CN116970694A - miR-9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用 - Google Patents
miR-9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用 Download PDFInfo
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Abstract
本发明公开了miR‑9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用，属于分子检测技术领域。本发明提供了一种增加miR‑9和/或miR‑9类似物的含量或表达量的试剂在制备诊断和/或治疗细胞炎症的工具中的应用。本发明AFB1能够促进RelA和E3泛素连接酶TRIM7相互作用，增加转录因子RelA泛素化降解，继而下调miR‑9的表达，从而调控其对CXCR4的抑制作用。因此，恢复miR‑9的表达可以拮抗上游信号，阻断炎症反应的发生；或者设计并合成miR‑9的类似物，或通过载体表达miR‑9，同样可以抑制AFB1对肾小球足细胞的促炎作用。
Description
技术领域
本发明属于分子检测技术领域，具体涉及miR-9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用。
背景技术
microRNA(miRNA)是一种小的非编码RNA，能够对细胞生长、分化和发病机制相关的基因表达进行转录后调控。研究发现，microRNA-9(miR-9)靶向炎症趋化因子受体CXCR4，通过与CXCR4的C-X-C基序相互作用，介导相关信号通路，能够抑制高糖诱导的人脐静脉内皮损伤过程中的细胞凋亡。相反地，由链脲佐菌素诱导糖尿病的小鼠模型中，miR-9在肾小球足细胞表达水平会下降。这些发现表明了miR-9可能与肾小球足细胞损伤存在相关性。此外，据报道，CXCR4能提高硫氧还蛋白互作蛋白(TXNIP)和NOD样受体热蛋白结构域相关蛋白3(NLRP3)的表达水平，从而影响下游炎性因子的分泌。敲除CXCR4可降低TXNIP水平，体现TXNIP是CXCR4信号传导的下游效应蛋白。综上所述，CXCR4可能通过调控TXNIP/NLRP3炎性小体信号轴触发炎症反应。
黄曲霉毒素B1(AFB1)属于真菌次级代谢物，是最常见的环境致癌物，被国际癌症研究机构(IARC)列为人类I类致癌物，危害性极强。尽管AFB1经肝微粒体酶代谢，但残留部分则由肾脏排泄，表明AFB1必然会是诱导肾功能性障碍的重要原因之一。由此可见，明确AFB1诱导肾脏炎症的具体分子机制迫在眉睫。以前人的研究结果为基础，miR-9下调能否通过CXCR4/TXNIP/NLRP3炎性小体通路参与AFB1诱导的肾小球足细胞炎症，仍有待阐明。RelA(又称p65)是NF-κB的一个亚基，由p105前体和p52衍生的转录因子，它与NF-κB亚单位p50结合形成RelA-p50异源二聚体，用于核易位，并在代谢障碍、肿瘤发生、病毒感染等方面发挥重要作用。已有研究发现，RelA还参与调控miRNA的基因表达。但是，RelA是否参与了调控miR-9的表达，进而导致AFB1诱导的肾小球足细胞炎症，还有待继续探索。因此，尽早地研发出靶向性强，有效且合适的治疗药物及诊断试剂是目前面对AFB1诱导肾小球足细胞炎症的首要措施。
发明内容
本发明的目的在于提供miR-9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用，明确miR-9与趋化因子受体CXCR4的关系，并在体外和体内模型中发现miR-9的低表达与AFB1诱导的肾小球足细胞炎症密切相关。
本发明提供了一种增加miR-9和/或miR-9类似物的含量或表达量的试剂在制备诊断和/或治疗细胞炎症的工具中的应用。
优选的，所述增加miR-9和/或miR-9类似物的含量或表达量的试剂包括以下至少一种：恢复miR-9表达的试剂、miR-9类似物、包含miR-9的表达载体和miR-9激动剂。
