CN113215249B - Il-26用于焦虑抑郁症的鉴别与诊断 - Google Patents
Il-26用于焦虑抑郁症的鉴别与诊断 Download PDFInfo
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Abstract
本发明涉及生物医药领域,具体涉及IL‑26用于焦虑抑郁症的鉴别与诊断。本发明提供一种焦虑抑郁症诊断或鉴别诊断试剂,所述试剂包括检测IL‑26表达水平的物质。本发明通过大量临床样本的证明了IL‑26在焦虑抑郁症患者低表达,作为焦虑抑郁症诊断标记物,具有较高的诊断价值,也可作为区分焦虑抑郁症、精神分裂症和痴呆中的两两疾病间的辅助指标,为基因治疗、药物治疗等临床应用提供了治疗靶点和重要依据。
Description
技术领域
本发明涉及生物医药领域,具体涉及IL-26用于焦虑抑郁症的鉴别与诊断。
背景技术
神经症又可以称之为神经官能症或精神神经症,是对一组精神障碍疾病的总称,如神经衰弱、强迫症、焦虑症、恐怖症、躯体形式障碍等。神经症临床表现的主要特点为患者常深感痛苦,影响心理功能或社会功能,无任何器质性病理基础,大多会出现精神和心理异常,有时还可能会导致心脏、胃肠或生殖系统发生病变,会持续迁延发作。
临床上经常用一个名词叫神经症,非常广泛,也叫神经官能症,或精神神经症,不是一个单独疾病,是一组疾病的总称。包括神经衰弱、焦虑状态、抑郁状态、躯体障碍,是多种疾病综合的总称。临床表现非常复杂,病人非常痛苦,有各种各样的临床表现,但做客观检查时,又检查不出相应器质性疾病,或者检查器质性疾病,不能解释病人目前表现。
焦虑抑郁症(焦虑抑郁障碍)又称焦虑性神经症,是以广泛性焦虑症(慢性焦虑症)和发作性惊恐状态(急性焦虑症)为主要临床表现,常伴有头晕、胸闷、心悸、呼吸困难、口干、尿频、尿急、出汗、震颤和运动性不安等症,其焦虑并非由实际威胁所引起,或其紧张惊恐程度与现实情况很不相称。
据流行病学调查和医疗机构调查报道,近40%~70%的抑郁障碍现患社区人群或机构就诊群体共病至少一种焦虑障碍。国内研究报道,抑郁障碍就诊患者共病焦虑障碍的比例高达60.2%~68.9%,社区人群中心境障碍和焦虑障碍的共患率高达63%。相比于无焦虑障碍共病的抑郁障碍病患,共病群体具有总体疾病特征更复杂、病情更严重、社会经济状况较低下、功能损害更严重、残疾程度更高、生活质量较差等特征。更为重要的是,大量研究结果显示该群体具有更显著的预后不良倾向。
在中国5个城市15家三级甲等综合医院的心血管科、消化科、神经科和妇科门诊,对8487例患者开展诊断性调查显示,焦虑障碍的患病率8.6%,焦虑和抑郁同时存在的患病率是4.1%;445例符合焦虑障碍诊断标准的患者中,接近半数与抑郁障碍共病。针对美国普通人群的共病率研究中,12个月内患有抑郁症且同时诊断为焦虑的比例高达51.2%-57.5%。
一项2009年发布的中国前瞻性、多中心、队列研究共纳入508例符合美国DSM-IV抑郁症诊断标准的患者,采用17项汉密尔顿抑郁量表和焦虑量表、社会功能缺陷筛选量表和临床总体印象量表(CGI)对其进行评估,观察流行病学资料、焦虑抑郁症状群及自杀状况。结果显示,抑郁症患者焦虑障碍的共病率为68.9%,16.7%共病多种焦虑障碍。焦虑障碍种类分布以广泛性焦虑障碍为主,为56.1%。焦虑对抑郁症的临床严重程度有显著统计学意义。36%患者同时伴有躯体疾病。
