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目的:研究RGK家族中Rem微小蛋白对胚胎小白鼠心室肌细胞Ca~(2+)通道功能的影响。方法:用含有相同浓度正常的Rem(Ad-RemWT)和反转的Rem蛋白(Ad-CTRrem)的两组重组腺病毒(1012/ml)分别感染培养的怀孕18d胚胎小白鼠(ED18)心室肌细胞。12h后,用整个细胞的电压嵌技术来记录ED18心室肌细胞膜L-型Ca~(2+)电流,细胞质Ca~(2+)转运用荧光光度测定法测定。结果:Rem可以很好地在ED18心室肌细胞中表达。在整个细胞的电压嵌技术下,对照组(Ad-CTRrem)能记录到L-型Ca~(2+)电流的细胞为83·3%,实验组(Ad-Rem-WT)为6·6%。Rem有效阻断了ED18心室肌细胞膜Ca~(2+)通道电流。通过荧光光度测定法,测定Rem显著减低了ED18心室肌细胞质Ca~(2+)的转运。结论:外源的RGK家族中Rem蛋白的表达,通过与心室肌细胞Ca~(2+)通道蛋白的β亚单位(Cavβ)相结合改变了Ca~(2+)通道功能,限制了Ca~(2+)的内流。
Objective: To study the effect of Rem protein in the RGK family on Ca 2+ channel function in embryonic mouse ventricular myocytes. Methods: Two groups of recombinant adenovirus (1012 / ml) containing the same concentration of Rem (Ad-RemWT) and inverted Rem protein (Ad-CTRrem) cell. After 12 hours, the L-type Ca 2+ currents of ED18 ventricular myocytes were recorded by whole-cell voltage inlay technique. The cytoplasmic Ca 2+ transport was measured by fluorimetry. Results: Rem was well expressed in ED18 ventricular myocytes. In the whole cell voltage-dependent technique, Ad-CTRrem recorded 83.3% of cells with L-type Ca 2+ currents, while the experimental group (Ad-Rem-WT) %. Rem effectively blocked the ED2 ventricular myocytes Ca2 + channel current. Determination of Rem by fluorescence spectrophotometry significantly reduced Ca 2+ transport in the ventricular myocytes of ED 18 cells. CONCLUSION: The expression of Rem protein in exogenous RGK family changes the function of Ca 2+ channel by binding with the β subunit of Ca 2+ channel protein in ventricular myocytes, (2+) influx.