磷酸川芎嗪鼻用pH敏感型原位凝胶对急性脑缺血模型大鼠脑中氨基酸含量的影响

Influence of Nasal Tetramethylpyrazine Phosphate pH-Sensitive in Situ Gel on Amino Acid Content in Rats Brain of Acute Cerebral Ischemia Model

关键词:  高效液相-荧光检测法  磷酸川芎嗪鼻用pH敏感型原位凝胶  氨基酸  脑微透析  急性脑缺血大鼠模型

Key words:  HPLC-FD  tetramethylpyrazine phosphate pH-sensitive in situ gel  amino acid  brain microdialysis  acute cerebral ischemia rat model

摘要:

目的 建立大鼠脑中5种氨基酸浓度的高效液相-荧光检测器测定法(HPLC-FD)，研究磷酸川芎嗪(TMPP)鼻用pH敏感型原位凝胶对急性脑缺血模型大鼠脑部纹状体区氨基酸含量的影响。方法 微透析液用Agilent 1100液相系统荧光检测器检测，ZORBAX SB-Aq C_l8色谱柱，激发波长357 nm，发射波长455 nm；流动相为甲醇和50 mmoL·L^-1乙酸钠缓冲液(pH=6.5)，梯度洗脱，流速为1 mL·min^-1。12只♂大鼠随机分成模型对照组和给药组，每组6只，用微透析采样方法采集大鼠脑部纹状体区微透析液后，HPLC-FD 测定微透析液中5种氨基酸的浓度。结果 鼻腔给予TMPP鼻用pH敏感型原位凝胶(10 mg·kg^-1)后，给药组大鼠体内的Glu和Asp与对照组相比显著下降，而Tau和GABA的量却有所增加。结论 TMPP能明显降低模型大鼠脑内兴奋性氨基酸的含量，增加脑内抑制性氨基酸的含量，对急性脑缺血模型大鼠起到保护作用。

Abstract:

OBJECTIVE To establish a determination method for 5 kinds of amino acid in rats with high performance liquid-fluorescence detector. To study the effects of tetramethylpyrazine phosphate(TMPP) on amino acid content in acute cerebral ischemia rats model. METHODS Microdialysis fluid was determined with Agilent 1100 liquid system fluorescence detector, ZORBAX SB-Aq C_l8 chromatographic column was applied. Excitation wavelength and emission wavelength were set at 357 nm and 455 nm, respectively, mobile phase consisted of methanol and 50 mmol·L^-1 sodium acetate buffer (pH=6.5), with gradient elution, the flow rate was 1 mL·min^-1. Tweleve male rats randomly divided into model control group and treatment group, each group had 6. Microdialysis was used to collect samples in striatum of rats area after dialysate, five kinds of amino acid concentration was determined by HPLC-FD. RESULTS Glu and Asp levels in TMPP group were decreased significantly compare with control group, however, the amount of Tau and GABA increased. CONCLUSION TMPP can obviously reduce excitatory amino acid content and increase brain inhibitory amino acid content in model rats, which has protect effect on acute ceretral ischemia rats.