国际麻醉学与复苏杂志   2019, Issue (7): 0-0
    
丙泊酚通过细胞外信号调节激酶信号通路上调海马星形胶质细胞脑源性神经营养因子表达和分泌
顾新宇, 朱浩, 杨立群, 俞卫锋1()
1.上海交通大学医学院附属仁济医院
Propofol upregulates the expressions and secretions of brain derived neurotrophic factor in rat hippocampal astrocytes via extracellular signal-regulated kinase signaling pathway
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摘要：

目的　检测丙泊酚对大鼠海马及其星形胶质细胞脑源性神经营养因子（brain derived neurotrophic factor, BDNF）表达分泌的影响，并探讨其中涉及的信号分子。　方法　按随机数字表法将24只SD大鼠分为3组（每组8只），根据丙泊酚给予后不同时间检测点分别命名为：注射丙泊酚后即刻检测组（0 min组）、注射丙泊酚后45 min检测组（45 min组）、注射丙泊酚后90 min检测组（90 min组）。将丙泊酚注射液（10 g/L）以100 mg/kg腹腔注射后，用定量PCR检测大鼠海马BDNF的mRNA水平，用免疫组织化学法检测胶质纤维酸性蛋白（glial fibrillary acidic protein, GFAP）表达。体外实验中培养的细胞分为3组：对照组（Ctrl组）、丙泊酚组（Pro组）和抑制剂PD98059+丙泊酚组（Pro-PD组）。Pro-PD组先用10 μmol/L 细胞外信号调节激酶（extracellular signal-regulated kinase, ERK）抑制剂PD98059预处理培养的原代大鼠海马星形胶质细胞2 h，随后用10 μmol/L丙泊酚继续孵育24 h。检测各组细胞凋亡及BDNF分泌情况。　结果　与0 min 组比较，45 min 组、90 min 组大鼠海马BDNF mRNA 水平分别为1.20±0.05和0.86±0.04（P<0.05），且BDNF mRNA 水平均呈时间依赖性变化，先增高，而后降低。免疫组织化学法检测显示45 min 组GFAP 表达升高。体外实验显示，Ctrl组细胞存活率为（91.2±2.4）%，Pro组细胞存活率为（90.3±3.0）%，两组差异无统计学意义（P>0.05）。与Ctrl组比较，Pro组海马星形胶质细胞的BDNF mRNA水平为1.15±0.04（P<0.05），而Pro-PD组细胞BDNF的mRNA水平为0.92±0.05（P＞0.05），且显著低于Pro组（P＜0.05）。Pro组海马星形胶质细胞BDNF的分泌水平为（62±4） ng/L，高于Ctrl组（49±3） ng/L（P<0.05）；而Pro-PD组细胞BDNF的浓度为（44±3） ng/L，与Ctrl组差异无统计学意义（P>0.05），但显著低于Pro组（P＜0.05）。　结论　丙泊酚激活海马星形胶质细胞，并通过ERK信号通路提高海马星形胶质细胞BDNF的表达和分泌。
关键词： 丙泊酚；　星形胶质细胞；　细胞外信号调节激酶；　脑源性神经营养因子
Abstract:

Ｏｂｊｅｃｔｉｖｅ　To detect the expression level and secretion change of brain derived neurotrophic factor (BDNF) in rat hippocampus and hippocampal astrocytes induced by propofol and address its mechanism.　Methods　According to the treatment time of propofol, 24 rats were randomly divided into three groups, i.e. 0, 45 min and 90 min groups. Rats were administrated intraperitoneally with propofol(10 g/L, 100 mg/kg body weight). The levels of BDNF mRNA and glial fibrillary acidic protein (GFAP) in rat hippocampus were evaluated by real time polymerase chain reaction (PCR) and immunohistochemistry respectively. Primary cultured hippocampal astrocytes in vitro were divided into 3 groups: Control group (group Ctrl), Propofol group (group Pro), and Inhibitor PD98059+ Propofol group (group Pro-PD). In group Pro-PD, astrocytes were pretreated with 10 μmol/L extracellular signal regulated kinase (ERK) inhibitor PD98059 for 2 h, then incubated with 10 μmol/L propofol for 24 h. Finally cell apoptosis and the expression and secretion levels of BDNF were examined.　Results　Forty-five min and 90 min after injection of propofol, the mRNA levels of BDNF in the hippocampal tissue were 1.20±0.05 fold and 0.86±0.04 of the expression level in group 0 min respectively (P<0.05). Thus, the mRNA level of BDNF were time-dependently altered. At 45 min after injection of propofol, the expression level of BDNF was increased, but at 90 min after injection, the expression level was decreased. The expression level of GFAP in astrocytes detected by immunohistochemistry was also increased after 45 min injection of propofol. In vitro, the survival rates of primary cultured hippocampal astrocytes were (91.2±2.4)% and (90.3±3.0)% in group Ctrl and group Pro respectively. The difference in two groups was not significant(P>0.05). The BDNF mRNA level of hippocampal astrocytes in group Pro was 1.15±0.04 fold of that in group Ctrl (P<0.05). The BDNF mRNA level in group Pro-PD was 0.92±0.05, which was significantly lower than that of group Pro (P<0.05). The BDNF secretion level of hippocampal astrocytes in group Pro was (62±4) ng/L, which was higher than(49±3) ng/L in group Ctrl. The BDNF secretion level in group Pro-PD was (44±3) ng/L, which was significantly lower than that of group Pro(P<0.05).　Conclusions　Propofol activated the rat hippocampal astrocytes and enhanced the expression level and secretion of BDNF in astrocytes through ERK signaling pathway.
Key words: Propofol;　Astrocyte;　Extracellular signal-regulated kinase;　Brain derived neurotrophic factor