摘要
目的探讨丁苯酞对局灶性脑缺血再灌注大鼠的脑保护机制。方法 160只雄性Sprague-Dawley大鼠随机分为假手术组(Sham组,n=10)、缺血再灌注组(IR组,n=50)、丁苯酞高剂量后处理组(高剂量组,n=50)、丁苯酞中剂量后处理组(中剂量组,n=25)和丁苯酞低剂量后处理组(低剂量组,n=25)。后4组线栓法制作缺血2 h再灌注模型。Sham组术后24 h处死,其他各组分别于再灌注6 h、12 h、24 h、48 h、72 h处死,5只用于应用原位末端标记(TUNEL)法检测细胞凋亡,免疫组织化学染色法观察沉默信息调节因子2相关酶1(SIRT1)及过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)的表达。Sham组、IR组及高剂量组另5只用实时荧光定量PCR检测SIRT1及PGC-1α的表达。结果与IR组比较,丁苯酞各剂量组各时间点凋亡细胞数均显著减少(F>160.60,P<0.001),SIRT1、PGC-1α阳性细胞数显著增多(F>87.20,P<0.001)。与低、中剂量组比较,高剂量组各时间点凋亡细胞数更少(P<0.05),SIRT1、PGC-1α阳性细胞数更多(P<0.05),低、中剂量组之间除再灌注6 h外,也有显著性差异(P<0.05)。与IR组比较,高剂量组各时间点SIRT1及PGC-1αm RNA表达均明显增多(t>4.13,P<0.01)。结论丁苯酞能抑制大鼠脑缺血再灌注后细胞凋亡,可能与上调SIRT1及PGC-1α的表达有关。
Objective To observe the neural protection of 3-n-butylphthalide（NBP） injection in focal cerebral ischemia-reperfusion rats.Methods 160 male Sprague-Dawley rats were randomly divided into sham group（n=10）,ischemia-reperfusion group（IR group,n=50）,high-dose NBP treatment group（high-dose group,n=50）,middle-dose NBP treatment group（middle-dose group,n=25） and low-dose NBP treatment group（low-dose group,n=25）.The later 4 groups were occluded the middle cerebral artery for 2 hours and reperfused.The sham group was sacrificed 24 hours after operation,and the other groups at 6,12,24,48 and 72 hours after reperfusion,in which 5 of them were stained with Td T mediated d UTP Nick End Labeling（TUNEL） to observe the neuronal apoptosis,and immunohistochemistry to observe the expression of silent information regulation 2 homolog 1（SIRT1） and peroxisome proliferator-activated receptor γ coactivator-1α（PGC-1α）;the other 5 of sham group,IR group and high-dose group were observed with quantitative real-time PCR of SIRT1 and PGC-1α.Results Compared with the IR group,the number of apoptotic cells decreased and the SIRT1 and PGC-1α positive cells increased in all NBP groups at each time（F〉160.60,P〈0.001）,and it was the least of apoptotic cells and most of SIRT1 and PGC-1α positive cells in the high-dose group（P〈0.05）,while there was significant difference between the low-dose group and the middle-dose group,excluding 6 hours after reperfusion（P〈0.05）.Compared with IR group,the expression of SIRT1 and PGC-1α m RNA increased in the high-dose group at each time（t〉4.13,P〈0.01）.Conclusion NBP can protect brain from apoptosis in focal cerebral ischemia-reperfusion rats,which may relate to more expression of SIRT1 and PGC-1α.
出处
《中国康复理论与实践》
CSCD
北大核心
2016年第1期32-37,共6页
Chinese Journal of Rehabilitation Theory and Practice
基金
河北省2013年医学科学研究重点课题计划(No.20130382)
