中国医科大学学报2012，Vol.41Issue(12)：1060-1064,5.
部分纯化的牛心细胞质提取物对心肌细胞膜“run-down”L-型钙通道活性的恢复作用
Effects of Partial Purified Cytoplasmic Extract of Bovine Heart to Recover L-type Ca2+ Channel Activity after "Run-down"
摘要
Abstract
Objective To investigate the effects of partial purified cytoplasmic extract and calmodulin-dependent protein kinase Ⅱ (CaMKⅡ ) in the regulation of the activities of L-type Ca2+ channel in myocardial cell membrane of rats. Methods Cytoplasmic extract was pre-pared from bovine heart with routine method, and purified partially wild ion-exchange. Cell attached and inside out mode of patch-clamp technique was employed lo record the single Ca2+ current in the rat ventricular myocytes, and observe the effects of partial purified cytoplasmic extract and CaMK Ⅱ on recovering the activity of L-type Ca2+ channel. Western blot was used to verify the target active protein in the extract. Results Under the inside out mode, the L-type Ca2+ channel showed a "run-down" phenomenon,and its activity decreased to (0.46± 0.15)% recorded under the cell-attached mode. The partial purified cytoplasmic extract recovered L-type Ca2+ channel activity to (44.29± 13.51 )% under the cell-attached mode (P < 0.01 ). After KN-62 (1 μmol/L),a CaMK Ⅱ inhibitor,was added,the L-type Ca2+ channel ac-tivity reduced to (1.30±0.94)%, which was significant different comparing with non-KN-62 adding group( P < 0.05). CaMK Ⅱ was detected in partial purified cytoplasmic extract with western blot method. CaMK Ⅱ prepared with gene-recombination fusion -protein method could also recover "run-down" L-type Ca2+ channel activity to (71.59±14.76)% under the cell-attached mode (P< 0.05). Conclusion The partial purified cytoplasmic extract could recover L-type Ca2+ channel activity after "run-down" whose effect might be related to CaMK Ⅱ activity.关键词
L-型钙通道/钙调蛋白激酶Ⅱ/膜片钳制技术/单通道记录模式Key words
L-type Ca2+ channel/calmodulin-dependent protein kinase Ⅱ /patch-clamp technique/single channel recording分类
医药卫生引用本文复制引用
赵金生,赵妍,赵美眯,刘书源,侯平,郝丽英..部分纯化的牛心细胞质提取物对心肌细胞膜“run-down”L-型钙通道活性的恢复作用[J].中国医科大学学报,2012,41(12):1060-1064,5.基金项目
国家自然科学基金资助项目(30870907,31071004) （30870907,31071004）
