重组GATA4腺病毒的构建及心肌细胞感染

利用AdEasy腺病毒表达系统构建了GATA4基因过表达腺病毒.编码大鼠GATA4基因的目的片段克隆入pAdTrackcmv质粒,pAdTtrackcmv-GATA4穿梭质粒经PmeⅠ线性化后转入含pAdEasy-1的BJ5183菌进行同源重组.pAdEsay-GATA4重组质粒经卡那霉素抗性筛选及PacⅠ酶切鉴定.pAdEsay-GATA4转入293A细胞进行包装,产生具有感染性的重组病毒.Ad-GATA4重组病毒感染HeLa及乳鼠心肌细胞,通过免疫印迹及实时定量聚合酶链反应(Real-time PCR)检测GATA4表达及促心肌肥大效应.Ad-GATA4重组病毒感染乳鼠心肌细胞后,诱导心肌细胞表面积明显增加,ANF表达明显增强.结果表明,Ad-GATA4腺病毒成功感染心肌细胞并诱导了大鼠心肌肥大表型的出现.     

Gene product expression of cyclin D<Subscript>2</Subscript> and p16 during the transition from cardiac myocyte hyperplasia to hypertrophy

The current study was to investigate mRNA expression of cyclin D₂ and p16 during the transition from cardiac myocyte hyperplasia to hypertrophy. Cultured cardiac myocytes (CM) and fibroblasts (FC) obtained from 1-day-old Sparague-Dawley rats were used in this study. We have determined (1) hyperplasia by cell growth curve and fluorescence activated cell sorting (FACS); and (2) ultrastructure by electron microscope observation; and (3) expresions of cyclin D₂ mRNA and p16 mRNA by using in situ hybridization and image analysis. The results were shown (1) Results of cell growth curve and FACS analysis showed CM could proliferate in the first 3 cultured days (4 days in postnatal development). But the ability decreased quickly, concomitant with the differentiation. (2) The ultrastructure of CM showed the large amount of myofilaments and mitochondrion and FC showed moderate amount of rough endoplasmic reticulum. (3) The expression of cyclin D₂ mRNA in 3−, 4−, 5−day CM group was 0.89 times (p<0.05), 0.80 times (p<0.05) and 0.56 times (p<0.01) of that in 1-day group respectively. P16 mRNA in 2−, 3−, 4−, 5−day CM group were 1.63 times (p<0.01), 1.72 times (p<0.01), 1.99 times (p<0.01) and 2.84 times (p<0.01) of that in 1−day group respectively. It can be concluded that cultured neonatal rat cardiac myocytes could proliferate during the first 3 cultured days, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 have the key roles during the transition from myocyte hyperplasia to hypertrophy. Biography: Zhang Yu-xia (1974-), female, Master, research direction: cardiovascular pathology.     

Cell-to-cell coupling studied by diffusional methods in myocardial cells

Summary The diffusion of large molecular substances from cell to cell in multicellular and enzymatically isolated cell pairs is described. Permeability of the gap junctional membrane to these molecules and the critical diffusing diameter of the myocardial gap junctional channel are discussed.     

Se-scFv-B3对过氧化氢诱导的大鼠乳鼠心肌细胞损伤的保护作用

用H₂O₂损伤大鼠乳鼠的心肌细胞建立氧化应激损伤模型, 考察谷胱甘肽过氧化物酶模拟物Se-scFv-B3对H₂O₂诱导的大鼠乳鼠心肌细胞氧化损伤的影响. 结果表明, Se-scFv-B3能部分增加心肌细胞存活率, 减少细胞凋亡, 恢复线粒体膜电位, 下调Caspase-3活力并降低细胞内活性氧的含量. 表明Se-scFv-B3可以保护心肌细胞抵制H₂O₂诱导的氧化应激损伤.     

Effect of Calcium on Spontaneous Quantal Transmitter Secretion from ACh-Loaded Myocytes

Introduction Transmitter secretion requires specialized secretory or- ganelles, the synaptic vesicles, for the packaging, stor- age, and exocytotic release of the transmitters[1,2]. The neurotransmitter acetylcholine (ACh) is released at the neuromuscular…     

New dynamic model for non-Fickian diffusion of calcium spark in cardiac myocytes

A new dynamic model for non-Fickian diffusion of calcium spark in cardiac myocytes was developed by introducing time lags on the basis of the microscale mass transport theory. Numerical simulation showed that the size of the calcium spark produced by the new dynamic model was larger than that of Fick diffusion and was in more agreement with experimental results. In addition, the time lags of the calcium spark in cardiac myocytes were about 0.1--0.8 ms. These results can be used to understand the mechanism of calcium spark diffusion in cardiac myocytes.     

