【摘要】 目的探讨右美托咪定（DEX）诱导的肌细胞钙浓度双相增加与能量代谢之间的相关性。方法选取出生5天后的雄性Wistar大鼠做为实验中的研究对象,共分为2大组:对照组和右美托咪定组（低剂量0.1μmol/L、高剂量1μmol/L）。利用荧光蛋白钙指示剂测定大鼠肌细胞中钙的变化情况,利用酶标仪测定线粒体琥珀酸脱氢酶反应线粒体的功能,采用1420Victor多标记免疫分析仪测定分析肌细胞中线粒体膜电位的变化,利用高效液相色谱技术测定肌细胞中三磷腺苷、二磷腺苷以及一磷腺苷含量变化,利用流式细胞仪测定肌细胞细胞凋亡情况。采用孔雀绿比色法测定大鼠肌细胞中Ca²⁺-ATPase,Na⁺-K⁺-ATPase酶的活性。利用Western Blot检测大鼠Caspse3表达情况。利用PCR技术检测线粒体周围能量基因表达情况。结果右美托咪定诱导的肌细胞钙浓度双相增加,且右美托咪定诱导浓度越高,钙离子浓度越高,差异有统计学意义（P<0.05）。测定发现右美托咪定作用使得肌细胞线粒体琥珀酸脱氢酶、线粒体膜电位、ATP、ADP、AMP以及Ca²⁺-ATPase,Na⁺-K⁺-ATPase酶的活性均有所增加,且高剂量右美托咪定的作用高于低剂量右美托咪定的作用,而右美托咪定的作用使得肌细胞细胞凋亡率下降,高剂量右美托咪定使得肌细胞细胞凋亡率下降高于低剂量右美托咪定的作用,细胞凋亡中Caspse3基因表达也降低。线粒体周围能量表达相关基因增强,差别有统计学意义（P<0.05）。结论右美托咪定诱导的肌细胞钙浓度双相增加,促使肌细胞能量供应增加,改善肌细胞能量代谢过程。更多还原

【Abstract】 Objective To investigate the relationship between biphasic increase of calcium concentration and energy metabolism induced by dexmedetomidine（DEX）.Methods Five day old male Wistar rats were divided into two groups:control group and dexmedetomidine group（low dose 0.1μmol/L and high dose 1μmol/L）.The changes of calcium in myocytes were measured by fluorescent protein calcium indicator,the function of mitochondrial succinate dehydrogenase reaction mitochondria was measured by enzyme labeling instrument,the changes of mitochondrial membrane potential in myocytes were analyzed by 1420 Victor multi-labeling immunoassay,and the contents of adenosine triphosphate,adenosine diphosphate and adenosine phosphate in myocytes were determined by high performance liquid chromatography.The apoptosis of myocytes was measured by flow cytometry.The activities of Ca²⁺-ATPase,Na⁺-K⁺-ATPase in rat myocytes were determined by malachite green colorimetry.Western Blot was used to detect the expression of Caspse3 in rats.The expression of energy genes around mitochondria was detected by PCR.Results The calcium concentration of muscle cells induced by dexmedetomidine increased,and the higher the induction concentration of dexmedetomidine,the higher the calcium concentration,the difference was statistically significant（P<0.05）.It was found that dexmedetomidine increased the activity of mitochondrial succinate dehydrogenase,mitochondrial membrane potential,ATP,ADP,AMP and Ca²⁺-ATPase,Na⁺-K⁺-ATPase in myocytes,and that the effect of high dose dexmedetomidine was higher than that of low dose dexmedetomidine,while the effect of dexmedetomidine decreased the apoptotic rate of myocytes,and that of high dose dexmedetomidine decreased the apoptotic rate of myocytes.The apoptotic rate of myocytes decreases with dexmedetomidine,which is higher than that of low dose dexmedetomidine.Caspse3 gene expression was also decreased in apoptotic cells.Enhanced expression of energy-related genes around mitochondria.The difference was significant（P<0.05）.Conclusion The biphasic increase of calcium concentration induced by dexmedetomidine can increase the energy supply of myocytes and improve the energy metabolism of myocytes.更多还原