Zhang Jianhua, Zhou Zheng, Zhang Ning, Jin Wenwen, Ren Yafeng, Chen Chuanliang
Medical Engineering Technology and Data Mining Institute of Zhengzhou University, No. 100 Science Ave., Gaoxin Dist., Zhengzhou 450001, China.
Department of Respiration, The Second Affiliated Hospital of Zhengzhou University, No. 2 Jingba Rd., Zhengzhou 450014, China.
Saudi J Biol Sci. 2017 Mar;24(3):582-588. doi: 10.1016/j.sjbs.2017.01.029. Epub 2017 Jan 26.
Our purpose was to investigate the regulatory mechanism of TRPV1 and related cytokines on children bronchial asthma. TRPV1 mRNA level and two SNP genotypes of children in case group and control group were detected by real-time quantitative PCR. Western blot and ELISA were used to measure the levels of cytokines like IgE, IL-2, etc. Their correlations were analyzed by Logistic regression and KEGG analysis. Moreover, tertiary structure of protein and miRNA binding sites were also predicted by online tools. Case group was obviously different from control group in TRPV1 mRNA level, the two SNP genotypes distribution and the related cytokines levels. Logistic regression analysis further demonstrated that TRPV1 mRNA level, EOS, IL-4 and IL-5 may be risk factors for children bronchial asthma. And based on that, the preliminary regulatory network of children bronchial asthma was drawn. What's more, mutation of rs4790521 and rs4790522 in TRPV1 gene both induced its corresponding miRNA binding site's change. The preliminary regulatory network of TRPV1 and related cytokines on children bronchial asthma established in this study provides certain theoretical basis for pathogenesis and treatment of children bronchial asthma.
我们的目的是研究瞬时受体电位香草酸亚型1（TRPV1）及相关细胞因子对儿童支气管哮喘的调控机制。采用实时定量聚合酶链反应（PCR）检测病例组和对照组儿童的TRPV1信使核糖核酸（mRNA）水平及两种单核苷酸多态性（SNP）基因型。采用蛋白质免疫印迹法和酶联免疫吸附测定法（ELISA）检测免疫球蛋白E（IgE）、白细胞介素-2（IL-2）等细胞因子水平。通过逻辑回归分析和京都基因与基因组百科全书（KEGG）分析对它们之间的相关性进行分析。此外，还利用在线工具预测了蛋白质的三级结构和微小核糖核酸（miRNA）结合位点。病例组与对照组在TRPV1 mRNA水平、两种SNP基因型分布及相关细胞因子水平上存在明显差异。逻辑回归分析进一步表明，TRPV1 mRNA水平、嗜酸性粒细胞（EOS）、IL-4和IL-5可能是儿童支气管哮喘的危险因素。在此基础上，绘制了儿童支气管哮喘的初步调控网络。此外，TRPV1基因中rs4790521和rs4790522的突变均导致其相应miRNA结合位点的改变。本研究建立的TRPV1及相关细胞因子对儿童支气管哮喘的初步调控网络为儿童支气管哮喘的发病机制及治疗提供了一定的理论依据。
