miR-98通过调控ox-LDL诱导血管内皮损伤抑制动脉粥样硬化形成的作用机制研究
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摘要
目的探讨miR-98对动脉粥样硬化(AS)发生、发展的调控作用及可能的作用机制。方法比较AS患者和健康受试者血浆微小核糖核酸(miR-98)和凝集素样氧化低密度脂蛋白受体-1(LOX-1)表达情况,采用MTT法检测氧化低密度脂蛋白(ox-LDL)干预对人主动脉内皮细胞(HAEC)、人主动脉平滑肌细胞(HASMC)和小鼠腹腔巨噬细胞增殖活性的影响,采用RT-PCR法检测ox-LDL干预对miR-98表达的影响,采用RT-PCR和Western blot法检测miR-98模拟物或抑制物转染细胞对LOX-1、细胞核因子κB(NF-κB p65)和内皮素-1(ET-1)表达的影响。结果与健康受试者比较,AS患者血浆miR-98表达下调,LOX-1表达上调(P<0.05)。在ox-LDL 15 ug/ml干预后,HAEC细胞和巨噬细胞增殖活性最强(P<0.05),HAEC和巨噬细胞中miR-98表达量较AS组明显降低(P<0.05)。与对照组和miR-98抑制物转染细胞比较,miR-98过表达可明显抑制LOX-1、NF-κB p65和ET-1的表达量(P<0.05)。结论 miR-98可能通过负性调控NF-κB信号通路抑制ox-LDL诱导的动脉粥样硬化早期血管内皮损伤过程。
Objective To investigate the regulation effect of miR-98 on occurrence and development of atherosclerosis(AS) and possible effective mechanism. Methods The expressions of plasma miR-98 and lectinlike oxidized low-density lipoprotein receptor-1(LOX-1) were compared between AS patients(AS group) and healthy controls(control group). The influence of oxidized low density lipoprotein(ox-LDL) on proliferative activities of HAEC, HASMC and macrophage was detected by using MTT. The influence of ox-LDL intervention on expression of miR-98 was detected by using RT-PCR. The influence of miR-98 simulant transfection cells or inhibitor transfection cells on expressions of LOX-1, nuclear factor-κB(NF-κB) and endothelin-1(ET-1) in were detected by using RT-PCR and Western blotting assay. Results The expression of miR-98 decreased and LOX-1 expression increased in AS group compared with control group(P<0.05). After ox-LDL(15 ug/m L) intervention, proliferative activities of HAEC and macrophage were the highest(P<0.05), and expression of miR-98 in HAEC and macrophage decreased significantly compared with AS group(P<0.05). The over-expression of miR-98 imulant transfection cells inhibited significantly expressions of LOX-1, NF-κB p65 and ET-1 compared with miR-98 inhibitor transfection cells(P<0.05). Conclusion Mi R-98 can relieve vascular endothelial injury induced by negatively regulating NF-κB signal pathway to inhibit ox-LDL in early AS stage.
Objective To investigate the regulation effect of miR-98 on occurrence and development of atherosclerosis(AS) and possible effective mechanism. Methods The expressions of plasma miR-98 and lectinlike oxidized low-density lipoprotein receptor-1(LOX-1) were compared between AS patients(AS group) and healthy controls(control group). The influence of oxidized low density lipoprotein(ox-LDL) on proliferative activities of HAEC, HASMC and macrophage was detected by using MTT. The influence of ox-LDL intervention on expression of miR-98 was detected by using RT-PCR. The influence of miR-98 simulant transfection cells or inhibitor transfection cells on expressions of LOX-1, nuclear factor-κB(NF-κB) and endothelin-1(ET-1) in were detected by using RT-PCR and Western blotting assay. Results The expression of miR-98 decreased and LOX-1 expression increased in AS group compared with control group(P<0.05). After ox-LDL(15 ug/m L) intervention, proliferative activities of HAEC and macrophage were the highest(P<0.05), and expression of miR-98 in HAEC and macrophage decreased significantly compared with AS group(P<0.05). The over-expression of miR-98 imulant transfection cells inhibited significantly expressions of LOX-1, NF-κB p65 and ET-1 compared with miR-98 inhibitor transfection cells(P<0.05). Conclusion Mi R-98 can relieve vascular endothelial injury induced by negatively regulating NF-κB signal pathway to inhibit ox-LDL in early AS stage.