优选的，所述miR-9类似物的引物包括核苷酸序列如SEQ ID No.45所示的上游引物和SEQ ID No.46所示的下游引物。
优选的，所述细胞炎症包括AFB1诱导的细胞炎症。
优选的，所述治疗包括以下至少一项：(1)拮抗AFB1诱导的RelA和E3泛素连接酶TRIM7的相互作用；
(2)沉默E3泛素连接酶TRIM7的表达；
(3)拮抗趋化因子受体CXCR4/TXNIP/NLRP3信号轴诱导细胞炎症；
(4)恢复或增加转录因子RelA的表达；
(5)抑制转录因子RelA泛素化降解；
(6)抑制CXCR4基因表达。
本发明还提供了一种诊断AFB1诱导细胞炎症的试剂，包括检测miR-9的表达量的试剂。
优选的，还包括检测RelA、TRIM7和CXCR4/TXNIP/NLRP3信号轴相关分子的表达量的试剂。
优选的，所述细胞炎症包括肾小球足细胞炎症。
优选的，所述检测miR-9的表达量包括检测基因表达量、mRNA表达量或蛋白质表达量。
本发明还提供了一组测定miR-9表达量的引物，包括核苷酸序列如SEQ IDNo.3所示的上游引物和SEQ ID No.4所示的下游引物。
有益效果：本发明提供了一种增加miR-9和/或miR-9类似物的含量或表达量的试剂在制备诊断和/或治疗细胞炎症的工具中的应用。本发明中，miR-9可以通过抑制其靶基因趋化因子受体CXCR4的表达，调控TXNIP/NLRP3信号轴诱导肾小球足细胞炎症；AFB1能够促进RelA和E3泛素连接酶TRIM7相互作用，增加转录因子RelA泛素化降解，继而下调miR-9的表达，从而调控其对CXCR4的抑制作用。因此，恢复miR-9的表达可以拮抗上游信号，阻断炎症反应的发生；或者设计并合成miR-9的类似物(mimics)，或通过载体表达miR-9，同样可以抑制AFB1对肾小球足细胞的促炎作用。
附图说明
图1为AFB1通过下调小鼠肾小球及MPC-5中miR-9表达量，以及影响miR-9与CXCR4的结合图；A：采用qRT-PCR方法分别检测0.75mg/kgAFB1处理第0、2、4周小鼠肾小球(n＝6)和5、10、20μMAFB1处理的MPC-5(n＝6)中miR-9的相对表达量统计图；B：小鼠、大鼠和人之间miR-9与CXCR4序列结合位点的序列比对图；C：pMIR-CXCR4结合位点和pMIR-CXCR4结合位点突变的荧光素酶报告质粒示意图，以及miR-9对CXCR4结合位点的可能靶点；
图2为AFB1通过下调miR-9、上调CXCR4、并激活TXNIP/NLRP3通路诱导MPC-5炎症反应的模式图；“*”与相应细胞对照组比较，p<0.05；“#”与AFB1和对照mimic细胞组比较，p<0.05；A：采用qRT-PCR方法检测mimics转染MPC-5(n＝6)后miR-9的相对表达量；B：通过Western blot方法检测mimics转染MPC-5(n＝6)后CXCR4的相对蛋白表达量；C：将miR-9类似物(mimic)或miR-9对照类似物瞬时转染至暴露于20μMAFB1的MPC-5(n＝6)中，采用Westernblot方法检测CXCR4、nephrin、podocin以及NLRP3、ASC、Cleaved-casp1和TXNIP的相对蛋白表达量；D：将miR-9类似物(mimic)或miR-9对照类似物瞬时转染至暴露于20μMAFB1的MPC-5(n＝6)中，采用qRT-PCR和ELISA方法检测细胞中IL-1β、IL-6和TNF-αmRNA相对表达量和蛋白含量；
图3为AFB1通过下调miR-9、上调CXCR4、激活TXNIP/NLRP3通路诱导小鼠肾小球足细胞炎症的模式图；“*”与相应的正常动物比较，p<0.05；“#”与AFB1和rAAv9-CAG-GFP对照组比较，p<0.05；A：采用qRT-PCR检测CAG-GFP-miR-9或CAG-GFP荧光素酶对照载体转染0.75mg/kgAFB1处理的小鼠肾小球后miR-9的表达量；B：免疫荧光标记转染荧光素酶载体后的小鼠肾小球(n＝3)用GFP(绿色；标尺为25μm)和DAPI(蓝色；标尺为25μm)；C：Westernblot检测小鼠肾小球(n＝6)中CXCR4、nephrin、podocin以及NLRP3、ASC、Cleaved-casp1和TXNIP的相对蛋白表达量；D：通过ELISA法检测小鼠肾小球中IL-1β、IL-6和TNF-α的蛋白含量(n＝6)；E：HE染色在攻毒第4周时小鼠肾小球内炎性细胞浸润(n＝3，标尺为20μm)，箭头表示小鼠肾小球内炎性细胞因子浸润和系膜增厚；F：流式细胞术检测第4周时各实验组小鼠肾小球(n＝5)中表达IFN-γ和Gr-1的中性粒细胞比例和表达CD68和CD86的巨噬细胞比例；
