【摘要】 目的 探讨原代培养的新生大鼠海马神经元线粒体钙单向转运体(mitochondrial calcium uniporter，MCU)对线粒体通透性转换孔(mitochondrial permeability transition pore,mPTP)是否有调控作用。方法 原代培养的大鼠海马神经元采用随机数字表法分为7组,每组6孔：对照组不预先给药；精胺（Spermine，Sper）组，检测时给予30umol/L的Sper；Ru360组检测前30min给予10umol/L的Ru360；环孢菌素（Ciclosporin A，CsA）组检测前30min给予0.2umol/LCsA；苍术甙（Atractyloside，Atr）组检测时给予400umol/L的Atr；Ru360+Atr组，检测前30min给予10umol/L的Ru360，检测时给予400umol/L的Atr；CsA + Sper组，检测前30min给予0.2umol/L CsA，检测时给予30umol/L的Sper；建立钙超载模型：各个实验组上机检测时都给予200umol/L的CaCl2。在激光共聚焦显微镜下持续监测6min观察各个实验组线粒体calcein AM荧光强度随着时间的变化。结果 对照组线粒体calcein AM荧光强度迅速减低(0.073±0.011)；与对照组比较，Ru360组（0.779±0.015）、CsA组（0.923±0.011）线粒体calcein AM荧光强度减低缓慢（P＜0.05）；与Sper组（0.023±0.006）相比，CsA+Sper组（0.713±0.032）线粒体calcein AM荧光强度减低缓慢（P＜0.05）；Ru360+Atr组（0.643±0.087）与Atr组（0.047±0.015）相比线粒体calcein AM荧光强度减低缓慢（P＜0.05）；与Ru360组（0.779±0.015）相比，Sper组（0.023±0.006）线粒体内Calcein AM的荧光强度的迅速减低（P＜0.05）。 结论 在体外培养的海马神经元水平上MCU对mPTP有调控作用。
【Abstract】 Objective To investigate whether mitochondrial calcium uniporter（MCU） can regulate the mitochondrial permeability transition pore（mPTP） in primary hippocampal neurons. Methods The hippocampal neurons were randomly divided into seven groups according to the Random number table: (1)control group（n=6）: without any prior administration; (2) Spermine group（n=6）: gave 30umol/L Spermine before the test; (3) Ru360 group（n=6）: gave 10umol/L Ru360 30 minutes before the test; (4) Ciclosporin A group（n=6）: gave 0.2umol/L Ciclosporin A 30 minutes before the test; (5) Atractyloside group（n=6）: gave 400umol/L Atractyloside before the test; (6)Ru360+ Atractyloside group（n=6）: gave 10umol/L Ru360 30 minutes before the test and then gave 400umol/L Atractyloside before the test; (7) Ciclosporin A + Spermine group（n=6）: gave 0.2umol/L Ciclosporin A 30 minutes before the test and then gave 30umol/L Spermine before the test. 200umol/L CaCl2 were added to each group at the time of the test began to established the model of calcium overload.Then monitored the mitochondrial calcein AM fluorescence intensity continuously over time for 6 minutes under the laer confocal microscope. Results The mitochondrial calcein AM fluorescence intensity of the control group reduced rapidly(0.073±0.011); compared with the control group, the mitochondrial calcein AM fluorescence intensity of the Ru360 group（0.779±0.015） and Ciclosporin A group（0.923±0.011） reduced slowly(P＜0.05); compared with the Spermine group（0.023±0.006）, the mitochondrial calcein AM fluorescence intensity of the Ciclosporin A + Spermine group （0.713±0.032）reduced slowly (P＜0.05); the mitochondrial calcein AM fluorescence intensity of the Ru360+ Atractyloside group（0.643±0.087） was slower than the Atractyloside group（0.047±0.015）(P＜0.05); the mitochondrial calcein AM fluorescence intensity of the Ru360 group （0.779±0.015）was slower than the Spermine group(P＜0.05). Conclusion Mitochondrial calcium uniporter could regulate the mitochondrial permeability transition pore in hippocampal neurons in vito.
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