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作 者:荣妍[1] 何昱[1] 程兰[1] 王慧君 虞立[1] 杨洁红 周惠芬[1] 万海同[1]
机构地区:[1]浙江中医药大学,杭州310053
出 处:《中国药学杂志》2018年第3期199-204,共6页Chinese Pharmaceutical Journal
基 金:国家自然科学基金资助项目(81374053;81630105);浙江省自然科学基金资助项目(LZ17H270001);浙江省卫生高层次创新人才培养工程资助项目(2015年度)
摘 要:目的探究养阴通脑颗粒主要有效成分葛根素、川芎嗪、黄芪甲苷、梓醇的正交配伍组合对SD大鼠脑缺血再灌注损伤的保护作用机制,并选出较优组合。方法将SD大鼠随机分为假手术组、模型组、9个不同比例正交配伍组和阳性对照组。建立大鼠大脑中动脉局灶性栓塞(MCAO)模型,以L_9(3~4)正交表设计安排成分配伍给药,动物清醒后观察大鼠神经功能缺损症状;3d后观察海马区脑组织的病理变化;TTC染色检测脑梗死体积;免疫组织化学法来评估缺血侧脑组织NLRP3蛋白表达,RT-PCR法检测海马区脑组织ASC、caspase-1、IL-1β、IL-18、MMP-9 mRNA表达。采用极差分析法分析正交试验结果。结果正交配伍各组能有效改善脑缺血再灌注损伤的神经功能缺失,降低脑梗死体积,抑制NLRP3炎性体的表达,降低ASC、caspase-1、IL-1β、IL-18、MMP-9基因的表达。正交试验结果分析显示,其较优组合为川芎嗪100 mg·kg^(-1)、葛根素20 mg·kg^(-1)、黄芪甲苷80 mg·kg^(-1)、梓醇20 mg·kg^(-1)。结论养阴通脑颗粒主要有效成分配伍对脑缺血再灌注损伤大鼠有明显的保护作用,其作用机制可能与抑制NLRP3炎性体,下调ASC、caspase-1、IL-1β、IL-18、MMP-9的表达有关。OBJECTIVE To explore the protection mechanism of orthogonal compatibility of puerarin(A) ,ligustrazine(B), astragaloside IV (C) and catalpolon( D), the main active ingredients from yangyin tongnao granules, on SD rats damaged by ischemie reperfusion injury, and select the optimal combination. METHODS SD rats were randomly divided into sham operation group, model group, orthogonal compatibility group and positive control group. The rat middle cerebral artery occlusion (MCAO) model was established. The compatibility of components was arranged by L9 (34) orthogonal design. The symptoms of neurological deficit in rats and the pathological changes in hippocampus were observed; TTC staining was used to detect the cerebral infarction volume. IHC was used to evaluate the expression of NLRP3 protein ischemie brain tissue. RT-PCR was used to detect the expressions of ASC, caspase-1, IL- 1, mRNA, IL-1β and MMP-9 in the hippocampal of brain tissue. The results of orthogonal test were analyzed by range analysis. RESULTS The orthogonal combination groups could effectively improve neurological deficits in cerebral ischemia reperfusion injury, reduce infarction volume, inhibit the expression of NLRP3 inflammasome, decrease the expression of ASC, caspase-1, IL-1, IL-18 and MMP-9. The analysis of test results showed that the combination with ligustrazine 100 mg · kg-1 ,puerarin 20 mg · kg-1 , astragaloside IV 80 mg · kg - 1 and catalpol 20 mg · kg- 1 was the superior one. CONCLUSION The main active ingredients combination of yangyin tongnao granules played a protective mechanism on focal cerebral ischemia reperfusion injury rats. Its mechanism might be related to inhibition of NLRP3 inflammasome,down-regulation of ASC, easpase-1, IL-1, IL-18 and MMP-9.
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