图4为AFB1通过促进RelA泛素依赖性蛋白水解降低RelA并下调miR-9的表达模式图，LaminA作为RelA的内参蛋白，“*”与相应细胞对照组比较，p<0.05；“#”与AFB1和LV5-GFP细胞组比较，p<0.05；A：通过Westernblot检测0.75mg/kgAFB1处理第0、2、4周小鼠肾小球和5、10、20μMAFB1处理的MPC-5中RelA的相对蛋白表达量(n＝6)；B：采用qRT-PCR检测转染RelA siRNA或对照试剂的MPC-5中RelA调控的miRNA(n＝6)以及转染LV5-RelA的MPC-5中miR-9的相对表达量(n＝6)；C：ChIP-PCR检测暴露或未暴露AFB1及存在或不存在MG-132的MPC-5中miR-9启动子与RelA的结合情况；D：IP-MS检测与RelA相互作用的蛋白；E：通过IP结合Westernblot检测RelA与TRIM7及泛素的相互作用；F：通过IP结合Westernblot检测转染TRIM7 siRNA的MPC-5中RelA及其泛素化水平。
具体实施方式
本发明提供了一种增加miR-9和/或miR-9类似物的含量或表达量的试剂在制备诊断和/或治疗细胞炎症的工具中的应用。
本发明所述增加miR-9和/或miR-9类似物的含量或表达量的试剂优选包括以下至少一种：恢复miR-9表达的试剂、miR-9类似物、包含miR-9的表达载体和miR-9激动剂。本发明所称恢复miR-9表达的试剂优选包括表达RelA(GeneID:19697)进而恢复内源miR-9的表达如LV5-RelA和TRIM7 siRNA；所述miR-9类似物的引物包括核苷酸序列如SEQ ID No.45所示的上游引物(F：UCUUUGGUUAUCUAGCUGUAUGA)和SEQ ID No.46所示的下游引物(R：AUACAGCUAGAUAACCAAAGAUU)；包含miR-9的表达载体优选包括CAG-GFP-miR-9；miR-9激动剂优选包括在AFB1诱导的肾小球炎症中表达miR-9，或通过干预miR-9的上游信号。本发明所述miR-9(mmu-miR-9-5pMIMAT0000142)的核苷酸序列如SEQ ID No.48所示：UCUUUGGUUAUCUAGCUGUAUGA。
本发明所述细胞炎症优选包括AFB1诱导的细胞炎症，AFB1通过促进RelA和E3泛素连接酶TRIM7(tripartite motifcontaining 7)相互作用，增加转录因子RelA泛素化降解，继而下调miR-9的表达，刺激趋化因子受体CXCR4/TXNIP/NLRP3信号轴诱导肾小球足细胞炎症。因此，本发明所述治疗优选包括以下至少一项：(1)拮抗AFB1诱导的RelA和E3泛素连接酶TRIM7的相互作用；
(2)沉默E3泛素连接酶TRIM7的表达；
(3)拮抗趋化因子受体CXCR4/TXNIP/NLRP3信号轴诱导细胞炎症；
(4)恢复或增加转录因子RelA的表达；
(5)抑制转录因子RelA泛素化降解；
(6)抑制CXCR4基因表达。
本发明所述诊断优选包括检测miR-9及其相关联的RelA、TRIM7和CXCR4/TXNIP/NLRP3信号轴相关分子的表达，可以对AFB1诱导肾小球足细胞炎症进行早期诊断，对肾小球炎症的疾病发展状况和预后做出评价。
本发明还提供了一种诊断AFB1诱导细胞炎症的试剂，包括检测miR-9的表达量的试剂。
本发明优选还包括检测RelA、TRIM7和CXCR4/TXNIP/NLRP3信号轴相关分子的表达量的试剂。本发明所述检测优选包括检测基因表达量、mRNA表达量或蛋白质表达量。本发明对所述检测的方法并没有特殊限定，优选包括qRT-PCR、Westernblot或ELISA。
