丙泊酚后处理对大鼠原代皮质神经元氧糖剥夺．复糖复氧后P38蛋白激酶和促凋亡因子Bid、Bim、Puma表达的影响  

Relationships of propofol post-conditioning with expressions of P38 mitogen-activated protein kinase, Bid, Bim and Puma when cortical neurons being exposed to oxygen-glucose deprivation/ resuscitation

作　　者：马育霞[1] 张立民[2] 张冬雪[1] 刘云峰[1] 

机构地区：[1]沧州市中心医院老年内科,061000 [2]沧州市中心医院麻醉科,061000

出　　处：《中华神经医学杂志》2016年第5期439-444,共6页Chinese Journal of Neuromedicine

摘　　要：目的探讨丙泊酚后处理对大鼠原代皮质神经元氧糖剥夺-复糖复氧后P38蛋白激酶（P38MAPK）和促凋亡因子Bid、Bim、Puma表达的影响。 方法体外培养出生24 h内的SD大鼠的皮质神经元并接种于6孔板或96孔板培养皿，分为对照组（C组，正常培养）、氧糖剥夺-复糖复氧组（O组）、氧糖剥夺-复糖复氧＋丙泊酚后处理组（OP组）、氧糖剥夺-复糖复氧＋丙泊酚后处理组＋P38MAPK阻断处理组（OPS组）、P38MAPK阻断处理组（S组）、氧糖剥夺-复糖复氧＋P38MAPK阻断处理组（OS组）。氧糖剥夺-复糖复氧处理是将神经元氧糖剥夺90 min并复糖复氧24 h；丙泊酚后处理为向培养基中加入丙泊酚原液（50 μmol/L）处理2 h；P38MAPK阻断处理为提前1 h向培养皿中加入P38MAPK抑制剂SB202190（50 μmol/L）。收集各组处理好的神经元，采用Annexin V-FITC/PI双染色法测定神经元凋亡率，MTT法测定细胞生存率，乳酸脱氢酶法测定细胞损伤率，JC-1荧光法测定线粒体膜电位水平，荧光素-荧光素酶试剂盒测定ATP含量，Western blotting测定P38MAPK和促凋亡因子Bid、Bim、Puma表达水平。结果与C组比较，O组神经元磷酸化P38MAPK表达升高，凋亡率升高，存活率降低，损伤率升高，线粒体膜电位水平和ATP含量降低，促凋亡因子Bid、Bim、Puma表达水平增加，差异均有统计学意义（P〈0.05）。与O组比较，OP组神经元磷酸化P38MAPK表达升高，凋亡率降低，存活率升高，损伤率降低，线粒体膜电位水平和ATP含量升高，促凋亡因子Bid、Bim、Puma表达水平降低，差异均有统计学意义（P〈 0.05）。与OP组比较，OPS组神经元凋亡率升高，存活率降低，损伤率升高，线粒体膜电位水平和ATP含量降低，促凋亡因子Bid、Bim、Puma表达水平增加，差异均有统计学意义（P〈0.05）。与O组比较，OS组神经元凋亡率升高，存活率降低，损伤率升高，线Objective To explore the relations ofpropofol post-conditioning with expressions of P38 mitogen-activated protein kinase （P38MAPK）, Bid, Bim and Puma when cortical neurons being exposed to oxygen-glucose deprivation/resuscitation （OGD）. Methods Cortical neurons from SD rats within 24 h of birth were cultured in vitro and inoculated at 6- or 96- well plates; and then, they were divided into control （C） group, OGD （O） group, OGD＋propofolpost-conditioning （OP） group, OGD＋propofol post-conditioning＋SB202190 （OPS） group, SB202190 treatment （S） group and OGD＋SB202190 （OS） group. In vitro cultured OGD cortical neurons for 90 min and resuscitaion for 24 h were established in the above groups excepted for C group; after OGD exposure, the cells in OP and OPS groups were treated with 50 μmol/L propofol for 2 h; the cells in OPS, S and OS groups were treated with SB202190 （50 μmol/L） to inhibit phosphorylation of P38MAPK. The neurons in the above groups then were collected. Neuronal apoptosis, viability and injury were assessed by Annexin V-FITC/PI asaay, MTT assay and lactate dehydrogenase （LDH） release assay, respectively. Mitochondrial membrane potential （MMP） was assessed by JC-1 fluorescece assay. ATP content was assessed by fluorescein-luciferase assay kit. The protein expressions of P38MAPK, Bid, Bim and Puma were assessed by Western blotting. Results As compared with those in the C group, significant increase of neuronal apoptosis, injury and expressions of Bid, Bim, Puma and phosphorylation of P38MAPK but decrease of viability, MMP and ATP content were noted in O group （P〈0.05）. As compared with those in the O group, significant decrease of neuronal apoptosis, injury and expressions of Bid, Bim, Puma, but increase of viability, MMP, ATP content and phosphorylation of P38MAPK were noted in the OP group （P〈0.05）. As compared with those in the OP group, significant increase of neuronal apoptosis, injury and expressions of Bid, Bim, Puma, but decrease of

关 键 词：丙泊酚 氧糖剥夺.复糖复氧 P38蛋白激酶 BID BIM Puma 细胞凋亡 

分 类 号：R96[医药卫生—药理学]