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.     

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.     

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.     

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.     

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.     

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。     

β-Adrenergic receptors and nitric oxide generation in the cardiovascular system

Nitric oxide plays a crucial role in cardiovascular homeostasis, with important vasodilatory, anti-thrombotic and anti-atherogenic properties. β-Adrenergic receptors (βARs), present on a wide variety of cardiovascular cells, including vascular endothelial cells, platelets, cardiac myocytes and leukocytes, have long been established as key players in maintaining cardiovascular homeostatic control. During the last few years a wealth of evidence has emerged which directly links stimulation of these cardiovascular βARs to nitric oxide (NO) generation, suggesting a new and important mechanism of adrenergic control of cardiovascular function. This review explores the cardiovascular cell systems in which this coupling of βARs and NO occurs, the intracellular signalling and regulatory mechanisms involved and the abnormalities in βAR-NO oxide coupling found in cardiovascular disease states. Received 30 September 2005; received after revision 24 November 2005; accepted 24 January 2006     

Analysis of OCT4 expression in an extended panel of human tumor cell lines from multiple entities and in human mesenchymal stem cells

OCT4 is considered a main regulator of embryonic stem cell pluripotency and self renewal capacity. It was shown that relevant OCT4 expression only occurs in cells of embryonic pluripotent nature. However, several recent publications claimed to have demonstrated OCT4 expression in human somatic tumor cells, human adult stem or progenitor cells and differentiated cells.We analysed 42 human tumor cell lines from 13 entities and human bone marrowderived mesenchymal stem cells (MSC). To validate OCT4 expression we used germ cell tumor (GCT) cell lines, derived xenografts and GCT samples. Analysis by RT-PCR, western blotting, immunocytochemistry and immunohistochemistry was performed. With exception of typical embryonal carcinoma cells, we did not observe reliable OCT4 expression in somatic tumor cell lines and MSC. We suggest that a high level of expression of the OCT4 protein together with its nuclear localization still remains a reliable and definitive feature of cells with embryonic pluripotent nature. Received 30 September 2008; received after revision 05 November 2008; accepted 10 November 2008