咪达唑仑对N-甲基-D-天冬氨酸致伤PC12细胞氨基酸水平的影响  

Effect of midazolam on amino acid levels in cultured PC12 cells challenged by N-methyl-D-aspartate

作　　者：王恒林[1] 王卓强[1] 刘秀珍[1] 曹江北[2] 王永安[3] 李云峰[3] 米卫东[2] 张宏[2] 

机构地区：[1]解放军总医院第二附属医院麻醉科,北京100091 [2]解放军总医院麻醉科 [3]军事医学科学院毒物药物研究所

出　　处：《解放军医学杂志》2008年第1期75-76,共2页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金面上项目(30400551、30500482)

摘　　要：目的观察静脉麻醉药咪达唑仑(MID)对N-甲基-D-天冬氨酸(NMDA)致PC12细胞损伤时细胞氨基酸水平的影响,以探讨MID发挥其细胞保护作用的可能机制。方法建立NMDA(300μmol/L)诱导的PC12细胞损伤模型。将培养好的PC12细胞随机分为对照组、NMDA(300μmol/L)组、MID组(其中又分为3μmol/L和30μmol/L亚组),处理4h后收集细胞、漂洗、超声匀浆,4℃离心(12000r/min×20min),取上清用高效液相色谱分析法测定PC12细胞内氨基酸含量。结果NMDA 300μmol/L处理4h可使PC12细胞谷氨酸含量显著增加,但天冬氨酸、谷氨酰胺、甘氨酸含量无明显改变。MID3μmol/L、30μmol/L分别与NMDA 300μmol/L同时处理PC12细胞4h后,谷氨酸含量较NMDA(300μmol/L)组明显降低(P<0.05),而对天冬氨酸、谷氨酰胺、甘氨酸则无明显影响。结论NMDA 300μmol/L可显著增加PC12细胞的谷氨酸水平而导致细胞损伤,而MID可抑制NMDA诱导PC12细胞损伤所致的谷氨酸释放,提示MID可能是通过抑制谷酸的释放而发挥其保护细胞的作用。Objective To observe the effect of rnidazolam （MID） on amino acid levels in cultured PC12 cells challenged by N-methy-D-aspartate （NMDA）, and to explore the possible protective action of midazolarn which is an anesthetic, Methods Cultured PC12 cells were divided into control group, NMDA group, and MID group. In NMDA group, PC12 cells were challenged by 300μmol/L NMDA in vitra In MID group, 3μmol/L or 30μmol/L of MID was added to the challenged PC12 cell culture, thus forming two subgroups. After being treated with NMDA 300μmol/L for 4 hours, the PC12 ceils were collected, rinsed, levigated and centrifuged （12 000r/rnin for 20min, at 4℃ ）, then the supematant liquid was collected. The levels of amino acids were determined with high performance liquid chroma tograpy （HPLC）. Results After exposing to NMDA 300/μmol/L for 4 hours, the level of glutamate released from PC12 cells rose significantly, whereas the level of glutarnine, aparatate and glycine remained unchanged. In the presence of MID 3μmol/L and 30μmol/L for 4 hours, the level of glutamate was lowered significantly （P〈0. 05, compared with NMDA-treated group）, whereas the levels of glutamine, aparatate and glysine were not significantly changed. Condusions These results suggest that NMDA 300μmol/L can inflict significant impairment on PC12 cells, and this activity may be related to the increase of glutamate. MID in a concentration of 3μmol/L or 30μmol/L can significantly inhibit the NMDA-induced release of glutamate from PC12 cells, suggesting that MID can antagonize the activity of NMDA on PC12 cells, and this protective activity of MID may be related to its inhibitory effect on the release of glutamate.

关 键 词：咪达唑仑 兴奋性氨基酸类 N甲基天冬氨酸 PC12细胞 

分 类 号：R971.2[医药卫生—药品]