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目的观察金钗石斛生物总碱(DNLA)预防性给药对大鼠大脑中动脉阻塞的保护作用及相关机制。方法 104只SD大鼠随机分成假手术组,模型组,DNLA 20、40和80 mg·kg-1组。每日2次灌胃给药2 d后,线栓法建立大鼠右侧大脑中动脉阻塞的局灶性脑缺血模型。24 h后观察大鼠神经功能症状,HE染色观察组织病理变化,2,3,5-三苯基氯化四氮唑(TTC)染色测量脑梗死体积,测定MDA、NO的含量和SOD、NOS的活性,并分析MDA与SOD的比例变化,PCR法分析脑组织中caspase-3和caspase-8的mRNA表达。结果 DNLA 80 mg·kg-1组神经功能症状评分下降,组织病理损伤明显减轻;DNLA 40 mg·kg-1组和80 mg·kg-1组脑梗死体积缩小,MDA、NO的含量减少,NOS的活性降低,SOD的活性升高,MDA与SOD的比值减少,caspase-3、caspase-8 mRNA的表达下调,尤其40 mg·kg-1和80 mg·kg-1剂量组更为明显。结论 DNLA预防性给药对大鼠急性脑缺血有保护作用,其机制与抗氧化应激、清除自由基及降低大鼠脑内caspase-3、caspase-8 mRNA表达有关。
Objective To observe the protective effect of DNLA on the middle cerebral artery occlusion in rats and related mechanisms. Methods A total of 104 SD rats were randomly divided into sham-operated group, model group, DNLA 20, 40 and 80 mg·kg-1 groups. After 2 days of intragastric administration twice a day, a model of focal cerebral ischemia in the right middle cerebral artery was established by a suppository method. After 24 hours, the neurological symptoms of rats were observed. Histopathological changes were observed by HE staining. Infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. The contents of MDA and NO and SOD, NOS were measured. The activity and the ratio of MDA to SOD were analyzed. The mRNA expression of caspase-3 and caspase-8 in brain tissue was analyzed by PCR. Results The neurological symptom scores of DNLA 80 mg·kg-1 group decreased, and the histopathological injury was significantly reduced. The infarct volume was reduced and the contents of MDA and NO were decreased in DNLA 40 mg·kg-1 and 80 mg·kg-1 groups. The activity decreased, the activity of SOD increased, the ratio of MDA to SOD decreased, the expression of caspase-3 and caspase-8 mRNA was down-regulated, especially 40 mg·kg-1 and 80 mg·kg-1. Conclusion Prophylactic administration of DNLA has a protective effect on acute cerebral ischemia in rats, and its mechanism is related to antioxidative stress, scavenging free radicals and reducing the expression of caspase-3 and caspase-8 mRNA in rat brain.