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目的:观察参芎注射液对大鼠局灶性脑缺血再灌注后神经细胞凋亡及内质网应激相关因子葡萄糖调节蛋白/免疫球蛋白结合蛋白(GRP78/Bip)表达的影响。方法:100只雄性SD大鼠随机分为正常组、假手术组、脑缺血再灌注组、参芎治疗组;后两组根据再灌注时间不同各分为6、12、24、72 h四个亚组;采用大鼠大脑中动脉线栓法制备局灶性脑缺血再灌注模型。TUNEL法观察细胞凋亡情况;免疫组化和RT-PCR法检测各实验组中缺血周围区GRP78/Bip的表达。结果:TUNEL法表明参芎治疗组大鼠大脑神经细胞凋亡程度较缺血再灌注组明显减轻。免疫组化和RT-PCR检测均发现各时间点缺血再灌注组大鼠GRP78/Bip表达高于假手术组及正常组;脑缺血再灌注组及参芎治疗组GRP78/Bip的表达于缺血后12 h最高,72 h恢复至正常水平,且均呈现先升高后降低的趋势;各时间点缺血再灌注组GRP78/Bip表达均高于参芎治疗组。结论:再灌注损伤后12 h内出现GRP78/Bip表达升高,参芎注射液可以下调其表达,从而可能通过减轻内质网应激而减轻缺血再灌注损伤起到神经元保护的作用。
Objective: To observe the effect of Shenqi injection on neuronal apoptosis and the expression of endoplasmic reticulum stress-related factor, glucose regulatory protein/immunoglobulin-binding protein (GRP78/Bip) after focal cerebral ischemia-reperfusion in rats. Methods: 100 male SD rats were randomly divided into normal group, sham operation group, cerebral ischemia-reperfusion group and Shenqi treatment group. The latter two groups were divided into 6, 12, 24, and 72 hours according to different reperfusion time. A subgroup; Focal cerebral ischemic reperfusion model was established by the middle cerebral artery occlusion method. Apoptosis was observed by TUNEL method. The expression of GRP78/Bip in the peripheral ischemia region was detected by immunohistochemistry and RT-PCR. RESULTS: TUNEL assay showed that the degree of cerebral neuron apoptosis in the Shenqi treatment group was significantly less than that in the ischemia-reperfusion group. Immunohistochemistry and RT-PCR showed that the expression of GRP78/Bip in ischemia-reperfusion rats was higher than that in sham-operation group and normal group at each time point; the expression of GRP78/Bip in cerebral ischemia-reperfusion group and Shenqi treatment group was significantly increased. The highest level was at 12 h after ischemia, and it returned to normal at 72 h, and both showed a trend of increasing first and then decreasing. The expression of GRP78/Bip in ischemia-reperfusion group was higher than that in Shenqi treatment group at each time point. Conclusion: The expression of GRP78/Bip increased within 12 h after reperfusion injury. Shenqi injection can down-regulate the expression of GRP78/Bip, which may reduce the ER stress and reduce the ischemia-reperfusion injury and play a role in neuroprotection.