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目的研究羟基红花黄色素A(HSYA)与芍药苷联合用药对急性局灶性脑缺血再灌注(I/R)损伤大鼠的协同保护作用,并探讨其作用机制。方法 110只雄性SD大鼠,随机分为6组:HSYA组(5.0 mg/kg)、芍药苷组(5.0 mg/kg)、HSYA与芍药苷联合用药组(合用组,各5.0 mg/kg)、银杏内酯组(5 mg/kg)、模型组和假手术组。采用改良线栓法复制大鼠大脑中动脉栓塞(MCAO)再灌注模型。脑缺血1 h再灌注6 h后进行神经功能缺失评分及尾静脉注射给药;给药7 d后进行神经功能缺失评分;氯化三苯基四氮唑(TTC)染色法检测脑梗死面积;免疫组化(IHC)法检测磷酸化Akt1蛋白的表达量的变化情况。结果与假手术组比较,模型组大鼠治疗前评分明显高于假手术组,差异具有统计学意义(P<0.01),说明造模成功;与模型组比较,银杏内酯组、合用组、芍药苷组和HSYA组能不同程度地抑制大鼠体质量下降(P<0.05,P<0.01),改善脑缺血再灌注大鼠的神经功能缺失症状(P<0.05,P<0.01),降低皮质区神经细胞的损伤程度,减小脑梗死面积(P<0.05,P<0.01),促进磷酸化Akt1蛋白的表达(P<0.01);HSYA组、芍药苷组、银杏内酯组磷酸化Akt1蛋白的表达较合用组低(P<0.01)。结论 HSYA与芍药苷联合用药对脑缺血再灌注大鼠的脑损伤具有一定的协同保护作用,二者联合用药的作用优于单独用药,可能与二者联合用药发挥活血化瘀与神经保护作用有关。
Objective To study the synergistic protective effect of hydroxysafflor yellow A (HSYA) and paeoniflorin on acute focal cerebral ischemia-reperfusion (I / R) injury in rats and its mechanism. Methods 110 male SD rats were randomly divided into 6 groups: HSYA group (5.0 mg / kg), paeoniflorin group (5.0 mg / kg), HSYA combined with paeoniflorin group , Ginkgolide group (5 mg / kg), model group and sham operation group. Rat model of middle cerebral artery occlusion (MCAO) reperfusion was reproduced by modified thread method. Neurological deficit scores and caudal vein injection were administered 6 h after reperfusion for 1 h after cerebral ischemia. Neurological deficit score was measured after 7 d administration. TTC staining was used to detect infarct size The expression of phosphorylated Akt1 protein was detected by immunohistochemistry (IHC). Results Compared with the sham operation group, the scores of the model group before treatment were significantly higher than those of the sham operation group (P <0.01), indicating that the model was successful. Compared with the model group, the ginkgolide group, combined group, Paeoniflorin and HSYA could inhibit the body weight of rats to some extent (P <0.05, P <0.01), and improve the symptoms of neurological deficits (P <0.05, P <0.01) and decrease in rats with cerebral ischemia-reperfusion (P <0.05, P <0.01), and promoted the phosphorylation of Akt1 protein (P <0.01). The phosphorylation of Akt1 in HSYA group, paeoniflorin group and ginkgolide group The protein expression was lower than that of the combined group (P <0.01). Conclusion Combined administration of HSYA and paeoniflorin has a synergistic protective effect on brain injury induced by cerebral ischemia and reperfusion in rats. The combination of HSYA and paeoniflorin has a better effect than single administration on the brain injury, and may be used in combination with the two drugs to promote blood circulation and neuroprotection related.