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为了考察microRNA-150 (miR-150)对深静脉血栓形成(DVT)大鼠模型炎症反应的调控作用,本研究建立了DVT大鼠模型,通过RT-PCR检测miR-150的表达,并对DVT大鼠注射miR-150过表达慢病毒或阴性对照慢病毒。利用ELISA试剂盒检测大鼠血清纤溶酶原激活物抑制剂(PAI-1)、肿瘤坏死因子α(TNF-α)、白介素6 (IL-6)和白介素8 (IL-8)水平;并检测大鼠的血小板聚集抑制率和血栓烷B2 (TXB2)含量,通过苏木精伊红(HE)染色评价血栓形成,应用miR-150模拟物、miR-150抑制剂和阴性对照转染大鼠血管内皮细胞。Western blotting检测了大鼠血管内皮细胞中NF-κB p50的表达且使用CCK-8法和流式细胞仪检测了细胞活力和细胞凋亡。研究结果显示,DVT大鼠的miR-150表达水平显著低于正常大鼠(p<0.05)。miR-150的过表达降低了DVT大鼠的血小板聚集抑制率、TXB2含量和血管阻塞程度评分(p<0.05)。miR-150的过表达显著降低DVT大鼠和血管内皮细胞中PAI-1、TNF-α、IL-6和IL-8的水平(p<0.05)。miR-150的过表达促进了血管内皮细胞的增殖,而抑制了细胞凋亡(p<0.05)。miR-150的过表达下调了血管内皮细胞NF-κB p50的表达水平(p<0.05)。本研究表明,miR-150的上调可通过抑制NF-κB p50信号通路来抑制血栓形成和炎症反应。
Abstract：To investigate the regulatory effect of microRNA-150(miR-150) on the inflammatory response in deep vein thrombosis(DVT) rat models.In this study,a DVT rat model was established,and the expression of miR-150 was detected by RT-PCR.Then,DVT rats were injected with miR-150 overexpression lentivirus or negative control lentivirus.Serum plasminogen activator inhibitor(PAI-1),tumor necrosis factor alpha(TNF-α),interleukin 6(IL-6),and interleukin 8(IL-8) were detected by ELISA kit.The inhibition rate of platelet aggregation and the content of thromboxane B2(TXB2) in rats were measured.Thrombosis was evaluated by hematoxylin and eosin(HE)staining.Rat vascular endothelial cells were transfected with miR-150 mimics,miR-150 inhibitors,and negative controls,respectively.The expression of NF-κB p50 in rat vascular endothelial cells was detected by Western blotting.Cell viability and apoptosis were measured using the CCK-8 method and flow cytometry.The results showed that miR-150 expression in DVT rats was significantly lower than that in normal rats(p <0.05).Overexpression of miR-150 reduced the platelet aggregation inhibition rate,TXB2 content,and vascular occlusion score in DVT rats(p <0.05).The overexpression of miR-150 significantly reduced the levels of PAI-1,TNF-α,IL-6 and IL-8 in DVT rats and vascular endothelial cells(p <0.05).MiR-150 overexpression promoted the proliferation of vascular endothelial cells and inhibited apoptosis(p <0.05).Overexpression of miR-150 down-regulated the expression of NF-κB p50 in vascular endothelial cells(p<0.05).This study indicates that up-regulation of miR-150 can inhibit thrombosis and inflammatory response by inhibiting the NF-κB p50 signaling pathway.