临床实践中,针对抑郁障碍伴焦虑患者,可选用同时拥有抑郁和焦虑障碍适应证的抗抑郁药物。另外,从抗抑郁药起效机制出发,也可考虑采用5-HT1A受体部分激动剂进行增效治疗,针对焦虑性抑郁障碍患者的疗效似乎应该更为理想,加之长期安全性相对较好,或可作为较优的治疗选择。
但是,焦虑抑郁症发病机制复杂,涉及多系统、多环节的功能失调,可表现为多种生化物质的动态平衡异常。但令人遗憾的是,焦虑抑郁症的病因至今尚不完全清楚,并且对于焦虑抑郁症的诊断还是以临床表现及量表作为主要的诊断依据,目前,尚缺乏焦虑抑郁症诊断及与其它精神神经疾病鉴别诊断相关的生物标志物等客观指标。
发明内容
为了解决上述技术问题,本发明目的在于提供了IL-26蛋白作为焦虑抑郁症、精神分裂症和痴呆中至少一种疾病的诊断标志物及其在产品中的应用。经研究发现,IL-26在焦虑抑郁症患者的血清中显著降低,其作为焦虑抑郁症诊断生物标记物具有较高的灵敏度、特异性和准确度,且能有效区分区分焦虑抑郁症与其它精神神经疾病,其它精神神经疾病包括精神分裂症和痴呆;也能有效区分精神分裂症和痴呆。
为了实现上述目的,本发明采用如下技术方案:
本发明的第一方面提供了一种焦虑抑郁症或精神分裂症诊断试剂,所述试剂包括检测IL-26表达水平的物质。
进一步,与正常组相比,焦虑抑郁患者血清中IL-26显著降低。
进一步,所述试剂选自:识别IL-26的探针;或扩增IL-26引物;或结合IL-26蛋白的抗体。
本发明的第二方面提供了以上所述的试剂在制备诊断焦虑抑郁症产品中的应用。
本发明的第三方面提供了一种区分至少两种精神神经疾病的诊断试剂,所述试剂包括检测IL-26表达水平的物质。
优选地,所述精神神经疾病包括焦虑抑郁症、精神分裂症和痴呆。
优选的,所述检测包括基因水平的检测和蛋白水平的检测。如,所述基因水平的检测包括real-time PCR法、Northern blot、Southern blot、基因芯片、原位杂交或RNA酶保护实验等。所述蛋白水平的检测包括免疫检测、Western blot或蛋白芯片等。
进一步,与正常组相比,焦虑抑郁患者血清IL-26显著降低;精神分裂患者较正常组显著降低,但明显低于焦虑抑郁患者;痴呆患者与正常组无显著差异;焦虑抑郁患者与痴呆患者相比,显著降低。
进一步,所述试剂选自:识别IL-26的探针;或扩增IL-26引物;或结合IL-26蛋白的抗体。
本发明的第四方面提供了所述的试剂在制备区分焦虑抑郁症、精神分裂症和/或痴呆的诊断产品中的应用。优选地,用于包括区分焦虑抑郁症、精神分裂症和/或痴呆的两种或三种疾病。
进一步,所述产品包括制剂、试纸、芯片或试剂盒。
所述试剂盒包括蛋白检测试剂盒如ELISA检测试剂盒、胶体金检测试剂盒、免疫共沉淀试剂盒、化学发光试剂盒、免疫荧光试剂盒等。所述试剂盒中一般配备有相应的使用说明书,说明书一般包括公司标志及名称、试剂盒名称、试剂盒组成、保质期、使用领域、使用方法等项目,用户按照说明书不需或只需少量的优化即可得到满意的结果。
所述ELISA检测试剂盒,所述的试剂盒包括:包被IL-26单克隆抗体的固相载体,酶标抗体,酶的底物,蛋白标准品,阴性对照品,稀释液,洗涤液,酶反应终止液等。
本发明的第五方面提供了能够降低血液中IL-26含量的物质用于制备预防或治疗焦虑抑郁症药物中的应用。
进一步,所述药物包括特异性IL-26基因敲低系统和/或IL-26蛋白抑制系统。
在一些实施方式中,所述基因敲低系统包括shRNA,siRNA和CRISPR/Cas系统;所述shRNA和siRNA基因敲低系统是在RNA水平上降低蛋白的表达,CRISPR/Cas系统是在DNA水平上降低蛋白的表达。
