MicroRNA-195参与切应力对血管内皮细胞炎症调控作用的研究  

Role of miR-195 in regulation of vascular endothelial cell inflammation by shear stress

作　　者：吴小苑[1] 范文冬 Xiao-yuan Wu;Wen-dong Fan(Guangdong Prevention and Treatment Center for Occupational Diseases,Guangzhou,Guangdong 510300,China;Department of Cardiology,the First Affiliated Hospital of Sun Yat-sen University,Guangzhou,Guangdong 510080,China)

机构地区：[1]广东省职业病防治院,广东广州510300 [2]中山大学附属第一医院心内科,广东广州510080

出　　处：《中国现代医学杂志》2018年第10期26-31,共6页China Journal of Modern Medicine

摘　　要：目的研究micro RNA-195(miR-195)是否参与切应力对血管内皮细胞(ECs)炎症的调控作用。方法 (1)验证生理切应力调节ECs miRNAs的表达:15 dyne/cm2的生理切应力处理人脐静脉内皮细胞(HUVEC),24 h后利用实时荧光定量聚合酶链反应(q RT-PCR)检测内皮细胞相关miRNAs(miR-27b、miR-143/145及miR-195等)的表达变化;(2)miR-195与内皮细胞炎症反应的关系:体外转染miR-195mimics及inhibitors至HUVEC,通过Western blot、免疫荧光检测ECs中炎症相关因子表达及miR-195对内皮细胞的炎症控制水平。结果 15 dyne/cm^2的生理切应力处理HUVEC,24 h后miR-27b、miR-143/145及miR-195等表达上调;过表达miR-195对ECs内皮型一氧化氮合酶(e NOS)的表达没有影响,但抑制了p65的磷酸化(P-p65)表达;采用miRNA inhibitors抑制内源性的miR-195后,ECs e NOS的表达下调,抑制了κB抑制蛋白(IκBα)的表达水平。免疫荧光结果显示,在ECs中过表达miR-195抑制了NF-κB的核转运,而抑制miR-195可能促进了NF-κB的核转运,促进炎症。结论生理切应力诱导miR-195表达上调,miR-195可能参与切应力对ECs炎症的调控作用。Objective To investigate whether miR-195 is involved in the regulation of vascular endothelial cell(EC)inflammation by shear stress.Methods To verify the regulation of EC miRNA expression by physiological shear stress,15 dyne/cm2 of physiological shear stress were used to process human umbilical vein endothelial cells(HUVECs),after 24 hours,the expressions of miRNAs(miR-27b,miR-143/145,miR-195,etc.)in endothelial cells were detected by qRT-PCR.To study the relationship between miR-195 and endothelial cell inflammatory response,miR-195 mimics and inhibitors were transfected into HUVECs in vitro,the expressions of inflammatory factors in ECs and the effect of miR-195 in controlling endothelial cell inflammation were detected using Western blot and immunofluorescence technique.Results After processing HUVECs with 15 dyne/cm2 of physiological shear stress for 24 hours,the expressions of miR-27b,miR-143/145 and miR-195 were significantly up-regulated(P<0.05).The over-expression of miR-195 did not affect the expression of eNOS,but significantly inhibited the expression of P-p65(P<0.05);while inhibition of endogenous miR-195 by miRNA inhibitors significantly reduced the expression of EC eNOS(P<0.05)and inhibited the expression of IKBa(P<0.05).Immunofluorescence results showed that overexpression of miR-195 in ECs inhibited nuclear translocation of NF-κB,while inhibition of miR-195 may promote the nuclear translocation of NF-κB and promote inflammation.Conclusions The expression of miR-195 is significantly up-regulated by physiological shear stress.miR-195 may be involved in the regulation of EC inflammation by shear stress.

关 键 词：miR-195 内皮细胞 炎症反应 

分 类 号：R54[医药卫生—心血管疾病]