依达拉奉预处理离体乳鼠脑皮质细胞对脑缺血再灌注损伤的保护作用

A Study on the Protection of Edaravone Pretreatment against Cerebral Ischemia-reperfusion Injury

目的:探讨不同浓度依达拉奉预处理离体乳鼠脑皮质细胞对脑缺血-再灌注损伤的保护作用。方法:取出生24h以内的新生SD乳鼠的脑皮质细胞,体外培养至第7天,随机分为5组:A组(正常对照组);B组(药物损伤组);C1组(50μmol/L依达拉奉预处理组);C2组(100μmol/L依达拉奉预处理组);C3组(200μmol/L依达拉奉预处理组)。C1、C2、C3组第7天用依达拉奉预处理,24h后B、C1、C2、C3组予200μmol/L谷氨酸损伤0.5h,所有组换正常培养液继续培养24h后观察神经细胞存活率(MTT法)、乳酸脱氢酶(LDH)漏出率、细胞凋亡率和HE染色后倒置相差显微镜下细胞形态变化。结果:依达拉奉预处理各组MTT含量不同程度高于损伤组,LDH漏出量和凋亡细胞百分比不同程度低于损伤组,倒置相差显微镜下见预处理组细胞形态受损较药物损伤组轻,3个预处理组以C2、C3组效果明显,但两组差别无统计学意义。结论:采用所测浓度的依达拉奉提前24h预处理离体幼鼠脑皮质细胞,对脑缺血-再灌注损伤有明显的保护作用。

Objective： To study the protective effect of edaravone pretreatment against cerebral ische-mia-reperfusion caused injury in.cultured cortical ceils of suckling rat brain. Methods： Cortical cells of SD rat within 24 hours after birth were cultured for 7 days, and then were randomly divided into 5 groups： group A （control group）, group B （ischemia-reperfusion group）, groups C1, C2, and C3 （edaravone pretreatment groups, 50umol/L for group CI, 100umol/L for group C2, 200 μmol/L for group C3 ）. After cells in groups C1 , C2, and C3 were treated by corresponding dosages of edaravone for 24 hours, glutamate （200 p.mol/L） was added into culturing media of groups B, C1, C2, and Ca. In half an hour later, culture media of all groups were substituted by fresh normal medium. After an-other 24 hours, cortical cell survival rates, LDH efflux rates and cell apoptosis rates were determined, and the cell morphous was observed with the help of HE staining. Results： The survival rates were higher, and the LDH efflux rates and apoptosis rates were lower in edaravone groups than those of group B; The cell destruction in edaravone treated groups was lighter than that in group B. The protective results of groups C2 and C3 was obvious, and there was no significant difference between the 2 groups. Conclusions： The pretreatment of edaravone with the tested concentrations in 24 hours before has obvious protective effect to cultured cortical cells against the cerebral ischemia-reperfusion caused injury ;