摘要
目的:基于全外显子组测序和生物信息学分析,筛选早发高血压合并血钾代谢异常病人的易感基因,并探讨其潜在分子机制。方法:纳入57例早发高血压病人,根据入院时血钾水平将病人分为正常血钾组(血钾3.5~5.5 mmol/L,21例)和低血钾组(血钾<3.5mmol/L,36例)。采集外周血样本进行全外显子组测序及变异注释,筛选两组间交互性差异基因。采用DAVID平台进行基因本体(GO)功能富集、京都基因与基因组百科全书(KEGG)通路富集分析。利用STRING数据库构建蛋白-蛋白互作(PPI)网络,并导入Cytoscape软件进行拓扑分析,识别关键节点基因和挖掘功能模块。通过酶联免疫吸附试验(ELISA)检测两组血浆Ⅳ型胶原α3链(COL4A3)及Ⅳ型胶原α5链(COL4A5)蛋白表达水平。结果:两组年龄、左室舒张早期快速充盈的充盈峰流速(E峰)、左室舒张早期快速充盈的充盈峰流速/左室舒张晚期充盈的充盈峰流速(A峰)比值(E/A)、血钾、血钙水平及脑卒中/脑梗死比例比较,差异均有统计学意义(P<0.05)。基因筛查共识别出交互性差异基因63个。PPI网络拓扑分析显示,纤连蛋白1(FN1)、钾电压门控通道亚家族Q成员1(KCNQ1)、肌联蛋白(TTN)、神经纤维瘤病1型(NF1)、赖氨酸甲基转移酶2D(KMT2D)、胰岛素样生长因子2(IGF2)、Barttin蛋白(BSND)、钙电压门控通道亚家族α1 H亚基(CACNA1H)、钠通道上皮1型调节亚基γ(SCNN1G)、COL4A3及COL4A5等基因有较高的介数中心性(BC)。分子复合物检测(MCODE)聚类分析发现,FN1与COL4A3、Ⅳ型胶原α4链(COL4A4)、COL4A5及Ⅴ型胶原α1链(COL5A1)构成紧密连接的功能模块,显著富集于整合素信号通路。低血钾组血浆COL4A3和COL4A5水平均低于正常血钾组(P<0.05)。结论:通过外显子组测序和生物信息学整合分析筛选出FN1、KMT2D、NF1、TTN、COL4A3及COL4A5等早发高血压合并血钾代谢异常的潜在易感基因;其中,COL4A3与COL4A5可能是通过整合素信号通路参与疾病发生发展的关键分子,为阐明早发高血压的遗传机制提供了新的理论依据。
Objective:To screen the susceptible genes of patients with early-onset hypertension combined with abnormal potassium metabolism based on whole-exome sequencing and bioinformatics analysis,and to explore the potential molecular mechanism.Methods:A total of 57 patients with early-onset hypertension were included.According to the blood potassium at the time of admission,the patients were divided into the normal potassium group(blood potassium 3.5-5.5 mmol/L,21 cases)and the low potassium group(blood potassium<3.5 mmol/L,36 cases).Peripheral blood samples were collected for whole exome sequencing and variant annotation,and the genes with interactive differences between the two groups were screened.Gene ontology(GO)functional enrichment and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed using the DAVID platform.The protein-protein interaction(PPI)network was constructed using the STRING database and imported into Cytoscape software for topological analysis to identify key node genes and functional modules.The protein expressions of plasma typeⅣcollagenα3 chain(COL4A3)and typeⅣcollagenα5 chain(COL4A5)in the two groups were detected by enzyme-linked immunosorbent assay(ELISA).Results:There were statistically differences in term of age,early diastolic rapid filling velocity of the left ventricle(E peak),the ratio of early diastolic rapid filling velocity of the left ventricle to late diastolic filling velocity of the left ventricle(A peak)(E/A),blood potassium and blood calcium levels,and the proportion of stroke/cerebral infarction between the two groups(P<0.05).A total of 63 interactive differential genes were identified through genetic screening.The PPI network topological analysis revealed that genes such as fibronectin 1(FN1),potassium voltage-gated channel subfamily Q member 1(KCNQ1),myosin(TTN),neurofibromatosis type 1(NF1),lysine methyltransferase 2D(KMT2D),insulin-like growth factor 2(IGF2),Barttin protein(BSND),calcium voltage-gated channel subfamilyα1 H subunit(CACNA1H),sodium channel epithelial type 1 regulatory subunitγ(SCNN1G),COL4A3 and COL4A5 showed higher betweenness centrality(BC).The molecular complex detection(MCODE)clustering analysis revealed that FN1 and COL4A3,typeⅣcollagenα4 chain(COL4A4),COL4A5,and typeⅤcollagenα1 chain(COL5A1)formed a functional module of tight junction,which was significantly enriched in the integrin signaling pathway.The plasma levels of COL4A3 and COL4A5 in the low blood potassium group decreased more than those in the normal blood potassium group(P<0.05).Conclusion:Through exome sequencing and bioinformatics integration analysis,potential susceptibility genes of earlyonset hypertension combined with abnormal potassium metabolism were screened out,including FN1,KMT2D,NF1,TTN,COL4A3,and COL4A5.COL4A3 and COL4A5 might be key molecules involved in the occurrence and development of the disease through the integrin signaling pathway.It might be theoretical basis of early-onset hypertension.
作者
张玉
高颖
沙吉旦·阿不都热衣木
迪丽胡玛尔·阿布来提
邢智
帕丽达·阿布来提
ZHANG Yu;GAO Ying;Shajidan·Abudureyimu;Dilihuma'er·Abulaiti;XING Zhi;Palida·Abulaiti(The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830000,Xinjiang,China)
出处
《中西医结合心脑血管病杂志》
2026年第6期821-828,共8页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
新疆维吾尔自治区“天山英才”培养计划(No.TSYC202401B097)。
