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研究EAM心肌中44种mRNA的绝对拷贝数[钙处理蛋白、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，在免疫后第0、14、21和28天，采用实时逆转录酶PCR技术检测从固定数量的心脏总RNA合成的cDNA中的内皮素（ETs）和细胞外基质。第14天α -心肌肌球蛋白降低26.3倍，β -心肌肌球蛋白升高3.75倍。第21天和第14天，心房NP和脑NP分别增加47.7倍和6.35倍。血管紧张素II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。醛固酮受体在第14天下降2.15倍，醛固酮合成酶仅在第14和21天检测到。白细胞介素(IL)-2、IL…More -10、干扰素- γ和单核细胞化学引诱蛋白1在第21天分别增加了9.08倍、14398倍、43.1倍和142倍。第21天，3型胶原蛋白、1型胶原蛋白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天增加了4540倍，是所有mRNA中最高的。心肌肌球蛋白和NP的异构体在EAM中发生了显著变化。RAA系统和ET的表达在EAM时间过程中发生了不同的变化。细胞因子、趋化因子和细胞外基质显著增加，尤其是骨桥蛋白mRNA在EAM早期大量表达。急性炎症愈合后，给予大鼠血管紧张素转换酶抑制剂（ACEI）和1型All受体阻滞剂。这些药物对血流动力学和心肌收缩力有良好的影响，防止纤维化，抑制ANP的表达，逆转心肌肌球蛋白的表型改变。2)裸质粒经尾静脉快速注入肝内基因治疗的基础研究在大鼠身上很容易实现，其体积是小鼠的10倍以上，对大鼠基因功能分析具有重要价值。流体动力学转染pCAGGS-vIL-10基因转染抑制月牙体肾小球肾炎。pCAGGS- vil -10大鼠肾小球中ifn - γ、tnf - α和MCP-1对G6PDH转录本的表达均显著低于pCAGGS大鼠。利用胰高血糖素19-29标记载体，对血液中循环的基因治疗产物进行检测。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