脑红蛋白在脑梗死大鼠表达及丁苯酞干预效应

Expression of neuroglobin on focal cerebral ischemia in rats and butylphthalide conditioning protection

目的探讨脑红蛋白在脑梗死大鼠中的表达和丁苯酞干预在氧化应激损伤中作用。方法将90只雄性SD大鼠随机分为假手术组、模型组、治疗组。每组再分为3个亚组:1 d组、3 d组和7 d组,每亚组10只。模型组和治疗组采用线栓法制作大鼠大脑中动脉闭塞模型(MCAO),假手术组只分离不结扎。治疗组在动物苏醒后以丁苯酞植物油灌胃,假手术组和模型组同法给予等量植物油灌胃。每只大鼠在术后3h和处死前行神经功能评分,应用RT-PCR和免疫组化检测脑组织脑红蛋白(NGB)表达,化学比色法检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果 (1)神经功能评分,模型组和治疗组术后3 h评分差异无统计学意义(P>0.05),各时间点处死前评分差异均有统计学意义(P<0.05)。(2)NGB mRNA和免疫组化表达随时间延长逐渐降低,各时间点模型组较假手术组、治疗组较模型组高,差异均有统计学意义(P<0.05)。(3)SOD活性和MDA含量随时间延长逐渐减少。SOD活性假手术组较治疗组、治疗组较模型组高,差异均有统计学意义(P<0.05);MDA含量假手术组较治疗组、治疗组较模型组低,差异均有统计学意义(P<0.05)。结论丁苯酞促进脑缺血大鼠脑红蛋白表达,减少氧化应激损伤。

Objective To investigate the expression of neuroglobin （NGB） on focal cerebral ischemia in rats and the effects of butylphthalide （NBP） conditioning via ameliorating oxidative stress injury. Methods Male SD rats （ n = 90） were randomly divided into three groups：sham, model and treatment group. Then each group was divided into three sub- groups：l d group,3d group and 7d group,each subgroup contained ten rats. Model and treatment groups was were operated reversible middle cerebral artery occlusion （MCAO） by intraluminal suture technique in rats and sham group was just sepa- rated the blood vessels. Treatment group was given butylphthalide vegetable oil by intragastric administration after recovery from anaesthesia, while sham and model groups was were given the same dose of vegetable oil. Each rat was estimated the nerve function defects at the time of 3 hours after operation and before being killed. The expression of NGB was determined by Real-time PCR and immuuohistochemistry, while superoxide dismutase （SOD） activity and malondialdehyde （MDA） content in brain tissue determined for superoxide dismutase （SOD） activity and malondialdehyde （MDA） content in brain tissue by chemical colorimetry. Results （1）Nerve function defects scores：there was no significantly statistical difference between model group and treatment group at 3h after operation （P 〉 0.05 ） , but there were statistical differences at different time points before being killed （ P 〈 0.05 ）. （2） The expression of NGB protein and NGB mRNAN gradually decreased over time. At all time points, the expression of that in model group was more than that in sham group, the expression of that in treatment group was more than that in model group, and the differences were all statistically significant （ P 〈 0.05 ）. （ 3 ） The SOD activity and MDA content gradually decreased over time, At all time points, sham group＇ s SOD activity was more than treatment group＇ s,treatment group＇ s SOD activity was more than model group＇ s, and the differences were all statisti- cally significant （ P 〈 0.05 ）. While at all time points, MDA content in sham group＇ s MDA content was less than that in treatment group＇ s,MDA content in treatment group＇ s MDA content was less than that in model group＇ s, and the differences were all statistically significant （ P 〈 0.05 ）. Conclusion NBP can promote the expression of NGB and ameliorating oxida- tive stress injury.