新疆哈萨克族原发性高血压患者炎症应答相关基因表达谱及分子标志物筛选

mRNA expression profiles and molecular biomarkers involved in inflammatory response in Kazakh patients with essential hypertension from Xinjiang

  • 摘要: 目的 筛选新疆哈萨克族原发性高血压(EH)患者外周血单个核细胞(PBMC)中与高血压炎症应答相关的mRNA,为该民族EH发病的免疫学机制研究、临床诊断和精准防治提供实验依据和分子标志物。方法 选择2018年4月-2019年5月在新疆石河子大学医学院第一附属医院心内科就诊的30例哈萨克族EH患者作为高血压组,并选取民族和年龄匹配的30例非高血压者作为非高血压对照组。受试者经测定血压、血糖和血脂水平后提取并纯化其PBMC总RNA,每组选择6例患者,采用长链非编码RNA(lncRNA)/mRNA V4.0基因芯片进行差异表达基因的分析;采用基因集合富集分析(GSEA)确定相关信号通路,采用基因本体(GO)和京都基因和基因组百科全书(KEGG)信号通路富集分析预测差异表达基因的功能。在两组中(n=30)采用实时定量聚合酶链反应(qRT-PCR)验证芯片和生物信息学分析结果中炎症应答相关基因的表达。结果 高血压组血压水平高于非高血压对照组收缩压(157.2±7.4)比(114.7±5.9)mm Hg,t=10.970,P<0.001;舒张压(97.0±6.9)比(73.7±6.9)mm Hg,t=5.887,P<0.001,而两组间的血糖和血脂水平差异无统计学意义(P>0.05)。与非高血压对照相比,哈萨克族EH患者PBMC中共有522个差异表达mRNA,其中表达上调的有411个,表达下调的有111个。GO功能注释结果显示,在表达上调的mRNA中,与炎症应答相关的生物学过程主要包括免疫应答调节、免疫应答激活及其信号转导等;与炎症应答相关的分子功能主要涉及抗原结合、免疫球蛋白受体结合、肽抗原结合等。GSEA和KEGG通路富集分析结果显示,与炎症相关的基因簇和信号通路主要包括黏着斑、细胞黏附分子、趋化因子信号通路、白细胞跨内皮迁移、细胞因子与其受体相互作用、B细胞受体信号通路。qRT-PCR结果显示上述炎症相关信号通路中的14个基因的表达与mRNA芯片结果相一致。结论 获得了新疆哈萨克族EH患者炎症应答相关基因表达谱,其可为哈萨克族EH发病预测诊断、病情进展和疗效评估提供分子标志物,同时可为该民族高血压的干预治疗提供新的分子靶标。

     

    Abstract: Objective To identify hypertensive inflammation-related mRNA in peripheral blood mononuclear cells(PBMC) of essential hypertension(EH) patients from Chinese Kazakh population in Xinjiang, to better reveal the immune mechanisms of EH in Kazakh and provide specific molecular biomarkers for clinical diagnosis and targeted-specific treatments. Methods Thirty Kazakh EH patients who visited the Cardiovascular Department of the First Affiliated Hospital of Xinjiang Shihezi University from April 2018 to May 2019 were enrolled as EH group, and 30 age-matched non-hypertensive Kazakh people were enrolled as control group. After measurement of blood pressure, serum lipids and blood glucose, total RNA was isolated from PBMC of both groups. Differentially expressed genes(DEGs) were identified by human long non-coding RNA(lncRNA)/mRNA V4.0 microarray in 6 randomly selected samples from each group. Signaling pathways related to these DEGs were identified by gene set enrichment analysis(GSEA). Gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) biological pathway analyses were performed to predict the functions of the DEGs. Reverse transcription quantitative real-time PCR(qRT-PCR) was used to validate the differential expression pattern of inflammation-related genes. Results The blood pressure were significantly higher in the EH group than control group systolic blood pressure(157.2±7.4) vs(114.7±5.9)mm Hg, t=10.970, P<0.001;diastolic blood pressure(97.0±6.9) vs(73.7±6.9)mm Hg, t=5.887, P<0.001, while there was no significant difference in the serum lipids and blood glucose between two groups. There were 522 mRNA significantly differentially expressed in PBMC of EH patients compared to controls, with 411 up-regulated and 111 down-regulated. GO enrichment analysis of up-regulated mRNA showed that the biological processes related to inflammatory response mainly included immune response regulation, immune response activation and their signal transduction, and the molecular functions related to inflammatory response mainly included antigen binding, immunoglobulin receptor binding, peptide antigen binding, etc. GSEA and KEGG pathway enrichment analysis indicated several positively enriched gene sets and pathways related to inflammatory response, such as focal adhesion, cell adhesion molecules, chemokine signaling pathway, leukocyte transendothelial migration, cytokine-cytokine receptor interaction, B cell receptor signaling pathway. The results of qRT-PCR showed that the expression of 14 genes in the above inflammation related signal pathway was consistent with that of microarray data. Conclusions Gene expression profile of hypertensive inflammation is identified in EH patients from Chinese Kazakh population in Xinjiang. This study not only provides specific molecular biomarkers for predicting and diagnosing progression of EH, but also provides targeted-specific treatments for EH of Chinese Kazakh population in Xinjiang.

     

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