吡格列酮对脑缺血再灌注大鼠的影响研究

Effect of pyglidone on cerebral ischemia reperfusion rats

目的研究吡格列酮对脑缺血再灌注大鼠神经功能、神经元凋亡及凋亡相关蛋白表达的影响。方法45只清洁级健康雄性SD大鼠以Longa线栓法构建脑缺血再灌注损伤模型,42只建模成功并随机分为模型组和低、高剂量实验组,各14只。低、高剂量实验组分别于术前1,2,3 d及术前1 h灌胃10,15mg·kg^-1吡格列酮,每天1次。另选10只雄性SD大鼠仅分离血管,但不结扎阻塞大脑中动脉血供,作为假手术组,于同时间点灌胃给予等量生理盐水。于缺血再灌注22 h后以Zea-Longa评分法评估大鼠神经功能;以DNA断裂的原位末端标记(TUNEL)法检测大鼠缺血侧脑组织神经元凋亡;以免疫组化法检测大鼠缺血侧脑组织凋亡相关蛋白表达。结果再灌注22 h时,假手术组、模型组及低、高剂量实验组Zea Longa评分分别为(0.93±0.15),(3.46±0.41),(3.01±0.33),(2.24±0.27)分;神经元凋亡指数(AI)分别为2.03±0.96,69.87±10.98,32.41±9.65,20.05±4.35;缺血侧脑组织B淋巴细胞瘤-2(Bcl-2)蛋白阳性细胞比例分别为(0.45±0.04)%,(0.18±0.01)%,(0.23±0.02)%,(0.35±0.03)%;Bcl-2相关蛋白X(Bax)蛋白阳性细胞比例分别为(0.11±0.01)%,(0.36±0.02)%,(0.13±0.01)%,(0.09±0.01)%。假手术组和低、高剂量实验组与模型组比较,差异均有统计学意义,且高、低剂量实验组相比差异均有统计学意义(均P<0.05)。结论高剂量吡格列酮可有效上调脑组织Bcl-2表达,下调Bax表达,抑制神经元凋亡,改善脑缺血再灌注损伤大鼠神经功能。

Objective To explore the effect of pyglidone on the nerve function,neuronal apoptosis and apoptosis related proteins expression of cerebral ischemia reperfusion rats.Methods Forty-five clean level healthy male SD rats were built cerebral ischemia reperfusion model by Longa suture method,the 42 successfully modeling rats were divided randomly into model group and low,high dose experimental groups,14 rats in each group.Low,high dose experimental groups were intragastricly administrated 10,15 mg·kg^-1 pyglidone respectively at 1,2,3 d and 1 h pre-operation,1 time·d^-1.Another 10 male SD rats were acted as sham-operation with only separated blood vessels,but didn’t ligated and blocked the artery of the brain and intragastricly administrated equivalent normal saline at the same time point.The nerve function was evaluated by Zea-Longa scoring 22 h after cerebral ischemia reperfusion;the ischemic lateral brain tissue nerve cell apoptosis was detectedby terminal dexynucleotidyl transferase(Td T)-mediated d UTP nick end labeling(TUNEL);the apoptosis related proteins expression was detected by immunohistochemistry.Results At 22 h after cerebral ischemia reperfusion,the Zea Longa scores in sham-operation group,model group and low,high dose experimental groups were(0.93±0.15),(3.46±0.41),(3.01±0.33),(2.24±0.27)scores;neuron apoptosis indexes(AI)were 2.03±0.96,69.87±10.98,32.41±9.65,20.05±4.35;the ischemic lateral brain tissue B cell lymphoma-2 gene(Bcl-2)protein positive cell proportion were(0.45±0.04)%,(0.18±0.01)%,(0.23±0.02)%,(0.35±0.03)%;Bcl-2 associated X protein(Bax)protein positive cell proportion were(0.11±0.01)%,(0.36±0.02)%,(0.13±0.01)%,(0.09±0.01)%.The difference between sham-operation group and model and low,high dose experimental groups were all significant,and the difference between low and high dose experimental groups were all significant(all P<0.05).Conclusion High dose pyglidone can up-regulate brain tissue Bcl-2 expression effectively,down-regulate Bax expression,inhibit neuronal apoptosis,and then improve the nerve function of cerebral ischemia reperfusion rats.