在一些实施方式中,所述蛋白抑制系统包括特异性抗体阻断系统和小分子多肽阻断系统,是在蛋白水平抑制产物的效应。
有益效果
本发明通过大量临床样本的证明了IL-26在焦虑抑郁症患者低表达,作为焦虑抑郁症诊断标记物,具有较高的诊断价值,也可作为区分焦虑抑郁症、精神分裂症和痴呆两两疾病间的辅助指标,为基因治疗、药物治疗等临床应用提供了治疗靶点和重要依据。
附图说明
图1IL-26在任务前、后人员血清中的IL-26荧光强度。
图2IL-26在正常人、焦虑抑郁、精神分裂和痴呆患者血清中的IL-26荧光强度。
图3IL-26诊断焦虑抑郁vs正常人;精神分裂vs正常人;痴呆vs正常人的ROC曲线组合图。
图4IL-26诊断焦虑抑郁vs精神分裂症ROC曲线。
图5IL-26诊断焦虑抑郁vs痴呆的ROC曲线。
图6IL-26诊断精神分裂症vs痴呆的ROC曲线。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。若未特别指明,实施例中所用的技术手段为本领域技术人员所熟知的常规手段,所用的试剂可以商业购买得到。
实施例中未注明具体条件的实验方法,通常为本领域常规方法。
实施例1细胞因子芯片筛选差异蛋白
对执行某长期(7个月)任务中面对较大精神心里压力的青年男性,分别于执行任务前、后,2012年4月至11月期间解放军总医院获取所有对象的肘正中静脉血5ml,室温静置30min,1000×g离心15min,收集血清(其中执行任务前对照样品8例,编号首字母为B;任务后样品12例,编号首字母为X)。利用Raybiotech公司AAH-BLG-1细胞因子芯片,从507个因子中筛选目标蛋白。读出结果为荧光强度,经过标准化后,数值越高代表蛋白浓度越高。芯片结果显示,与对照组相比,在执行任务后的血清中IL-26显著降低(t检验p=0.0138)(图1),拟把IL-26作为焦虑抑郁指征的检测指标。具体数据如表1和表2所示。
表1执行任务前对照组IL-26荧光强度
样品名称 | B54 | B48 | B59 | B58 |
IL-26 | 14263 | 12974.52 | 16138.43 | 15476.55 |
样品名称 | B38 | B30 | B37 | B36 |
IL-26 | 11001.75 | 11208.15 | 11597.58 | 12251.52 |
表2执行任务后实验组IL-26荧光强度
样品名称 | X85 | X193 | X182 | X183 | X110 | X120 |
IL-26 | 7507.623 | 9989.454 | 8005.582 | 8989.697 | 13732.54 | 10349.67 |
样品名称 | X123 | X127 | X173 | X174 | X175 | X176 |
IL-26 | 11364.83 | 17087.611 | 7581.111 | 7426.61 | 1250.599 | 1311.19 |
实施例2临床验证
取2019年6月至2019年8月解放军总医院第一医学中心就诊的抑郁焦虑症候群患者20例,2020年10月成都市第四人民医院就诊的抑郁焦虑症候群患者19例,共39例;同时收集2020年10月成都市第四人民医院就诊的精神分裂症患者20例、痴呆患者14例作对照,健康人对照来源于2019年6月至2019年8月解放军总医院第一医学中心体检但各项指标均正常的志愿者,共20例。获取所有对象的肘正中静脉血5ml,室温静置30min,1000×g离心15min,收集血清做细胞因子芯片。
