Gene expression in myocarditis and dilated cardiomyopathy and basic study of gene therapy by plasmid

1)心肌炎和扩张型心肌病的基因表达为研究EAM心肌组织的基因表达，采用实时荧光定量逆转录聚合酶链式反应(RT-PCR)检测免疫后0、14、21和28天EAM大鼠心肌合成的44种基因[钙调节蛋白、收缩蛋白、利钠肽(NPs)、细胞因子、趋化因子、生长因子、肾素-血管紧张素-醛固酮(RAA)系统、内皮素(ETS)和细胞外基质]的绝对拷贝数。在第14天，α-心肌肌球蛋白降低26.3倍，β-心肌肌球蛋白增加3.75倍。心房NP和脑NP在第21天和第14天分别增加47.7倍和6.35倍。血管紧张素III型受体、血管紧张素转换酶和ET1在第21天分别增加22.3倍、6.30倍和16.8倍。在第14天，醛固酮受体减少了2.15倍，但仅在第14天和第21天检测到醛固酮合成酶。白介素2、白介素…More-10、干扰素-γ和单核细胞趋化蛋白-1在第14天分别增加9.08倍、398倍、43.1倍和142倍。21d时，3型胶原、1型胶原和纤维连接蛋白分别增加了34.6倍、1.74倍和44.4倍。有趣的是，骨桥蛋白在第14天时增加了4540倍，是所有基因中最高的。EAM中心肌肌球蛋白和NP的含量发生了显著的变化。RAA系统和ET的表达在EAM的时间进程中会发生不同的变化。细胞因子、趋化因子和细胞外基质大量增加，尤其是早期EAM大量表达骨桥蛋白mRNA。急性炎症愈合后，给予血管紧张素转换酶抑制剂(ACEI)和1型ALL受体阻滞剂治疗。这些药物对血流动力学和心肌收缩性能有良好的影响，可预防纤维化，抑制ANP的表达，逆转心肌肌球蛋白的表型变化。2)裸血浆基因治疗的基础研究大鼠可通过尾静脉快速注射基因进入肝脏，其体积比小鼠大10倍以上，对大鼠的基因功能分析具有重要价值。基于流体力学的pCAGGS-VIL-10基因转移抑制新月体肾炎。PCAGGS-VIL-10大鼠肾小球G6PDH转录本的干扰素-γ、肿瘤坏死因子-α和单核细胞趋化蛋白-1均显著低于pCAGGS大鼠。我们展示了一种实用而简便的方法来检测血液中循环的基因治疗产品，使用胰高血糖素19-29标签载体。较少

1)Gene expression in myocarditis and dilated cardiomyopathyTo investigate gene expression in the myocardium of EAM, absolute copy numbers of 44 mRNA species[calcium-handling proteins, contractile proteins, natriuretic peptides(NPs), cytokines, chemokines, growth factors, renin-angiotensin-aldosterone(RAA) system, endothelins(ETs) and extracellular matrix] in synthesized cDNA from a fixed quantity of total heart RNA were assessed using real-time reverse-transcriptase PCR at days 0,14,21 and 28 after immunization. alpha-Cardiac myosin showed a 26.3-fold decrease and beta-cardiac myosin a 3.75-fold increase at day 14. Atrial NP and brain NP increased 47.7-and 6.35-fold at days 21 and 14 respectively. Angiotensin II type I receptor, angiotensin-converting enzyme and ET1 increased 22.3-fold at day 21,6.30-fold at day 21 and 16,8-fold at day 14 respectively. Aldosterone receptor decreased 2.15-fold at day 14,but aldosterohe synthetase was detected only at days 14 and 21. Interleukin(IL)-2,IL … More -10,interferon-gamma and monocyte chemo-attractant protein-1 increased 9.08-fold at day 14,398-fold at day 21,43.1-fold at day 14 and 142-fold at day 14 respectively. Collagen type 3, collagen type 1 and fibronectin increased 34.6-,1.74-and 44.4-fold respectively at day 21. Interestingly, osteopontin showed a 4540-fold increase and it was the highest mRNA of all at day 14. An isofonn of cardiac myosin and NP are dramatically changed in EAM. RAA system and ET expressions are changed differently during the EAM time course. Cytokine, chemokine and extracellular matrix greatly increase and, in particular, large numbers of osteopontin mRNA are expressed in early EAM. After acute inflammation healed, rats were treated with angiotensin converting enzyme inhibitors(ACEI) and type 1 All receptor blockers. These agents had favorable effects on hemodynamics and myocardial contractility, prevented fibrosis, suppressed the expression of ANP, and reversed phenotypic change of cardiac myosin. All receptor blockers were less effective than ACEI.2)Basic study of gene therapy by naked plasmidGene transfer into the liver via rapid tail vein injection can easily be achieved in the rat, which is more than 10 times larger than the mouse, and has significant value for gene function analysis in rats. pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis. IFN-gamma, TNF-alpha and MCP-1 to the transcripts of G6PDH in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. We demonstrated a useful and convenient method to assay gene therapy products circulating in blood using a glucagon 19-29 tagging vector. Less