本发明还提供了一组测定miR-9表达量的引物，包括核苷酸序列如SEQ IDNo.3所示的上游引物和SEQ ID No.4所示的下游引物。
本发明所述上游引物的序列为：CGGGCTCTTTGGTTATCTAGC，下游引物的序列为：CAGCCACAAAAGAGCACAAT，退火温度为60℃，扩增产物优选为65bp。本发明所述引物中优选还包括用于反转录的茎环序列，所述茎环序列的核苷酸序列如SEQ ID No.47所示：CCTGTTGTCTCCAGCCACAAAAGAGCACAATATTTCAGGAGACAACAGG。
为了进一步说明本发明，下面结合实施例对本发明提供的miR-9及其类似物在制备诊断和/或治疗细胞炎症的药物中的应用进行详细地描述，但不能将它们理解为对本发明保护范围的限定。
实施例1
1、AFB1通过抑制小鼠肾小球及MPC-5中miR-9上调CXCR4
采用qRT-PCR方法分别检测0.75mg/kgAFB1处理第0、2、4周小鼠肾小球(n＝6)和5、10、20μMAFB1处理的MPC-5(n＝6)中miR-9的相对表达量(U6 RNA作为miRNA的内参参照)。
表1引物序列
结果如图1所示，经AFB1处理后，无论是小鼠还是细胞内，miR-9的表达量均显著降低(图1中A)。根据TargetScan 5.2(http://www.targetscan.org)发现，CXCR4是miR-9的靶基因之一，miR-9通过结合CXCR4基因的3’-UTR而发挥功能(图1中B)。
为了验证miR-9与CXCR4基因的结合，将野生型和突变型3’-UTR靶向序列构建到荧光素酶报告载体pMIR中，随后将miR-9和对照miRNA(引物序列SEQ ID No.7和SEQ ID No.8)与野生型或突变型荧光素酶报告载体(pMIR-REPORT Luciferase载体，将靶位点ACCAAAG突变为ACGGCGC)共转染到MPC-5细胞，结果如图1中C所示，miR-9同野生型CXCR43’-UTR的靶向结合使荧光值相对降低；相反，miR-9无法与突变型CXCR43’-UTR结合而使荧光值未改变，表明miR-9能够通过3’-UTR结合位点抑制CXCR4基因表达。而对照miRNA由于无法与野生型CXCR43’-UTR或突变型CXCR43’-UTR序列结合，荧光值皆未改变。
2、AFB1通过下调miR-9上调CXCR4激活TXNIP/NLRP3通路诱导MPC-5炎症反应
相关试剂包括：β-actin抗体(Abcam)、nephrin抗体(Abcam)、podocin抗体(Abcam)、CXCR4抗体(Abcam)、ASC抗体(Abcam)、NLRP3抗体(Abcam)、Pro-caspase1抗体(CST)、Cleaved-caspase 1抗体(CST)、TXNIP抗体(Abcam)、山羊抗兔IgG H&L(HRP)(CST)，ELISA试剂盒(R&D Systems Inc.)。
为了验证miR-9在AFB1处理的MPC-5细胞中调控CXCR4的作用。将miR-9mimic(引物为SEQ ID No.45和SEQ ID No.46)瞬时转染MPC-5细胞24小时，通过qRT-PCR检测miR-9mimic的转染效率，结果如图2所示，miR-9的表达显著增加(图2中A)，CXCR4的相对蛋白表达量显著降低(图2中B)。
将miR-9mimic和AFB1(24h)共处理的MPC-5细胞同miR-9对照类似物和AFB1共处理组相比，nephrin和podocin的相对蛋白表达量显著增加，CXCR4、TXNIP、NLRP3、ASC和Cleaved-casp1显著减少(图2中C)。miR-9mimic转染还能够导致AFB1处理的足细胞中IL-1β、IL-6和TNF-αmRNA相对表达量和蛋白含量减少(图2中D)。结果进一步证明了miR-9在AFB1诱导的小鼠肾小球足细胞炎症发生过程中的负调控作用。
表2引物序列
3、AFB1通过下调miR-9、增加CXCR4激活TXNIP/NLRP3通路诱导肾小球足细胞炎症
