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目的观察复方蒲黄颗粒对大鼠脑缺血再灌注后热休后蛋白70(HSP-70)及其mRNA表达的影响。方法75只雄性SD大鼠,采用大脑中动脉线栓法建立局灶性脑缺血再灌注大鼠模型,随机分为假手术组、模型组,蒲黄颗粒大、中、小剂量组,每组15只。各组大鼠分别在缺血2小时再灌注6、24、72小时断头取脑组织,每时间点5只。再灌注24小时进行神经功能缺陷评分,采用Nissl染色观察神经元形态学变化;采用原位杂交检测HSP-70 mRNA的表达;采用免疫组化法检测HSP-70蛋白的表达。结果蒲黄颗粒大、中剂量组在缺血再灌注24小时时神经功能改善显著优于模型组(P<0.05);Nissl染色结果显示,模型组大脑皮层神经元形态不规则,体积缩小,胞核固缩深染,细胞间质水肿,尼氏体减少,可见明显的神经元丢失,而蒲黄颗粒各剂量组未见明显的神经元损伤和丢失;模型组在再灌注6～72小时HSP-70及其mRNA的表达均增加,与假手术组比较各时间点差异均具有统计学意义(P<0.01),蒲黄颗粒各剂量均能增强HSP-70及其mRNA的表达,与模型组各时间点的差异均具有统计学意义(P<0.05)。结论复方蒲黄颗粒能促进脑缺血再灌注后HSP-70的表达,减轻缺血再灌注损伤。
Objective To observe the effect of Fufang Puhuang Granule on the expression of HSP-70 and its mRNA after heat shock after cerebral ischemia-reperfusion in rats. Methods Seventy-five male Sprague-Dawley rats were established by middle cerebral artery occlusion to establish a model of focal cerebral ischemia and reperfusion. They were randomly divided into sham-operated group and model group. Puhuang granules were divided into large, medium and small dose groups. Group 15 only. Rats in each group were decapitated at 6, 24, and 72 hours after ischemia for 2 hours and 5 rats were sacrificed at each time point. Neurological deficit scores were performed 24 hours after reperfusion. Nissl staining was used to observe neuronal morphological changes. In situ hybridization was used to detect the expression of HSP-70 mRNA. Immunohistochemistry was used to detect the expression of HSP-70 protein. Results Compared with the model group, the neurological function of Puhuanghuangda group and Dazhong group improved significantly at 24 hours after ischemia and reperfusion (P<0.05). Nissl staining results showed that the cerebral cortex neurons in the model group were irregular in shape and the cell size was reduced. Deep staining of nuclear condensation, interstitial edema, decreased Nissl body, visible neuron loss, and no obvious neuronal damage and loss in each dose group of Puhuang granule; model group in the reperfusion 6-72 hours HSP The expression of -70 and its mRNA increased, and there were significant differences between the time points (P<0.01) and the sham operation group. Each dose of Puhuang granules enhanced the expression of HSP-70 and its mRNA, and the model group. The differences at each time point were statistically significant (P<0.05). Conclusion Compound Puhuang granules can promote the expression of HSP-70 and reduce the ischemia-reperfusion injury after cerebral ischemia-reperfusion.