结果如图2所示,与正常组相比,焦虑抑郁患者血清IL-26显著降低(p=1.13784E-05);精神分裂患者较正常组显著降低(p=1.08049E-06),但明显低于焦虑抑郁患者(p=0.0002);痴呆患者与正常组无显著差异(p=0.524793);焦虑抑郁患者与痴呆患者相比,p<0.0001。各组IL-26浓度结果如表3所示。
表3各组IL-26荧光强度
上述实验说明,IL-26可以作为焦虑抑郁、精神分裂症的诊断标志物,也可作为区分焦虑抑郁症、精神分裂症和痴呆两两疾病间的辅助指标。
使用Graphpad Prism 8软件制受试者工作曲线(ROC),分析AUC值、敏感性和特异性,判断IL-26的焦虑抑郁诊断效能,和判断IL-26在焦虑抑郁、精神分裂症和痴呆两两疾病之间的鉴别诊断效能。其中,AUC越接近于1,则认为诊断价值越好,一般AUC>0.5则认为其具有诊断价值。使用IL-26荧光强度进行分析,选择最大的Youden指数=(敏感度+特异性-1)对应的点水平作为其最佳cutoff值,以此确定特异性和敏感度。
结果显示,IL-26诊断焦虑抑郁的AUC为0.81,p=0.0001313,特异性0.7,灵敏度0.92;IL-26诊断精神分裂症的AUC为0.93,p<0.0001,特异性0.9,灵敏度0.95(图3);IL-26鉴别诊断焦虑抑郁与精神分裂症的AUC为0.84,p<0.0001,特异性0.8,灵敏度0.82(图4);IL-26鉴别诊断焦虑抑郁与痴呆的AUC为0.75,p=0.006888,特异性0.57,灵敏度1(图5);IL-26鉴别诊断精神分裂症与痴呆的AUC为0.81,p=0.002344,特异性0.79,灵敏度0.95(图6)。
通过实验结果可知,IL-26对焦虑抑郁或精神分裂具有很好的诊断效果,且分别能在焦虑抑郁、精神分裂症和痴呆的两两之间进行鉴别诊断。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (8)
1.一种焦虑抑郁症或精神分裂症的诊断试剂在制备诊断焦虑抑郁症或精神分裂症产品中的应用,所述试剂包括检测IL-26表达水平的物质。
2.如权利要求1所述的应用,其特征在于,与正常组相比,所述IL-26在焦虑抑郁患者血清中显著降低,在精神分裂患者中显著降低,但明显低于焦虑抑郁患者。
3.如权利要求2所述的应用,其特征在于,所述试剂选自:识别IL-26的探针;或扩增IL-26引物;或结合IL-26蛋白的抗体。
4.检测IL-26表达水平的试剂在制备区分至少两种精神神经疾病的诊断产品中的应用,所述精神神经疾病包括焦虑抑郁症、精神分裂症和痴呆。
5.如权利要求4所述的应用,其特征在于,与正常组相比,所述IL-26在焦虑抑郁患者血清中显著降低,在精神分裂患者中显著降低,但明显低于焦虑抑郁患者,在痴呆患者中无显著差异;与痴呆患者相比,所述IL-26在焦虑抑郁患者中显著降低。
6.如权利要求5所述的应用,其特征在于,所述试剂选自:识别IL-26的探针;或扩增IL-26引物;或结合IL-26蛋白的抗体。
7.根据权利要求6所述的应用,其特征在于,所述产品包括制剂、试纸、芯片、试剂盒或系统。
8.根据权利要求7所述的应用,其特征在于,所述芯片包括蛋白芯片;所述试剂盒包括胶体金试剂盒和ELISA试剂盒。
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