声动力治疗中钙超载机制的研究

Mechanisms of the Occurrence of Calcium Overload in Sonodynamic Treatment

目的探讨声动力诱导C6胶质瘤细胞凋亡中钙超载现象及部分机制。方法将对数期生长的C6胶质瘤细胞(4×104个/ml)放入试管中,每管5 ml。实验分对照组、HMME、超声组和声动力治疗组。采用流式细胞仪检测细胞凋亡,荧光分析仪检测细胞内钙离子水平,加入钙离子拮抗剂观察凋亡率的变化;采用免疫印迹的方法观察各组中钙离子ATP酶(SERCA)的表达水平,并分析钙离子的来源。结果声动力治疗组细胞内钙离子浓度为(224.26±10.24)nm,较其他组明显升高,加入钙离子拮抗剂后其细胞凋亡率明显降低,声动力组SERCA2表达明显降低。结论声动力诱导部分作用机制是细胞内出现钙超载现象,其原因可能是声动力对内质网SERCA2的破坏,进一步导致钙离子水平升高,引起凋亡反应所致。

Objective To study the phenomenon and mechanisms of calcium overload occurred in connection with the C6 cell apoptosis induced in SDT. Methods A C6 cell line was maintained in RPMI-1640 medium.The apoptosis rate（23.56±2.78）% was measured with a flow cytometry and the concentration of intracellular Ca2＋ with a fluorometer.The change to the apoptosis rate was observed after calcium antagonist being applied in the SDT.Moreover,SERCA2s in different groups were observed with immunoblotting,and the source of calcium was identified. Results The study indicated that the rise of i in the SDT group is much more dramatic than in other groups,and the apoptosis rate dropped more remarkably in the SDT group upon the application of BAPTA-AM. Conclusions As to C6 cells,calcium overload plays certain roles in SDT-induced cell apoptosis.The mechanisms is that Endoplasmic Reticulum（ER） may be a major target of SDT,which induces SERCA2 degradation and causes further i elevation.