心室肌细胞持续性钠电流的研究现状

心室肌细胞存在持续性钠电流的特征与心脏其他类型的钠通道电流不同。某些药物,如利多卡因、奎尼丁、藜芦宁、R56865,Anthopleurin,TTX等）及病理、生理状态可影响此电流。此外,该电流可影响动作电位形态,加强心室肌细胞的去极化,并与早期后除极相关,参与心律失常的形成。对INa.p所引起的不同反应及其发生机制的研究,有助于了解机体内外环境紊乱时心律失常的发生机制。     

Gene Product Expression of Cyclin D2 and p16 During the Transition from Cardiac Myocyte Hyperplasia to Hypertrophy

The current study was to investigate mRNA expression of cyclin D2 and p16 during the transition from cardiac myocyte hyperplasia to hypertrophy. Cultured cardiac myocytes (CM) and fibroblasts (FC) obtained from 1-day-old Sparague-Dawley rats were used in this study. We have determined (1) hyperplasia by cell growth curve and fluorescence activated cell sorting (FACS); and (2) ultrastructure by electron microscope observation; and (3) expressions of cyclin D2 mRNA and p16 mRNA by using in situ hybridization and image analysis. The results were shown (1) Results of cell growth curve and FACS analysis showed CM could proliferate in the first 3 cultured days (4 days in postnatal development). But the ability decreased quickly, concomitant with the differentiation. (2) The ultrastructure of CM showed the large amount of myofilaments and mitochondrion and FC showed moderate amount of rough endoplasmic reticulum. (3) The expression of cyclin D2 mRNA in 3-, 4-, 5-day CM group was 0.89 times(p<0.05), 0.80 times (p<0.05)and 0.56 times (p<0.01)of that in 1-day group respectively. P16 mRNA in 2-, 3-, 4-, 5-day CM group were 1.63 times(p<0.01),1.72 times(p<0.01),1.99 times (p<0.01)and 2.84 times (p<0.01) of that in 1-day group respectively. It can be concluded that cultured neonatal rat cardiac myocytes could proliferate during the first 3 cultured days, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D2 and p16 have the key roles during the transition from myocyte hyperplasia to hypertrophy.     

异鼠李素对心肌细胞缺血再灌注损伤的保护作用

本研究在体外建立心肌细胞缺血再灌注模型.利用透射电镜、MTT,流式细胞技术,及乳酸脱氢酶活性检测细胞活性和Western Blot分析等方法探讨异鼠李素对心室肌细胞缺血再灌注损伤的作用.发现缺血再灌注心室肌细胞出现明显的细胞凋亡现象,电镜观察细胞超微结构发生明显改变,MTT,流式细胞技术,及乳酸脱氢酶活性结果显示I/R+/isor+组心肌细胞活性与I/R+/isor相比明显增强,细胞凋亡程度减弱;Western Blot也表明与I/R+/isor组相比,I/R+/isor+组心室肌细胞中Bcl-2蛋白表达明显上调,Bax及p53蛋白表达明显下降, Bcl-2/Bax比率显著减小.从实验结果显示异鼠李素对心肌缺血再灌注损伤具有一定的保护作用,其作用与药物浓度有关,低浓度的异鼠李素对心肌具有保护作用,且药物浓度越大,保护作用越强;而高浓度的异鼠李素则随药物浓度的增加对心肌的保护作用逐渐减弱,甚至出现一定的细胞毒性.     

SD乳鼠心肌细胞的培养及其低温下钙波-搏动耦联的初步研究

分离培养3 d以内SD乳鼠心肌细胞,Fluo-4负载后分别于37 ℃,24 ℃环境中在激光扫描共焦显微镜 (Laser Scanning Confocal Microscopy, LSCM)下进行细胞搏动及胞内Ca2+运动的观测.结果显示,心肌细胞在体外呈集落生长.当37 ℃时,集落中无钙波产生,但集落细胞同步搏动;当24 ℃时,集落中个别细胞先发生钙波,随后集落细胞才同步搏动.证明心肌细胞搏动与胞内Ca2+浓度变化相关,在24 ℃低温下个别细胞产生的钙波可能对其所在集落细胞的同步搏动有诱导并维持其搏