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了研究EAM心肌中的基因表达，检测了44种mRNA种类[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，使用实时反向荧光定量PCR技术评估了从固定量的心脏总RNA合成的cDNA中的内皮素（ET）和细胞外基质]。免疫后第0、14、21、28天进行PCR检测。在第14天，α-心肌肌球蛋白减少26.3倍，β-心肌肌球蛋白增加3.75倍。在第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 Ⅰ型受体、血管紧张素转换酶和ET 1分别在第21天、第21天和第14天增加22.3倍、6.30倍和16.8倍。醛固酮受体在第14天下降2.15倍，但醛固酮合成酶仅在第14天和第21天检测到。白细胞介素（IL）-2，IL ...更多信息 -10、干扰素-γ和单核细胞趋化蛋白-1在第14天分别增加了9.08倍、21天增加了398倍、43.1倍和142倍。3型胶原、1型胶原和纤维连接蛋白在第21天分别增加了34.6倍、1.74倍和44.4倍。有趣的是，骨桥蛋白显示出4540倍的增加，并且在第14天是所有mRNA中最高的。心肌肌球蛋白和核蛋白的同种型在EAM中发生显著变化。RAA系统和ET表达在EAM时间过程中有不同的变化。细胞因子、趋化因子和细胞外基质大量增加，特别是骨桥蛋白mRNA在早期EAM中大量表达。急性炎症愈合后，给予血管紧张素转换酶抑制剂（ACEI）和1型A11受体阻滞剂治疗。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）裸质粒基因治疗的基础研究通过快速尾静脉注射将基因转入肝脏在大鼠中可以很容易地实现，其体积是小鼠的10倍以上，对大鼠的基因功能分析具有重要价值。基于流体动力学的pCAGGS-vIL-10基因转染抑制新月体肾小球肾炎IFN-γ、TNF-α和MCP-1对肾小球中G6 PDH转录物的作用在pCAGGS-vIL-10大鼠中均显著低于pCAGGS大鼠。我们证明了使用胰高血糖素19-29标记载体来测定血液中循环的基因治疗产物的有用且方便的方法。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