CN113624976A - 一种与肝衰竭相关的新型分子诊断标志物组合及其用途 - Google Patents
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Abstract
本发明公开了一种与肝衰竭相关的新型分子诊断标志物组合，该诊断标志物为ATIII和MMR，其诊断效率高，可靠性有保障，能够实现肝衰竭的快速筛查。该诊断标志物可以制备用于诊断肝衰竭或判断肝衰竭预后的试剂或试剂盒，以及可以制备预防或治疗肝衰竭的药物。
Description
技术领域
本发明属于生物技术和肝衰竭生物学技术领域，涉及一种与肝衰竭相关的新型分子诊断标志物组合及其应用。
背景技术
肝衰竭是临床常见的严重肝病症候群，以黄疸、凝血功能障碍、肝性脑病等为主要表现，是临床上病死率极高的一类疾病。肝衰竭可由多种致病因素导致，在我国乙肝病毒感染是肝衰竭主要致病因素，其次是药物和酒精等肝毒性物质。病毒感染导致的肝衰竭具有病情进展迅速、并发症多样、治疗困难、病死率高等特点。早期肝衰竭患者通过去除病因（如服用抗病毒药物或立即停止致病药物摄入）或人工肝支持等综合治疗，可以为肝再生赢得时间，从而使病情得到缓解。中晚期肝衰竭患者唯一有效的治疗手段则是肝移植，然而有限的肝源、昂贵的治疗费用、巨大的手术风险以及术后长期生存率低等因素严重限制了肝移植的广泛应用。因此，如何在疾病早期阻止肝衰竭的发生发展成为迫切需要解决的临床问题。在肝衰竭发生、发展的相关作用机制仍然不完全清楚，但人们对肝病的防治意识逐渐增强，对肝衰竭的早期筛查诊断有了更高的期待。所以一种新型的、特异性强、灵敏度高的能够用于肝衰竭早期筛查，并改善了部分患者预后的肝衰竭分子标记物急需被发现来指导临床的工作。
肝衰竭的典型临床特征之一是凝血因子和抗凝因子的分泌失调，导致凝血功能障碍。抗凝血酶III(ATIII)是一种天然抗凝血剂，仅在肝细胞中合成，能使凝血系统的多种酶失活。据报道，ATIII水平在各种肝脏疾病中降低，如肝硬化、肝炎和妊娠脂肪肝。ATIII活性降低被认为是肝硬化患者肝性脑病的独立预测因子。在原发性肝癌患者中，血清ATIII降低是肝切除术后肝功能障碍的有效预测因子。与这些结果一致，乙肝导致的慢加急性肝衰竭组患者血浆ATIII水平明显降低。
甘露糖受体（MMR）位于巨噬细胞、树突状细胞等多种细胞类型的表面。MMR作为一种模式识别受体，结合并内化各种病原体(如病毒、细菌和寄生虫)的糖蛋白，从而在先天和适应性免疫应答中发挥重要作用。MMR的另一个功能是消除炎症反应过程中释放到循环中的炎症因子。有报道称，伴随慢性丙型肝炎的肝硬化患者血清中可溶性MMR浓度高于轻度肝纤维化患者。同样的，我们的研究表乙肝导致的慢加急性肝衰竭患者血浆MMR明显高于慢性乙型肝炎患者和健康人。由此推测，乙肝导致的慢加急性肝衰竭可显著触发MMR介导的免疫和炎症反应。
研究发现，ATIII和MMR的组合可以有效地区分乙肝导致的慢加急性肝衰竭患者与慢性乙型肝炎患者。ATIII和MMR是新的潜在肝衰竭调控因子，其作为肝衰竭调控因子分别调控炎症凝血功能，有望成为潜在的肝衰竭精准治疗新靶点，具体来讲就是，一种名为ATIII的生物标记物在肝衰竭患者血浆本中低表达，另一种名为MMR的生物标记物在肝衰竭患者血浆本中高表达，提示与肝衰竭患者的预后密切相关，这对于寻找替代性的肝衰竭治疗策略具有十分重要的意义。
发明内容
本发明的第一个目是：针对现有技术之不足，提供一种与肝衰竭相关的新型分子诊断标志物组合，诊断标志物为ATIII和MMR，该诊断标志物组合特异性好，能准确鉴别乙肝导致的慢加急性肝衰患者/慢性乙型肝炎患者；敏感性高，能早期检测出乙肝导致的慢加急性肝衰患者患者且诊断效率高，可靠性有保障，能够实现乙肝导致的慢加急性肝衰患者的快速筛查。
本发明的第二个目的是：提供与肝衰竭相关的诊断标志物的用途。
本发明的第三个目的是：提供诊断肝衰竭、判断肝衰竭预后的试剂或试剂盒。
本发明的第四个目的是：提供预防或治疗肝衰竭的药物。
本发明所采用的技术方案是，一种与肝衰竭相关的新型分子诊断标志物组合，诊断标志物组合为ATIII和MMR。
一种与肝衰竭相关的新型分子诊断标志物组合在制备用于诊断肝衰竭的试剂或试剂盒中的用途。
一种与肝衰竭相关的新型分子诊断标志物组合在制备用于判断肝衰竭预后的试剂或试剂盒中的用途。
进一步地，肝衰竭为乙肝导致的慢加急性肝衰。
一种诊断肝衰竭、判断肝衰竭预后的试剂或试剂盒，试剂或试剂盒中包含ATIII和MMR。