相关试剂包括：荧光标记Gr-1抗体(CST)，荧光标记IFN-γ抗体(CST)或CD86抗体(荧光二抗：山羊抗兔IgG)(Abcam)和CD68抗体(荧光二抗：山羊抗兔IgG)(Abcam)，其余相同的试剂同上。
为了解miR-9在AFB1诱导的足细胞炎症过程中下调CXCR4/TXNIP/NLRP3炎症小体通路的能力，采用qRT-PCR检测CAG-GFP-miR-9或CAG-GFP荧光素酶对照载体转染0.75mg/kgAFB1处理4周的小鼠肾小球后miR-9的表达量。
结果如图3所示，采用rAAV9-CAG-GFP-miR-9转染了AFB1处理的小鼠肾小球后，miR-9的表达显著增加(图3中A和B)。miR-9的表达导致CXCR4蛋白表达水平显著减少，nephrin和podocin显著升高，同时抑制了AFB1处理的小鼠肾小球中TXNIP、NLRP3、ASC和Cleaved-casp1升高(图3中C)。
此外，miR-9的升高也减轻了AFB1诱导的肾小球炎症，体现在炎症细胞因子IL-1β、IL-6和TNF-α分泌减少(图3中D)，炎性细胞浸润减轻(图3中E和F)。
4、AFB1通过促进RelA泛素依赖性蛋白水解而降低RelA、下调miR-9的表达
相关试剂包括：RelA抗体(Abcam)、TRIM7抗体(Abcam)、LaminA抗体(Abcam)、Ubiquitin抗体(Abcam)，其余相同的试剂同上。
为了探讨RelA是否参与AFB1降低肾小球足细胞中miR-9的表达，通过Westernblot检测0.75mg/kgAFB1处理第0、2、4周小鼠肾小球和5、10、20μMAFB1处理的MPC-5中RelA的相对蛋白表达量。
结果如图4所示，AFB1处理导致小鼠肾小球和MPC-5细胞中p-RelA/RelA显著降低(图4中A)。
为了研究RelA与肾小球足细胞MPC-5中miR-9表达的关系，采用qRT-PCR检测转染RelAsiRNA或对照试剂的MPC-5细胞中RelA调控的miRNA以及转染LV5-RelA的MPC-5中miR-9的相对表达量。结果表明，过表达RelA能够抵消AFB1对miR-9的抑制作用(图4中B)，说明AFB1可能是通过下调RelA从而减少miR-9的表达。
之后，为了分析RelA和miR-9启动子区域的结合情况，采用ChIP-PCR方法检测暴露或未暴露AFB1及存在或不存在MG-132的MPC-5中miR-9启动子与RelA的结合情况，即将细胞用1％甲醛固定10分钟，收集细胞，细胞核超声处理，并与RelA抗体(CST)或对照IgG在4℃下孵育过夜，进行免疫沉淀，从沉淀的免疫复合物中洗脱DNA，用PCR扩增与RelA结合的miR-9启动子DNA片段，然后用核酸凝胶电泳进行相对定量。结果表明，无论加入或者不加入蛋白酶体抑制剂MG-132，AFB1都能减少MPC-5细胞中miR-9启动子区域的RelA结合(图4中C)，说明减少的RelA在AFB1诱导的miR-9下调中发挥作用。
对于RelA结合蛋白的分析，采用IP-MS检测与RelA相互作用的蛋白。在4℃与RelA抗体孵育过夜后，将MPC-5裂解物与蛋白A/G琼脂糖珠混合，然后进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析和考马斯蓝染色，以分离目标蛋白。然后，蛋白质样本被送往苏州Mass-elife生物技术有限公司(中国苏州)进行进一步分析。在液相色谱-质谱(LC-MS)分析中，蛋白质在乙腈和水的混合物中重新溶解，样本数据分析使用苏州Mass-elife生物技术有限公司(中国苏州)的超高效液相色谱系统耦合高分辨率质谱仪。使用Byonic2.13.17(Protein Metrics,California,USA)在uniport人类蛋白质数据库上检索MS原始文件，对鉴定出的蛋白进行丰度分析。结果表明，AFB1处理的MPC-5与对应的对照组相比，TRIM7在AFB1处理组中显著富集(图4中D)。
