摘要
目的:研究川芎嗪(TMP)对海马神经元细胞膜L型钙通道电流(Ica.L)作用和对神经细胞内钙([Ca2+]i)的影响。方法:取新生24h内大鼠进行原代海马神经元培养,使用全细胞膜片钳技术联合激光扫描共聚焦显微镜观察TMP对神经元Ica.L和[Ca2+]i的影响。结果:①膜片钳证实,10、30、100μmol/L的TMP组能显著抑制Ica.L峰值,分别与对照组的(454.2±31.4)pA比较降至(276.4±17.1、209.3±14.2和(135.8±16.0 pA,P<0.05),使I-V曲线上移,且具有浓度依赖性,但对最大激活电位无明显影响。②激光扫描共聚焦显微镜测量发现,细胞外液有钙液时,10、30和100μmol/L TMP组能显著抑制60 mmol/L的KCL引起的细胞内钙荧光峰值(Fi)增加,与对照组的(1749.4±61.0)比较降至(1227.1±36.2、998.2±23.9和745.9±20.6,P<0.05);在细胞外液无钙时,30μmol/L TMP组也能显著抑制60 mmol/L KCL引起的[Ca2+]i)库释放,与对照组(965.3±82.5)比较降至(789.6±75.6,P<0.05)。结论:TMP具有对海马神经元钙通道Ica.L和[Ca2+]i库释放的双重抑制作用,使[Ca2+]i水平降低,这可能是其神经保护机制原因之一。
Objective：To study the effect of tetramethylpyrazine （TMP） on L-type ＂Calcium-channel current＂ and the intracellular calcium concentration （[Ca^2＋]i） of hippocampal neuronal cells. Methods：Primary culture of hippocampal neuronal cells was performed. Cell patch clamp technique was used to record the current of L-Type calcium channels in hippocampal neuronal cells, and [Ca^2＋ ]i was measured by laser scanning confocal microscope. Results： （1） It was conformed by patch clamp technique that TMP with concentrations of 10, 30 and 100 μmol/L could inhibit Ica. L concentration-dependently, the peak values of Ica. L were reduced to （276.4±17.1）, （209.3±14.2） and （135.8±16.0） pA respectively as compared with control group （454.2±31.4 pA, P〈0.05）, and the I-V curves of Ica. L shifted upwardly. （2） Three different concentrations of TMP could reduce the elevation of [Ca^2＋ ]i induced by 60 mmol/L KCL solution, and could inhibit the release of calcium ion from intracellular calcium store also. Conclusion： Both Ica. L channel and the release of intracellular calcium store can be inhibited by TMP, which might be one of its nerve protective mechanisms.
出处
《中国康复》
2008年第1期17-19,共3页
Chinese Journal of Rehabilitation
关键词
川芎嗪
膜片钳
激光扫描共聚焦显微镜
钙通道
海马神经元
tetramethylpyrazine
patchclamp technique
laser scanning confocal microscope
L-type calcium-channel current
hippocampal neuronal cells
