颅内动脉瘤患者血清hsa-miR-513b-5p水平及作用机制研究  

Serum hsa-miR-513b-5p expression level and its mechanism of action in patients with intracranial aneurysm

作　　者：陈艳[1,2] 郑峥[1,2] 程琼[1,2] 汪银洲[1,2] CHEN Yan;ZHENG Zheng;CHENG Qiong;WANG Yinzhou(Provincial Clinical Medical College of Fujian Medical University,Fuzhou 350001,China;Fujian Provincial Hospital,Fuzhou 350001,China)

机构地区：[1]福建医科大学省立临床医学院,福建福州350001 [2]福建省立医院,福建福州350001

出　　处：《中国实用神经疾病杂志》2021年第12期1026-1034,共9页Chinese Journal of Practical Nervous Diseases

基　　金：福建省自然科学基金卫生联合面上项目(编号:2017J01176,2017J01240);福建省卫生健康中青年骨干人才培养项目(编号:2019-ZQN-3)。

摘　　要：目的探讨血清hsa-miR-513b-5p在颅内动脉瘤患者中的表达水平及其可能的作用机制。方法收集颅内动脉瘤患者(颅内动脉瘤组)和无颅内动脉瘤的志愿者(对照组)各100例,采用定量逆转录PCR(quantitative reverse transcription PCR,qRT-PCR)技术测定血清hsa-miR-513b-5p、肿瘤坏死因子(TNF-α)、MMP2、MMP3、MMP9(基质金属蛋白酶,MMPs)和基质金属蛋白酶抑制剂4(TIMP4)水平。采用hsa-miR-513b-5p模拟物和抑制剂转染人血管平滑肌细胞(human aortic smooth muscle cells,HASMC),检测细胞增殖、凋亡和坏死变化。通过双荧光素酶报告基因分析hsa-miR-513b-5p的作用靶点COL1A1,并分析影响颅内动脉瘤形成的信号通路。结果颅内动脉瘤组患者高密度脂蛋白胆固醇(HDL-C)、TNF-α、MMP2、MMP3、MMP9水平高于对照组(P<0.05),而低密度脂蛋白胆固醇(LDL-C)、N末端B型利钠肽原(NT-proBNP)、血浆总同型半胱氨酸(Hcy)和TIMP4水平降低(P<0.05)。与对照组相比,颅内动脉瘤组患者血清hsa-miR-513b-5p表达水平降低,差异有统计学意义(P<0.05)。经has-miR-513b-5p模拟物转染的HASMC,与模拟物对照组比较,其细胞增殖能力降低,而凋亡和坏死水平增高,差异均有统计学意义(P<0.05)。经has-miR-513b-5p抑制剂转染的HASMC,与抑制剂对照组比较,其细胞增殖能力增强,而凋亡和坏死水平下降,差异均有统计学意义(P<0.05)。双荧光素酶检测结果显示has-miR-513b-5p可靶向抑制COL1A1表达,并上调RIP1-RIP3-MLKL和MMPs信号通路,抑制HASMC增殖和细胞外基质合成。结论血清hsa-miR-513b-5p在颅内动脉瘤患者中表达水平下降,并参与调节血管平滑肌细胞增殖和细胞外基质合成影响动脉瘤形成。Objective To explore the expression level of serum hsa-miR-513b-5p in patients with intracranial aneurysm and its possible mechanism.Methods The quantitative reverse transcription PCR(RT-qPCR)was used to measure hsa-miR-513b-5p,TNF-α,MMP2,MMP3,MMP9 and TIMP4 expression in intracranial aneurysm patients(n=100)and healthy controls(n=100).Human aortic smooth muscle cells(HASMC)were cultured and transfected with hsa-miR-513b-5p mimic and inhibitor.Then cell proliferation,apoptosis and death were detected.Next,the dual luciferase reporter assay was be used to analyze the targeted relationships of hsa-miR-513b-5p with COL1A1 and analyze the relation signal pathways that affect the formation of intracranial aneurysms.Results High density lipoprotein(HDL-C),TNF-α,MMP2,MMP3,and MMP9 in the intracranial aneurysm group were higher than those in the control group(P<0.05),while low density lipoprotein(LDL-C),N-terminal pro-B-type natriuretic peptide(NT-proBNP),total homocysteine(Hcy)in plasma and TIMP4 were reduced(P<0.05).Compared with the control group,the serum hsa-miR-513b-5p expression level in the intracranial aneurysm group was decreased and the difference was statistically significant(P<0.05).The proliferation ability of HASMC transfected with miR-513b-5p mimic was decreased than mimic NC group,while cell apoptosis and death were increased,and there was a statistically significant between the two groups(P<0.05).The HASMC proliferation ability was enhance in miR-513b-5p inhibitor group than inhibitor NC group,while cell apoptosis and death were decreased,and there was a statistically significant between the two groups(P<0.05).Luciferase reporter assay results were confirmed that the COL1A1 was the direct target of miR-513b-5p,miR-513b-5p inhibited HASMC proliferation and extracellular matrix synthesis via up-regulating the RIP1-RIP3-MLKL and MMPs signaling pathways.Conclusion The expression of hsa-miR-513b-5p was significantly decreased in intracranial aneurysm patients,and it was involved in regulating HASMC prolife

关 键 词：颅内动脉瘤 hsa-miR-513b-5p COL1A1 RIP1-RIP3-MLKL通路 MMPs通路 细胞增殖 细胞外基质合成 

分 类 号：R739.41[医药卫生—肿瘤]