基于CaN-NFAT3通路探讨充血性心力衰竭常见中医证候本质及潜在标志物

Morbidity of congestive heart failure(CHF) increases by years, and the activation of CaN-NFAT3 pathway is an important pathogenesis of CHF. Study on the essence and markers of common TCM syndromes in CHF is urgent to be broken through. In the preliminary studies, we found that qi-deficiency-blood-stasis syndrome, qi-yin-deficiency syndrome, and yang-deficiency-water-overflowing syndrome are common syndrome patterns of CHF. Moreover, in each type, there is the same differential expression of key proteins in CaN-NFAT3 signal pathway from serum and tongue coating fluid. Based on it, we speculate that the essence and markers of common syndromes in CHF are associated with the key proteins of CaN-NFAT3 pathway. In this study, we will collect chosen patients’ samples from serum, tongue coating fluid, and urine in the 7th, 14th, 21st and 28th days. Then detect the expressions and concentrations of CaN, NFAT3, GATA4 by WB, RIA, and ELISA methods. Then confirm their critical values in diagnosis of different common syndromes by ROC curves. We also will verify the specificity and sensitivity of the values. Furthermore, we will analyze the correlation in the three key proteins and clinical indexes including cardiac function, BNP and LVEF, using rank sum test，Spearman rank correlation，and Delphi methods. In general, we aim to illuminate dynamic relations between this pathway and the essence of CHF common syndromes, to identify and verify the latent markers of CHF common syndromes from multi-sample, and provide theoretical basis for the treatment target of CHF.

充血性心力衰竭（CHF）的发病率逐年增加，CaN-NFAT3通路激活是CHF发生发展的重要机制。CHF常见中医证候本质及标志物的研究亟待突破。我们初步研究发现，气虚血瘀、气阴两虚、阳虚水泛是其常见证型，各证型血清与舌苔液中该通路关键蛋白的差异表达同步，我们推测CHF常见证候的本质及标志物可能与该通路的关键蛋白关联。本项目拟于患者入选第7、14、21、28天收集其血清、舌苔液、尿液标本，应用WB法、RIA法、ELISA法分别检测各标本中该通路关键蛋白CaN、NFAT3、GATA4的表达及其浓度；应用ROC曲线法确定三者用来诊断不同证候的临界值，并验证其特异度和敏感度；应用秩和检验、Spearman等级相关分析结合德尔菲法归纳三者与心功能、BNP、LVEF的临床相关性。阐明该通路与CHF常见证候本质的动态关联，从多标本中明确并印证CHF常见证候的潜在标志物，为本病的中医治疗靶向提供理论依据。

充血性心力衰竭（CHF）的发病率逐年增加，CaN-NFAT3通路激活是CHF发生发展的重要机制。CHF常见中医证候本质及标志物的研究亟待突破。我们前期研究发现，气虚血瘀、气阴两虚、阳虚水泛是其常见证型，各证型血清与舌苔液中该通路关键蛋白的差异表达同步，推测CHF常见证候的本质及标志物可能与该通路的关键蛋白关联。本项目于2018年12月至2020年12月收集CHF气虚血瘀证、气阴两虚证、阳虚水泛证患者（各30例）多时间点血清、尿液、舌苔液样本，应用WB法、RIA法、ELISA法分别检测各标本中该通路关键蛋白CaN、NFAT3、GATA4的表达及其浓度；应用ROC曲线法确定三者用来诊断不同证候的临界值，验证特异度和敏感度；应用秩和检验、Spearman等级相关分析结合德尔菲法归纳三者与心功能、BNP、LVEF的临床相关性。结果发现：①CHF患者血清、尿液、舌苔液中CaN、NFAT3、GATA4表达水平高于健康人群；与阳虚水泛证比较，气虚血瘀证、气阴两虚证中CaN、NFAT3、GATA4蛋白含量降低；CHF常见中医证候在不同时期各蛋白的动态变化趋势一致；同一证候患者血清、舌苔液、尿液中CaN、NFAT3、GATA4蛋白表达呈同步差异变化。②CHF患者血清、尿液、舌苔液中CaN、NFAT3、GATA4蛋白与BNP等临床指标密切相关，其蛋白表达水平及在不同时期含量的变化可直观反映CHF不同中医证候，作为辨别CHF中医证候的内在依据。③多样本CaN-NFAT3通路关键蛋白对CHF常见中医证候的诊断价值均较好。相对于血清、尿液样本中不同蛋白，舌苔液样本CaN对CHF常见中医证候及其不同轻重程度界值的诊断价值较高。通过研究结果，本项目阐明CaN-NFAT3通路与CHF常见证候本质的动态关联，从多标本中明确并印证CHF常见证候的潜在标志物，为本病的中医治疗靶向提供理论依据。