川芎嗪对大鼠脑缺血再灌注损伤后氧化应激、Ca^(2+)-ATP酶活性及炎症因子的影响

Effect of tetramethylpyrazine on oxidative stress,Ca^(2+)-ATP activity and inflammatory factors after cerebral ischemia/reperfusion injury in rats

目的通过观察脑组织氧自由基、钙离子负荷及炎性反应变化研究川芎嗪对大鼠脑缺血再灌注损伤的治疗作用。方法30只雄性SD大鼠随机分为假手术组、模型组、川芎嗪组(n=10),模型组、川芎嗪组采用大脑中动脉栓塞(MCAO)法建立大鼠脑缺血再灌注模型,24 h后腹腔注射川芎嗪(50 mg/kg,每日1次,连续6 d),造模第1、3、7天进行神经功能评分,造模第7天通过2,3,5-氯化三苯基四氮唑(TTC)染色及扫描脑梗死区光密度计算梗死率,检测脑组织氧自由基一氧化氮(NO)和丙二醛(MDA)水平、钙离子腺苷三磷酸酶(Ca^(2+)-ATP酶)活性、炎症因子基因相对表达水平。结果造模第7天,模型组大鼠神经功能评分、梗死率与假手术组比较均明显升高,差异有统计学意义(P<0.05);与模型组相比,川芎嗪组大鼠神经功能评分和梗死率明显降低,差异有统计学意义(P<0.05),脑组织NO、MDA水平及炎症因子基因表达均下调(P<0.05),超氧化物歧化酶(SOD)和Ca^(2+)-ATP酶活性均升高(P<0.05)。结论川芎嗪通过调节大鼠脑组织氧化和抗氧化平衡,清除氧自由基,提高Ca^(2+)-ATP酶活性,抑制细胞内钙超载,抑制炎性反应等起到改善脑缺血再灌注损伤,保护神经细胞的作用,为探讨临床中药治疗脑卒中机制奠定实验基础。

Objective To investigate the effects of tetramethylpyrazine on treatment in cerebral ischemia/reperfusion injury by the changes of oxygen free radical,calcium overload and inflammatory reaction.Methods Thirty male SD rats were randomly divided into sham operation group,model group and tetramethylpyrazine group(n=10).The model group and tetramethylpyrazine group were established by middle cerebral artery occultation(MCAO).After 24 h,the rats were intraperitoneally injected with ligustrazine(50 mg/kg,once a day,for 6 days).Neurological function score was performed on the first,third and seventh days of modeling.On the seventh day after modeling,the infarction rate was calculated by 2,3,5-triphenyltetrazolium chloride(TTC)staining and scanning optical density of cerebral infarction area.The levels of oxygen free radical nitric oxide(NO)in brain tissue and malondialdehyde(MDA),the activity of calcium adenosine triphosphatase(Ca^(2+)-ATPase)and the relative expression levels of inflammatory factors genes in brain tissues were detected.Results On the 7th day after injury,compared with the sham group,the scores of the nervous function and ratio of cerebral infarct were significantly higher,the difference was statistically significant(P<0.05).The oxidative stress indexes and the expression of inflammatory factors were also significantly changed;compared with the model group,the scores and ratio of treated rats were significantly decreased in the tetramethylpyrazine group,the difference was statistically significant(P<0.05).Also in tetramethylpyrazine group,the levels of NO and MDA as while as the gene expression of inflammatory factors were decreased in the tissues(P<0.05).And the activity of SOD and Ca^(2+)-ATPase were increased compared to the model group(P<0.05).Conclusion Tetramethylpyrazine can improve cerebral ischemia/reperfusion injury and protect nerve cells by regulating the balance of oxidation and antioxidation,scavenging oxygen free radicals,increasing Ca^(2+)-ATPase activity,inhibiting intracellular calcium overload and inhibiting inflammatory reaction,which lay the foundation of study on the mechanism of clinical Chinese medicine in the treatment of stroke.