摘要
目的:观察应用硝普钠和一氧化氮合酶(NOS)抑制剂L-NAME对大鼠局灶性脑缺血再灌注所致神经细胞损伤的影响。方法:取SD大鼠44只,按随机数字表法分为4组:假手术组,脑缺血再灌注组,脑缺血再灌注加侧脑室微量注射L-NAME组,脑缺血再灌注加侧脑室微量注射硝普钠组。用栓线法复制大鼠大脑中动脉脑缺血再灌注模型,自颈内、外动脉分叉处起始,假手术组的尼龙栓子插入13mm,其余3组插入(18.0±0.5)mm造成大脑中动脉完全缺血30min,缓慢拔出尼龙栓子形成再灌注状态,L-NAME组和硝普钠组侧脑室微量注射L-NAME和硝普钠进行干预,假手术组和脑缺血再灌注组侧脑室注射等量生理盐水。于再灌注术后12,24h,2,4d分别处死动物取材,紫外分光光度计检测各脑组织一氧化氮含量,免疫组化法测脑组织NOS活性,TUNEL法检测调亡细胞。结果:脑缺血再灌注急性期(术后12h),脑组织一氧化氮含量迅速增高犤(10.14±1.97)mol/g犦,L-NAME明显降低脑组织一氧化氮的含量犤(3.86±1.35)mol/g犦,硝普钠能明显增加脑组织一氧化氮的含量犤(12.46±1.57)mol/g犦,SPSS10.0统计分析软件LSD法统计结果,各组间比较,F=24.07,P<0.05,有明显显著性意义。术后12hL-NAME抑制NOS的活性犤(16.0±1.2)个/视野犦,硝普钠组NOS的表达增强犤(62.0±4.2)个/视野犦,组间比较。
AIM:To observe the effects of sodium nitroprusside(SNP) and nitricoxide synthase(NOS) inhibitor L NAME on neural cell injury caused by focal ischemia reperfusion in rats. METHODS:Forty four SD rats were equally divided into four groups at random: sham operation group,ischemia reperfusion group(model group), ischemia reperfusion group with microinjection of L NAME into lateral ventricle(L NAME group),and ischemia reperfusion group with microinjection of SNP into lateral ventricle(SNP group).Rat models of cerebral ischemia reperfusion were made by middle cerebral artery occlusion(MACO) with nylon thread inserted at the crotch between internal and external carotid artery 13 mm in depth for the sham operation group and(18.0±0.5) mm for the other three groups.L NAME and SNP groups were treated with the microinjection of L NAME and SNP into lateral ventricle respectively.Sham operation group and model group were injected with isometric saline into lateral ventricle.The rats were killed respectively at 12 hours,24 hours, 2 days and 4 days after operation, the content of nitrogen monoxide(NO) in the brain was measured with ultraviolet spectrophotometer.The activity of NOS in the brain was detected with immunohistochemical method,and cell apoptosis was observed with TUNEL technique. RESULTS:At acute stage of ischemia reperfusion(12 hours after operation),the content of NO in the brain increased to(10.14±1.97) mol/g rapidly.L NAME decreased the content of NO to(3.86±1.35) mol/g, while SNP increased the content of NO to(12.46±1.57) mol/g.SPSS 10.0 software was used for statistical analysis with LSD method,and the results showed that there was significant difference between the two groups(F=24.07,P< 0.05).Twelve hours after the operation,L NAME inhibited the activity of NOS[(16.0±1.2) per visual field],while SNP activated the activity of NOS[(62.0±4.2) per visual field],showing a significant difference(F=21.3,P=0.003).Twenty four hours after operation, L-NAME could further inhibit the activity of NOS,while there was a higher expression of NOS in the SNP group, with significant difference between the two groups(F=16.18,P=0.023).Two days later,there were no significant changes in the activity of NOS between all the groups.TUNEL showed that the number of apoptotic cells increased in the L NAME group,and decreased in the SNP group.There was a significant difference between the two groups(F=12.3,P=0.019). CONCLUSION:NO can remarkably inhibit the apoptosis of nerve cells.
出处
《中国临床康复》
CSCD
2004年第10期1860-1862,共3页
Chinese Journal of Clinical Rehabilitation
