miR-144-3p通过MAPK信号通路调节血管平滑肌细胞表型转换的机制研究  

Mechanism of miR-144-3p Regulating the Contractive Phenotype of Vascular Smooth Muscle Cells by MAPK Signaling Pathway

作　　者：冯笑抄 林瑷琪 韩翔[1,2] FENG Xiao-chao;LIN Ai-qi;HAN Xiang(Department of Neurology,Shanghai Fifth People's Hospital,Fudan University,Shanghai 200240,China;Department of Neurology,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属第五人民医院神经内科,上海200240 [2]复旦大学附属华山医院神经内科,上海200040

出　　处：《中国临床神经科学》2023年第4期365-372,共8页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金委面上项目(编号:8227052180)。

摘　　要：目的 探讨miR-144-3p是否参与颈动脉夹层的发生以及经MAPK信号通路调节血管平滑肌细胞表型转换的机制。方法 (1)将3周龄SD雄性大鼠随机分为AAV-NC组和AAV-miR-144-3p组,经尾静脉分别注射AAV-NC和AAV-miR-144-3p后,给予含0.4%β-氨基丙腈的饮用水喂养4周,构建颈动脉夹层大鼠模型。苏木精-伊红染色和EVG染色观察两组颈动脉血管壁的组织结构,免疫荧光染色观察颈动脉血管壁α-平滑肌肌动蛋白(α-SMA)表达。(2)利用慢病毒对A7R5血管平滑肌细胞(A7R5细胞)进行转染并分组:miR-NC组、miR-144-3p mimics组和miR-144-3p抑制剂组。Westernblot法检测各组A7R5细胞中α-SMA、MYH-11、CNN-1、MMP-2、MCP-1、P38、JNK和ERK蛋白及其磷酸化水平;细胞收缩实验检测各组A7R5细胞的收缩能力。结果 miR-144-3p过表达后,颈动脉夹层模型大鼠的颈动脉血管壁弹力纤维排列紊乱,多处发生断裂,血管壁α-SMA水平下降(P<0.05);miR-144-3p过表达后,A7R5细胞的α-SMA、MYH-11和CNN-1蛋白表达均下调(均P<0.05),MMP-2和MCP-1蛋白表达均上调(均P<0.05),细胞收缩功能减弱(P<0.05),细胞形态由排列规则的纺锤形向不规则形转变;miR-144-3p过表达后,A7R5细胞中P38、JNK和ERK蛋白磷酸化水平显著增加,抑制P38可降低MMP-2和MCP-1表达(均P<0.05)。结论miR-144-3p可能参与颈动脉夹层的发生;miR-144-3p通过调控P38 MAPK和JNK信号通路调节血管平滑肌细胞的表型转换。Aim To investigate whether miR-144-3p is involved in carotid artery dissection and the mechanism of regulating the contractive phenotype of vascular smooth muscle cells through mitogenactivated protein kinase(MAPK)signaling pathway.Methods 3-week-old SD male rats were randomly divided into two groups:a AAV-NC group and a AAV-miR-144-3p group.After injection of AAVNC and AAV-miR-144-3p respectively,the rat model of carotid artery dissection was established by feeding 0.4%β-aminopulonitrile in drinking water for 4 weeks.The tissue structure of carotid wall was observed by HE and EVG staining.The expression of aα-smooth muscle actin(a-SMA)was detected by immunofluorescence staining.A7R5 vascular smooth muscle cells were transfected with lentivirus and divided into three groups:a miR-NC group,a miR-144-3p mimics group and a miR-144-3p inhibitor group.The protein expression,includingα-SMA,myosin heavy chain 11(MYH-11),calponin 1(CNN-1),matrix metalloproteinase 2(MMP-2),monocyte chemoattractant protein-1(MCP-1),MAPK signaling pathway molecules(P38,JNK and ERK)and corresponding phosphorylated proteins,was detected by Western blot.Cell contraction assay was performed to measure the contractile ability of vascular smooth muscle cells.Results After miR-144-3p overexpression,the elastic fibers of carotid wall were disordered and broken,and the expression of aα-SMA of carotid artery was decreased significantly(P<0.05).Compared with miR-NC group,the expressions of MYH-11,α-SMA and CNN-1 in A7R5 were downregulated,while the expressions of MMP-2 and MCP-1 were up-regulated in miR-144-3p mimics group(P<0.05).The cell contractile ability was obviously impaired(P<0.05)and the cell morphology changed from regular spindle to irregular shape in miR-144-3p mimics group.In addition,the phosphorylated expression of P38,JNK and ERK in A7R5 was increased significantly in miR-144-3p mimics group,while the inhibition of P38 could reduce the expressions of MMP-2 and MCP-1(P<0.05).Conclusion miR-144-3p may be involved in the occurrence o

关 键 词：miR-144-3p 颈动脉夹层 MAPK信号通路 血管平滑肌细胞 

分 类 号：R743[医药卫生—神经病学与精神病学]