摘要
目的观察羟基红花黄色素A(HYSA)腹腔注射对大鼠脑缺血再灌注损伤的预防作用,并探讨其机制。方法60只SD大鼠随机分成假手术组、模型组、丁苯酞组、低剂量HYSA组、高剂量HYSA组,每组12只。假手术组、模型组腹腔注射0.9%NaCl溶液,丁苯酞组腹腔注射丁苯酞软胶囊混悬液,低剂量HYSA组腹腔注射5.0 mg/mL的HYSA,高剂量HYSA组腹腔注射10.0 mg/mL的HYSA,每天1次,连续5 d后,模型组、丁苯酞组、低剂量HYSA组、高剂量HYSA组使用小修正方法进行脑缺血再灌注处理,假手术组不处理。各组大鼠处理完毕后清醒4 h,采用Longa评分法对各组大鼠进行神经功能评分。处理完毕后,采用HE染色法观察大鼠脑组织形态学变化,采用RTqPCR法检测大鼠脑组织中Bax、Bcl-2、Caspase-3、c-fos mRNA,采用Western Blotting法检测大鼠脑组织中Bax、Bcl-2、Caspase-3、c-fos蛋白。结果模型组大鼠神经功能评分显著高于其余各组(P均<0.05),高剂量HYSA组、丁苯酞组大鼠神经功能评分显著低于低剂量HYSA组(P均<0.05)。模型组大鼠脑组织细胞间隙明显增大,细胞核减少且深染,细胞形态不规则;低剂量HYSA组较模型组细胞形态有一定改善,但部分细胞核仍浓缩深染;高剂量HYSA组和丁苯酞组细胞坏死明显改善,细胞形态恢复正常,细胞排列整齐。模型组大鼠脑组织中Bax、Caspase-3、c-fos mRNA和蛋白的相对表达量显著高于其余各组(P均<0.05),Bcl-2 mRNA和蛋白的相对表达量显著低于其余各组(P均<0.05);高剂量HYSA组、丁苯酞组大鼠脑组织中Bax、Caspase-3、c-fos mRNA和蛋白的相对表达量显著低于低剂量HYSA组(P均<0.05),Bcl-2 mRNA和蛋白的相对表达量显著高于低剂量HYSA组(P均<0.05)。结论高剂量(10.0 mg/mL)HYSA腹腔注射对大鼠脑缺血再灌注损伤有预防作用,与丁苯酞效果相当;HYSA预防大鼠脑缺血再灌注损伤的作用机制可能与调节脑组织中Bax、Bcl-2、Caspase-3、c-fos的表达有关。
Objective To observe the preventive effect of intraperitoneal injection of hydroxy safflower yellow A(HYSA) on cerebral ischemia-reperfusion injury in rats and to explore its mechanism.Methods Sixty SD rats were randomly divided into the sham operation group,model group,butylphthalide group,low-dose HYSA group,and high-dose HYSA group,with 12 rats in each group.The rats in the sham operation group and model group were intraperitoneally injected with 0.9% NaCl solution,the rats in the butylphthalide group were intraperitoneally injected with butylphthalide soft capsule suspension,the rats in the low-dose HYSA group were intraperitoneally injected with 5.0 mg/mL HYSA,and the rats in the high-dose HYSA group were intraperitoneally injected with 10.0 mg/mL HYSA,once a day.After five consecutive days,the rats in the model group,butylphthalide group,low-dose HYSA group and high-dose HYSA group were treated with small correction method for cerebral ischemia-reperfusion,and the rats in the sham operation group were not treated.Rats in each group were awake for 4 h after treatment,and the neurological function of rats in each group was scored by Longa score method.After treatment,the morphological changes of rat brains were observed by HE staining;Bax,Bcl-2,Caspase-3 and c-fos mRNA were detected by RT-qPCR;and Bax,Bcl-2,Caspase-3 and c-fos proteins were detected by Western blotting.Results The neurological function score of the model group was significantly higher than that of the other groups(all P<0.05),and those of the high-dose HYSA group and butylphthalide group were significantly lower than that of the low-dose HYSA group(all P<0.05).In the model group,the intercellular space of brain tissues increased significantly,the nucleus decreased and deeply stained,and the cell morphology was irregular.The cell morphology of the low-dose HYSA group was better than that of the model group,but some nuclei were still concentrated and deeply stained.In the high-dose HYSA group and butylphthalide group,cell necrosis was significantly improved,cell morphology returned to normal,and cells were arranged neatly.The relative expression levels of Bax,Caspase-3,c-fos mRNA and protein in the model group were significantly higher than those in the other groups(all P<0.05),and the relative expression levels of Bcl-2 mRNA and protein were significantly lower than those in the other groups(all P<0.05).The relative expression levels of Bax,Caspase-3,and c-fos mRNA and protein in the high-dose HYSA group and butylphthalide group were significantly lower than those in the low-dose HYSA group(all P<0.05),and the relative expression levels of Bcl-2 mRNA and protein were significantly higher than those in the low-dose HYSA group(all P<0.05).Conclusion Intraperitoneal injection of high-dose HYSA(10.0 mg/mL) has a preventive effect on cerebral ischemia-reperfusion injury in rats,which is equivalent to that of butylphthalide;the mechanism of HYSA for preventing cerebral ischemia-reperfusion injury in rats may be related to regulating the expression levels of Bax,Bcl-2,Caspase-3 and c-fos in the brain tissues.
作者
王飞
乔栎
刘建雄
张宏星
WANG Fei;QIAO Li;LIU Jianxiong;ZHANG Hongxing(The First Clinical Medical College of Gansu University of Traditional Chinese Medicine,Lanzhou 730030,China;不详)
出处
《山东医药》
CAS
2022年第9期47-51,共5页
Shandong Medical Journal
基金
甘肃省人民医院院内科研基金项目(20GSSY2-2)。
关键词
羟基红花黄色素A
脑缺血再灌注损伤
缺血性脑卒中
脑神经
Hydroxy safflower yellow A
cerebral ischemia-reperfusion injury
ischemic stroke
cerebral nerve