进一步地，肝衰竭为乙肝导致的慢加急性肝衰。
一种与肝衰竭相关的新型分子诊断标志物组合在制备预防或治疗肝衰竭的药物中的用途。
两种预防或治疗肝衰竭的药物，药物中包含ATIII和MMR。
本发明的有益效果是：本发明提供了一种新型的肝衰竭诊断分子标志物组合，其特异性好，能准确鉴别慢加急性肝衰患者/慢性乙型肝炎患者；敏感性高，能早期检测出肝衰竭患者且诊断效率高，可靠性有保障，能够实现肝衰竭的快速筛查。
附图说明
图1是本发明实施例1中乙肝导致的慢加急性肝衰患者（ACLF组）、慢性乙型肝炎患者（CHB组）和健康人（CON组）血浆中ATIII和MMR表达量分析图，其中质谱检测法（MS）检测慢加急性肝衰患者、慢性乙型肝炎患者和健康人各20例，酶联免疫吸附法（ELISA）分别检测慢加急性肝衰患者、慢性乙型肝炎患者和健康人23、45、20例。
图2是本发明实施例2中受试者工作特征曲线（ROC），ROC分析表明，ATIII和MMR的组合预测乙肝导致的慢加急性肝衰具有良好的敏感性和特异性。
具体实施方式
下面结合具体实施例和附图对本发明的技术方案做进一步说明，但应该理解本发明的保护范围并不受具体实施例的限制。
实施例1乙肝导致的慢加急性肝衰患者（ACLF组）、慢性乙型肝炎患者（CHB组）和健康人（CON组）血浆中ATIII和MMR表达量分析
本申请发明人用质谱检测法（MS）检测了慢加急性肝衰患者、慢性乙型肝炎患者和健康人各20例血浆中ATIII和MMR表达量，酶用联免疫吸附法（ELISA）分别检测了慢加急性肝衰患者、慢性乙型肝炎患者和健康人23、45、20例血浆中ATIII和MMR表达量。步骤如下：
1）用抗凝管分别收集慢加急性肝衰患者、慢性乙型肝炎患者和健康人血液,4℃静置4小时，3000 g离心15 min收集血浆，保存于-80度；
2）使用安捷伦多重亲和去除系统(MARS Human-14, Agilent, USA)去除血浆中的14种高丰度蛋白，用胰蛋白酶在37℃消化样品，加入适量的ITRAQ标记后，进行LC-MS/MS检测分析，对获得的质谱数据进行数据导入基于Paragon算法和ProGroup的Protein Pilot（SoftwareVersion 4.5）软件中，进行比对分析。
3）分别使用ATIII ELISA定量试剂盒（R&D systems）检测和MMR ELISA定量试剂盒（RayBiotech）检测血浆中ATIII和MMR表达量。
实验结果如图1所示，ATIII在肝衰竭患者血浆本中低表达，MMR在肝衰竭患者血浆本中高表达。
实施例2ATIII和MMR的组合预测乙肝导致的慢加急性肝衰敏感性和特异性分析
申请发明人根据酶用联免疫吸附法检测测结果，使用SPSS 19.0软件绘制受试者工作特征曲线（ROC），分析ATIII和MMR的组合预测乙肝导致的慢加急性肝衰敏感性和特异性。
实验结果如图2所示，显示ROC分析表明，ATIII和MMR的组合预测乙肝导致的慢加急性肝衰具有良好的敏感性和特异性。
Claims (8)
1.一种与肝衰竭相关的新型分子诊断标志物组合，其特征在于，所述诊断标志物为ATIII和MMR。
2.根据权利要求1所述的与肝衰竭相关的新型分子诊断标志物组合在制备用于诊断肝衰竭的试剂或试剂盒中的用途。
3.根据权利要求1所述的与肝衰竭相关的新型分子诊断标志物组合在制备用于判断肝衰竭预后的试剂或试剂盒中的用途。
4.根据权利要求2或3所述的用途，其特征在于，所述肝衰竭为乙肝导致的慢加急性肝衰竭。
5.一种诊断肝衰竭、判断肝衰竭预后的试剂或试剂盒，其特征在于，所述试剂或试剂盒中包含ATIII和MMR。
6.根据权利要求5所述的诊断肝衰竭、判断肝衰竭预后的试剂或试剂盒，其特征在于，所述肝衰竭为乙肝导致的慢加急性肝衰竭。
7.根据权利要求1所述的与肝衰竭相关的新型分子诊断标志物组合在制备预防或治疗肝衰竭的药物中的用途。
8.一种预防或治疗肝衰竭的药物，其特征在于，所述药物中包含ATIII和MMR。
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| CN116884631B (zh) * | 2023-09-06 | 2023-12-12 | 杭州生奥信息技术有限公司 | 基于ai和相似患者分析的综合肝衰竭预测与治疗参考系统 |
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