SIRT1-ERK1∕2通路与竹节参皂苷减轻异氟醚致胎鼠海马神经元毒性的关系:离体实验  

Relationship between SIRT1-ERK1/2pathway and chikusetsu saponin IVa-induced reduction of isoflurane-elicited neurotoxicity in fetal rats:an in vitro experiment

作　　者：方茜 高杰 李世勇[1] 张杰 杨春 罗爱林[1] Fang Xi;Gao Jie;Li Shiyong;Zhang Jie;Yang Chun;Luo Ailin(Department of Anesthesiology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉科,武汉市430030

出　　处：《中华麻醉学杂志》2018年第8期908-910,共3页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81571047,81400882,81500931).

摘　　要：目的 采用离体实验评价沉默信息调节因子1(SIRT1)-细胞外信号调节激酶1∕2(ERK1∕2)通路与竹节参皂苷减轻异氟醚致胎鼠海马神经元毒性的关系.方法 孕16~18 d大鼠,剖腹取出胎鼠,取海马神经元,原代培养7 d,采用随机数字表法分为3组(n=6):对照组(Con组)、异氟醚组(Iso组)和竹节参皂苷+异氟醚组(ChIV+Iso组).Con组常规培养6 h;Iso组将培养基放入充有1.8%异氟醚的培养箱中孵育6 h;ChIV+Iso组培养基中加入竹节参皂苷25μg∕ml孵育6 h,然后将其放入充有1.8%异氟醚的培养箱中孵育6 h.收集神经元培养液上清,测定LDH释放量,采用CCK-8法测定神经元活力,采用Western blot法测定神经元SIRT1、ERK1∕2和磷酸化ERK1∕2(p-ERK1∕2)的表达水平.结果 与Con组比较,Iso组神经元活力降低,LDH释放量升高,SIRT1和p-ERK1∕2的表达下调(P<0.05);与Iso组比较,ChIV+Iso组神经元活力升高,LDH释放量降低,SIRT1和p-ERK1∕2的表达上调(P<0.05).结论 竹节参皂苷减轻异氟醚致胎鼠海马神经元毒性的机制可能与激活SIRT1-ERK1∕2通路有关.Objective To evaluate the relationship between silent information regulator 1 ( SIRT1)-extracellular-regulated kinase1∕2 ( ERK1∕2) pathway and chikusetsu saponin IVa-induced reduc-tion of isoflurane-elicited neurotoxicity in fetal rats in an in vitro experiment. Methods The hippocampal neurons isolated from rats at 16-18 days of gestation were primarily cultured for 7 days and divided into 3 groups ( n = 6 each) using a random number table method: control group ( Con group) , isoflurane group (Iso group) and chikusetsu saponin IVa plus isoflurane group (ChIV+Iso group). Hippocampal neurons were cultured routinely for 6 h in Con group. Hippocampal neurons were exposed to 1. 8% isoflurane for 6 h in an incubator in Iso group. Chikusetsu saponin IVa 25μg∕ml was added to the culture medium, and hipp-ocampal neurons were incubated for 6 h and then exposed to 1. 8% isoflurane for 6 h in an incubator in ChIV+Iso group. The supernatant was collected for determination of the amount of lactic dehydrogenase ( LDH) released, neuronal viability ( by CCK-8) and expression of SIRT1, ERK1∕2 and phosphorylated ERK1∕2 ( p-ERK1∕2) ( by Western blot) . Results Compared with Con group, the neuronal viability was significantly decreased, the amount of LDH released was increased, and the expression of SIRT1 and p-ERK1∕2 was down-regulated in Iso group ( P<0. 05) . Compared with Iso group, the neuronal viability was significantly increased, the amount of LDH released was decreased, and the expression of SIRT1 and p-ERK1∕2 was up-regulated in ChIV+Iso group ( P<0. 05) . Conclusion The mechanism by which chikuset-su saponin IVa reduces isoflurane-elicited neurotoxicity is related to activating SIRT1-ERK1∕2 pathway in fe-tal rats in an in vitro experiment.

关 键 词：异氟醚 抗衰老酶 细胞外信号调节MAP激酶类 药物毒性 神经元 

分 类 号：R614[医药卫生—麻醉学]