随后，通过Co-IP揭示了在暴露或未暴露AFB1及存在或不存在MG-132的MPC-5中RelA与TRIM7、泛素的相互作用。简而言之，在4℃12,000×g下超速离心20分钟获得MPC-5全细胞裂解液，随后与RelA抗体和蛋白A/G琼脂糖珠(Amersham Biosciences)在4℃下孵育过夜。之后，用裂解缓冲液洗涤免疫复合物，用钠盐样品缓冲液洗脱，进行WB分析。结果表明，AFB1促进了RelA和TRIM7的结合，从而增加了RelA的泛素化，下调RelA蛋白的表达(图4中E)。最后，通过IP结合Westernblot检测转染TRIM7 siRNA的暴露于AFB1的MPC-5细胞中RelA及其泛素化水平。结果表明，沉默TRIM7减少了MPC-5中RelA的泛素化水平，但恢复了AFB1处理后的RelA蛋白水平(图4中F)。
综上所述，AFB1处理通过增强RelA与TRIM7的结合从而增加泛素依赖性RelA降解，导致miR-9表达下调。
表3引物序列
总之，AFB1通过促进RelA和E3泛素连接酶TRIM7(tripartite motif containing7)相互作用，增加转录因子RelA泛素化降解，继而下调miR-9的表达；miR-9通过作用于趋化因子受体CXCR4/TXNIP/NLRP3信号轴诱导肾小球足细胞炎症。
5、miR-9及其相关分子的表达检测与AFB1诱导的肾小球足细胞炎症的诊断
应用Real-time RT-PCR检测miR-9的表达，应用Real-time RT-PCR或Westernblot检测RelA和CXCR4/TXNIP/NLRP3信号轴相关分子的表达，可以对AFB1的肾损伤作用进行诊断。
通过与正常组织细胞对照，miR-9的表达下调及其相互关联的RelA的低表达和CXCR4/TXNIP/NLRP3信号轴相关分子的高表达提示AFB1诱导肾小球足细胞产生炎症反应。
6、以RelA-miR-9-CXCR4/TXNIP/NLRP3信号通路为靶点的抑制AFB1诱导的肾小球足细胞炎症的分子或策略
设计并合成miR-9的类似物(mimics)，或者通过载体表达miR-9，应用病毒或非病毒方法介导其转移进入体外或体内模型，抑制AFB1对肾小球足细胞的促炎作用；沉默E3泛素连接酶TRIM7的表达，从而更有效影响炎症信号通路等。
尽管上述实施例对本发明做出了详尽的描述，但它仅仅是本发明一部分实施例，而不是全部实施例，人们还可以根据本实施例在不经创造性前提下获得其他实施例，这些实施例都属于本发明保护范围。
Claims (10)
1.一种增加miR-9和/或miR-9类似物的含量或表达量的试剂在制备诊断和/或治疗细胞炎症的工具中的应用。
2.根据权利要求1所述应用，其特征在于，所述增加miR-9和/或miR-9类似物的含量或表达量的试剂包括以下至少一种：恢复miR-9表达的试剂、miR-9类似物、包含miR-9的表达载体和miR-9激动剂。
3.根据权利要求1或2所述应用，其特征在于，所述miR-9类似物的引物包括核苷酸序列如SEQ ID No.45所示的上游引物和SEQ ID No.46所示的下游引物。
4.根据权利要求1所述应用，其特征在于，所述细胞炎症包括AFB1诱导的细胞炎症。
5.根据权利要求1或4所述应用，其特征在于，所述治疗包括以下至少一项：(1)拮抗AFB1诱导的RelA和E3泛素连接酶TRIM7的相互作用；
(2)沉默E3泛素连接酶TRIM7的表达；
(3)拮抗趋化因子受体CXCR4/TXNIP/NLRP3信号轴诱导细胞炎症；
(4)恢复或增加转录因子RelA的表达；
(5)抑制转录因子RelA泛素化降解；
(6)抑制CXCR4基因表达。
6.一种诊断AFB1诱导细胞炎症的试剂，其特征在于，包括检测miR-9的表达量的试剂。
7.根据权利要求6所述试剂，其特征在于，还包括检测RelA、TRIM7和CXCR4/TXNIP/NLRP3信号轴相关分子的表达量的试剂。
8.根据权利要求6或7所述试剂，其特征在于，所述细胞炎症包括肾小球足细胞炎症。
9.根据权利要求6或7所述试剂，其特征在于，所述检测miR-9的表达量包括检测基因表达量、mRNA表达量或蛋白质表达量。
10.一组测定miR-9表达量的引物，其特征在于，包括核苷酸序列如SEQ IDNo.3所示的上游引物和SEQ ID No.4所示的下游引物。
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