引文
[1]董慧,于波.动脉粥样硬化斑块组成和治疗的新进展[J].中国循证心血管医学,2015,7(10):713-5.
[2]任广理,罗涛,侯伯轩,等.血浆炎症细胞因子与急性冠脉综合征的相关性研究[J].中国循证心血管医学,2017,9(3):360-4.
[3]Mark W,Feinberg,Kathryn J,et al.Micro RNA regulation of atherosclerosis[J].Circ Res,2016,118(4):703-20.
[4]Wuyuan Zhou,Benkui Zou,Lisheng Liu,et al.Micro RNA-98 acts as a tumor suppressor in hepatocellular carcinoma via targeting SALL4[J].Oncotarget,2016,7(45):74059-73.
[5]翟晓斌,葛朝明,范祯祯,等.急性缺血性卒中患者血清Ox-LDL水平与疾病严重程度及预后的相关性[J].中国循证心血管医学,2016,8(4):436-8.
[6]Zhibo Chen,Mian Wang,Qiong He,et al.Micro RNA-98 rescues proliferation and alleviates ox-LDL-induced apoptosis in HUVECs by targeting LOX-1[J].Exp Ther Med,2017,13(5):1702-10.
[7]Barbara Rizzacasa,Elena Morini,Sabina Pucci,et al.LOX-1 and its splice variants:a new challenge for atherosclerosisandcancertargeted therapies[J].Int J Mol Sci,2017,18(2):290.
[8]Giuseppe Danilo Norata,Federica Sala,Alberico Luigi Catapano,et al.Micro RNAs and lipoproteins:a connection beyond atherosclerosis?[J].Atherosclerosis,2013,227(2):209-15.
[9]Viola Neudecker,Kelley S Brodsky,Simone Kreth,et al.Emerging roles for micro RNAs in perioperative medicine[J].Anesthesiology,2016,124(2):489-506.
[10]Petra Hartmann,Andreas Schober,Christian Weber.Chemokines and micro RNAs in atherosclerosis[J].Cell Mol Life Sci,2015,72(17):3253-66.
[11]Ying-Ying Jin,Qing-Juan Chen,Yang Wei,et al.Upregulation of micro RNA-98 increases radiosensitivity in esophageal squamous cell carcinoma[J].J Radiat Res,2016,57(5):468-476.
[12]Yan-De Zhou,Xue-Qin Cao,Zhi-Hua Liu,et al.Rapamycin Inhibits Oxidized Low Density Lipoprotein Uptake in Human Umbilical Vein Endothelial Cells via m TOR/NF-κB/LOX-1 Pathway[J].PLo S One,2016,11(1):e0146777.
[13]Tiewei Lu,Sampath Parthasarathy,Hong Hao,et al.Reactive Oxygen Species Mediate Oxidized Low-Density Lipoprotein-Induced Inhibition of Oct-4 Expression and Endothelial Differentiation of Bone Marrow Stem Cells[J].Antioxid Redox Signal,2010,13(12):1845-56.
[14]陆荣荣,呼小龙,吾麦尔江·克力木,等.高尿酸血症对冠状动脉粥样硬化性心脏病大鼠炎性指标及氧化型低密度脂蛋白与凝集素样氧化型低密度脂蛋白受体1水平的影响[J].中国医药,2017,12(10):1487-90.
[15]Mingxin Liu,Guizhou Tao,Qifeng Liu,et al.Micro RNA let-7g alleviates atherosclerosis via the targeting of LOX-1 in vitro and in vivo[J].Int J Mol Med,2017,40(1):57-64.