Bao C.X.,Zhang D.X.,Wang N.N.,Zhu X.K.,Zhao Q.,and Sun X.L.,2018,MicroRNA-335-5p suppresses lower extremity deep venous thrombosis by targeted inhibition of PAI-1 via the TLR4 signaling pathway,J.Cell.Biochem.,119(6):4692-4710
Diaz J.A.,Obi A.T.,Myers D.D.,Wrobleski S.K.,Henke P.K.,Mackman N.,and Wakefield T.W.,2012,Critical review of mouse models of venous thrombosis,Arterioscler.Thromb.Vasc.Biol.,32(3):556-562
Dull T.,Zufferey R.,Kelly M.,Mandel R.J.,Nguyen M.,Trono D.,and Naldini L.,1998,A third-generation lentivirus vector with a conditional packaging system,J.Virol.,72(11):8463-8471
Fuchs T.A.,Brill A.,and Wagner D.D.,2012,Neutrophil extracellular trap(NET)impact on deep vein thrombosis,Arterioscler.Thromb.Vasc.Biol.,32(8):1777-1783
Hugo T.C.,2016,MicroRNA and venous thrombosis,Thromb.Haemost.,116(2):205
Li Y.D.,Ye B.Q.,Zheng S.X.,Wang J.T.,Wang J.G.,Chen M.,Liu J.G.,Pei X.H.,Wang L.J.,Lin Z.X.,Gupta K.,Mackman N.,Slungaard A.,Key N.S.,and Geng J.G.,2009,NF-κB transcription factor p50 critically regulates tissue factor in deep vein thrombosis,J.Biol.Chem.,284(7):4473-4483
Liu N.,Sun Q.,Chen J.,Li J.,Zeng Y.,Zhai S.,Li P.,Wang B.,and Wang X.,2012a,MicroRNA-9 suppresses uveal melanoma cell migration and invasion through the NF-κB1pathway,Oncol.Rep.,28(3):961-968
Liu S.J.,Kumar S.M.,Lu H.Z.,Liu A.H.,Yang R.F.,Pushparajan A.,Guo W.,and Xu X.W.,2012b,MicroRNA-9 up-regulates E-cadherin through inhibition of NF-κB 1-Snail1 pathway in melanoma,J.Pathol.,226(1):61-72
Marchand A.,Proust C.,Morange P.E.,LompréA.M.,and Tr gou觕t D.A.,2012,mi R-421 and mi R-30c inhibit SERPINE1 gene expression in human endothelial cells,PLoS One,7(8):e44532
Mo J.W.,Zhang D.F.,and Yang R.Z.,2016,MicroRNA-195 regulates proliferation,migration,angiogenesis and autophagy of endothelial progenitor cells by targeting GABARAPL1,Biosci.Rep.,36(5):e00396
Pan Y.,Liang H.W.,Liu H.,Li D.H.,Chen X.,Li L.M.,Zhang C.Y.,and Zen K.,2014,Platelet-secreted microRNA-223 promotes endothelial cell apoptosis induced by advanced glycation end products via targeting the insulin-like growth factor1 receptor,J.Immunol.,192(1):437-446
Rajput C.,Tauseef M.,Farazuddin M.,Yazbeck P.,Amin M.R.,Avin B.V.,Sharma T.,and Mehta D.,2016,MicroRNA-150suppression of angiopoetin-2 generation and signaling is crucial for resolving vascular injury,Arterioscler.Thromb.Vasc.Biol.,36(2):380-388
Raskob G.E.,Silverstein R.,Bratzler D.W.,Heit J.A.,and White R.H.,2010,Surveillance for deep vein thrombosis and pulmonary embolism:recommendations from a national workshop,Am.J.Prev.Med.,38(4S):502-509
Sun X.H.,He S.L.,Wara A.K.M.,Icli B.,Shvartz E.,Tesmenitsky Y.,Belkin N.,Li D.,Blackwell T.S.,Sukhova G.K.,Croce K.,and Feinberg M.W.,2014,Systemic delivery of microRNA-181b inhibits nuclear factor-κB activation,vascular inflammation,and atherosclerosis in apolipoprotein E-deficient mice,Circ.Res.,114(1):32-40
Tay J.,Tiao J.,Hughes Q.,Jorritsma J.,Gilmore G.,and Baker R.,2017,Circulating microrna as thrombosis sentinels:caveats and considerations,Semin.Thromb.Hemost.,44(3):206-215
Thulin P.,Wei T.,Werngren O.,Cheung L.,Fisher R.M.,Grandér D.,Corcoran M.,and Ehrenborg E.,2013,MicroR-NA-9 regulates the expression of peroxisome proliferator-activated receptorδin human monocytes during the inflammatory response,Int.J.Mol.Med.,31(5):1003-1010
Weaver J.L.,Matheson P.J.,Hurt R.T.,Downard C.D.,McClain C.J.,Garrison R.N.,and Smith J.W.,2016,Direct peritoneal resuscitation alters hepatic mi RNA expression after hemorrhagic shock,J.Am.Coll.Surg.,223(1):68-75
Xie X.S.,Liu C.P.,Lin W.,Zhan B.M.,Dong C.J.,Song Z.,Wang S.L.,Qi Y.G.,Wang J.L.,and Gu Z.Q.,2016,Deep vein thrombosis is accurately predicted by comprehensive analysis of the levels of microRNA-96 and plasma D-dimer,Exp.Ther.Med.,12(3):1896-1900
Yu F.J.,Chen B.C.,Fan X.F.,Li G.J.,Dong P.H.,and Zheng J.J.,2017,Epigenetically-regulated microRNA-9-5p suppresses the activation of hepatic stellate cells via TGFBR1 and TGF-BR2,Cell.Physiol.Biochemi.,43(6):2242-2252
基本信息:
DOI：10.13417/j.gab.039.004753
中图分类号:R543.6;R-332
引用信息:
[1]田野,李新喜,白超,等.microRNA-150对深静脉血栓形成大鼠模型炎症反应的调控作用[J].基因组学与应用生物学,2020,39(10):4753-4760.DOI:10.13417/j.gab.039.004753.
2020-10-25
2020-10-25