摘要
目的 :探讨磷酸肌酸 (phosphocreatine ,CP)保护心肌细胞线粒体功能的机制。方法 :采用胶原酶分离成年大鼠心肌细胞 ,0 2mmol/L过氧化氢 (hydrogenperoxide,H2 O2 )刺激细胞 ,于刺激前 10min加入 1mmol/L、10mmol/LCP ,利用荧光探针JC 1结合流式细胞仪检测线粒体膜电位的变化 ;Annexin V/PI及TUNEL染色法鉴定心肌细胞的凋亡。使用 10 μmol/L鱼藤酮及 2 μmol/L碘乙酰胺分别抑制呼吸链及磷酸肌酸激酶 (CK) ,观察其对CP保护作用的影响。结果 :H2 O2 导致线粒体膜电位下降 ,细胞凋亡增加。磷酸肌酸有效地抑制了线粒体膜电位的下降 ,减少了细胞凋亡。鱼藤酮及碘乙酰胺可使其保护作用消失。结论 :CP能够抗心肌细胞氧化损伤 ,其机制可能依赖于减低线粒体膜电位的下降 ,保护线粒体正常功能。
Objective: To observe the protective effect of phosphocreatine on mitochondrial membrane potential in rat cardiomyocytes .Methods: Using H 2 O 2 to cause oxidatie stress, CP was given ten minutes before H 2 O 2 stimulation.JC-1 in combination with flow cytometry was used to measure the changes of mitochondrial membrane potential. Apoptosis was identified by means of double fluorescence staining with Annexin V-FITC / propidim iodide and TUNEL staining. The protective effect of CP was also appraised after rotenone and iodoacetamide (IA) was given to inhibit respiratory chain and CK enzyme respectively. Results: H 2 O 2 caused the decrease of mitochondrial membrane potential and the increase of apoptosis rate. But in the CP treatment group, mitochondrial membrane potential wasn't decreased significantly. This protective effect disappeared after respiratory chain and CK enzyme was inhibited.Conclusion:CP can prevent the cardiomyocyte from oxidative injury. The mechanism may rely on inhibiting the decrease of mitochondrial membrane potential and protecting mitochondrial function.
出处
《军医进修学院学报》
CAS
2004年第1期15-17,共3页
Academic Journal of Pla Postgraduate Medical School