[16]Yefei Zhang,Naiyun Chen,Jihao Zhang,et al.Hsa-Let-7g mi RNA Targets Caspase-3 and Inhibits the Apoptosis Induced by ox-LDL in Endothelial Cells[J].Int J Mol Sci,2013,14(11):22708-20.
[2]任广理,罗涛,侯伯轩,等.血浆炎症细胞因子与急性冠脉综合征的相关性研究[J].中国循证心血管医学,2017,9(3):360-4.
[3]Mark W,Feinberg,Kathryn J,et al.Micro RNA regulation of atherosclerosis[J].Circ Res,2016,118(4):703-20.
[4]Wuyuan Zhou,Benkui Zou,Lisheng Liu,et al.Micro RNA-98 acts as a tumor suppressor in hepatocellular carcinoma via targeting SALL4[J].Oncotarget,2016,7(45):74059-73.
[5]翟晓斌,葛朝明,范祯祯,等.急性缺血性卒中患者血清Ox-LDL水平与疾病严重程度及预后的相关性[J].中国循证心血管医学,2016,8(4):436-8.
[6]Zhibo Chen,Mian Wang,Qiong He,et al.Micro RNA-98 rescues proliferation and alleviates ox-LDL-induced apoptosis in HUVECs by targeting LOX-1[J].Exp Ther Med,2017,13(5):1702-10.
[7]Barbara Rizzacasa,Elena Morini,Sabina Pucci,et al.LOX-1 and its splice variants:a new challenge for atherosclerosisandcancertargeted therapies[J].Int J Mol Sci,2017,18(2):290.
[8]Giuseppe Danilo Norata,Federica Sala,Alberico Luigi Catapano,et al.Micro RNAs and lipoproteins:a connection beyond atherosclerosis?[J].Atherosclerosis,2013,227(2):209-15.
[9]Viola Neudecker,Kelley S Brodsky,Simone Kreth,et al.Emerging roles for micro RNAs in perioperative medicine[J].Anesthesiology,2016,124(2):489-506.
[10]Petra Hartmann,Andreas Schober,Christian Weber.Chemokines and micro RNAs in atherosclerosis[J].Cell Mol Life Sci,2015,72(17):3253-66.
[11]Ying-Ying Jin,Qing-Juan Chen,Yang Wei,et al.Upregulation of micro RNA-98 increases radiosensitivity in esophageal squamous cell carcinoma[J].J Radiat Res,2016,57(5):468-476.
[12]Yan-De Zhou,Xue-Qin Cao,Zhi-Hua Liu,et al.Rapamycin Inhibits Oxidized Low Density Lipoprotein Uptake in Human Umbilical Vein Endothelial Cells via m TOR/NF-κB/LOX-1 Pathway[J].PLo S One,2016,11(1):e0146777.
[13]Tiewei Lu,Sampath Parthasarathy,Hong Hao,et al.Reactive Oxygen Species Mediate Oxidized Low-Density Lipoprotein-Induced Inhibition of Oct-4 Expression and Endothelial Differentiation of Bone Marrow Stem Cells[J].Antioxid Redox Signal,2010,13(12):1845-56.
[14]陆荣荣,呼小龙,吾麦尔江·克力木,等.高尿酸血症对冠状动脉粥样硬化性心脏病大鼠炎性指标及氧化型低密度脂蛋白与凝集素样氧化型低密度脂蛋白受体1水平的影响[J].中国医药,2017,12(10):1487-90.
[15]Mingxin Liu,Guizhou Tao,Qifeng Liu,et al.Micro RNA let-7g alleviates atherosclerosis via the targeting of LOX-1 in vitro and in vivo[J].Int J Mol Med,2017,40(1):57-64.
[16]Yefei Zhang,Naiyun Chen,Jihao Zhang,et al.Hsa-Let-7g mi RNA Targets Caspase-3 and Inhibits the Apoptosis Induced by ox-LDL in Endothelial Cells[J].Int J Mol Sci,2013,14(11):22708-20.