说明MK801可减轻肢体缺血再灌注所致脑损伤,可能与一氧化氮含量及诱导型一氧化氮合酶表达在肢体缺血再灌注所致脑损伤中降低有关。
AIM： To observe the changes of nitric oxide （NO） in the development of brain injury after limbs ischemia-reperfusion （LIR）, and the effect of NO in injury and the effect of MK801 on NO.
METHODS： The experiment was conducted at the Staff Room of Anatomy and Experimental Center of North China Coal Medical College Between September 2003 and December 2004. （1） Totally 40 Wistar rats were randomly divided into five groups with 8 rats in each group： control group, reperfusion 4-hour group （ischemia for 4 hours and reperfusion for 4 hours）, reperfusion 12-hour group （ischemia for 4 hours and reperfusion for 12 hours）, reperfusion 24-hour group （ischemia for 4 hours and reperfusion for 24 hours）, and MK801 interventional group （ischemia for 4 hours and reperfusion for 24 hours ＋ MK801）. （2）LIR models of rats were duplicated. Five minutes before reperfusion the rats in the MK801 interventional group were treated with MK801 （0.5 mg/kg） by subcutaneous injection immediately. （3）Contents of NO and malondialdehyde （MDA） in the brain tissue of rats of each group were detected, respectively. Change of expression of inducible nitric oxide synthase （iNOS） of brain tissue of each group was observed with immunohistochemical method, and protein expression of iNOS was detected with Western-blot.
RESULTS： Totally 40 rats were involved in the result analysis. （1） Compared between the MK801 interventional group and reperfusion 24- hour group, the content of MDA decreased by 45.8%, and the content of NO reduced by 12.8%. （2）The immunohistochemical result showed that the expression of iNOS of brain tissue of rats increased gradually with the prolongation of reperfusion duration, and reached the peak at the 24^th hour. Positive single mainly distributed at neurons of hippocampi and red nucleus of midbrain. There was slight expression in the control group, while the expression of iNOS after the intervention with MK801 reduced significantly; Meanwhile, the injuries of edema and neurons of brain tissue were relieved, which was the same to the result of Western-blot.
CONCLUSION： MK801 decreases the contents of NO and MDA, decreases the expression level of iNOS. It is indicated that MK801 may alleviate brain injury following LIR, which may be related with the decreasing the content of NO and the expression of iNOS following LIR.
出处
《中国临床康复》
CSCD
北大核心
2006年第28期102-104,共3页
Chinese Journal of Clinical Rehabilitation
