Interventional effect of inhibitor of calcium/calmodulin-dependent protein kinaseⅡ on calcium overloading of atrial muscle cells in neonatal rats / 第三军医大学学报

ABSTRACT

Objective To investigate the effect of calcium/calmodulin-dependent protein kinaseⅡ (CaMK[KG-*6]Ⅱ) inhibitor,KN93 on calcium overloading of atrial muscle cells in neonatal rats and detect the expression of CaMK[KG-*6]Ⅱ. Methods The atrial muscle cells from neonatal rats were primarily cultured for 96 h and then divided into 6 group,control,calcium overloading group,KN93 group (0.5 ?mol/L),low-,moderate-and high-dose of KN93+ calcium overloading group. A model of calcium overloading for atrial muscle cells was established by using calcium ionophore (ionomycin,1.0 ?mol/L). For the later 3 groups,KN93 at doses of 0.25,0.5 and 1.0 ?mol/L was added into the culture medium for 30 min followed by 1.0 ?mol/L ionomycin treatment for another 30 min. The identification of ?-actin was performed by immunofluorescence staining. In the present of Fluo-3/AM (an indicator of calcium),intracellular calcium and the expression of CaMK[KG-*6]Ⅱ were detected under the intervention of KN93 with laser cofocal microscopy and Western blotting respectively. Results More than 90% of cultured cells were positive to ?-actin antibody. Compared with the control group,the fluorescence intensity of intracellular Ca2+ was increased significantly by ionomycin (660.16?108.47 vs 376.12?57.57,P