CXCL5通过诱导血管钙化参与颈动脉斑块的形成  

CXCL5 participates in carotid plaque formation by inducing vascular calcification

作　　者：亓明[1] 王磊[1] 张振[1] Qi Ming;Wang Lei;Zhang Zhen(Department of Vascular Surgery,The First Affiliated Hospital of Dalian Medical University,Dalian 116021,Liaoning Province,China)

机构地区：[1]大连医科大学附属第一医院血管外科,辽宁省大连市116021

出　　处：《中国组织工程研究》2024年第2期186-192,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家重点实验室开放基金项目(GZ21104)。

摘　　要：背景:CXC基序趋化因子5(CXC-motif chemokine 5,CXCL5)为上皮细胞衍生的中性粒细胞激活肽,研究发现其可能参与动脉病变。然而,CXCL5在血管钙化中的作用未见报道。目的:探讨CXCL5在颈动脉粥样硬化的血管钙化中的作用。方法:①细胞实验:将小鼠血管平滑肌细胞分成以下各组:成骨培养基组,Vector组(空白质粒转染到细胞中),CXCL5组(CXCL5质粒转染到细胞中),si-NC组(CXCL5阴性对照siRNA转染到细胞中),si-CXCL5组(CXCL5 siRNA转染到细胞中),Vector+LY2157299组和CXCL5+LY2157299组(细胞转染24 h后,将转化生长因子β受体1激酶抑制剂LY2157299加入细胞中)。进行茜素红染色、碱性磷酸酶染色和钙含量测定以评估血管平滑肌细胞成骨分化水平。②动物实验:48只ApoE-/-小鼠随机分成4组:Con+si-NC组、Con+si-CXCL5组、CAS+si-NC组和CAS+si-CXCL5组,前2组不造模,尾静脉注射si-NC或si-CXCL5慢病毒;后2组制备颈动脉粥样硬化模型,尾静脉注射si-NC或si-CXCL5慢病毒。采用Von Kossa染色和免疫组织化学染色评估小鼠颈动脉血管钙化以及CXCL5、转化生长因子β受体1表达情况。结果与结论:①CXCL5组细胞Runt相关转录因子2蛋白水平上调、α-平滑肌肌动蛋白水平下调,si-CXCL5组中的发现与其相反;CXCL5过表达上调了转化生长因子β受体1水平,而CXCL5敲低抑制了转化生长因子β受体1水平。②与Vector组相比,CXCL5组细胞茜素红染色的强度、碱性磷酸酶活性和钙含量显著增加(P<0.05);与si-NC组相比,si-CXCL5组上述2项指标显著降低(P<0.05);当用LY2157299抑制转化生长因子β受体1表达时,CXCL5对平滑肌细胞的成骨转化作用减弱。③与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中CXCL5蛋白表达和血管钙化面积显著增加(P<0.05);与CAS+si-NC组相比,CAS+si-CXCL5组颈动脉中上述2项指标显著降低(P<0.05)。④与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中Runt相关转录因子2蛋白表达显著增加BACKGROUND:CXC motif chemokine 5(CXCL5)is a neutrophil activating peptide derived from epithelial cells,which may be involved in arterial diseases.However,there is yet no report on the effect of CXCL5 in vascular calcification.OBJECTIVE:To explore the role of CXCL5 in the vascular calcification of carotid atherosclerosis(CAS).METHODS:(1)Cytological experiment:Mouse vascular smooth muscle cells(VSMCs)were divided into five groups:osteogenic medium group,Vector group(vector,blank plasmid transfected into VSMCs),CXCL5 group(CXCL5 plasmid transfected into VSMCs),si-NC group(CXCL5 negative control siRNA transfected into VSMCs),si-CXCL5 group(CXCL5 siRNA transfected into VSMCs),Vector+LY2157299 group and CXCL5+LY2157299 group(LY2157299 transferred into the cells 24 hours after cell transfection).Alizarin red staining,alkaline phosphatase staining,and calcium content determination were performed to evaluate the osteogenic differentiation level of VSMCs.(2)Animal experiment:Forty-eight ApoE-/-mice were randomly divided into four groups(n=12 per group):Con+si-NC group,Con+si-CXCL5 group,CAS+si-NC group and CAS+si-CXCL5 group.Animal models were not prepared in the first two groups,in which si-NC or si-CXCL5 lentivirus was injected into the tail vein;carotid atherosclerosis models were made in the latter two groups,in which si-NC or si-CXCL5 lentivirus was injected into the tail vein.Von Kossa staining and immunohistochemical staining were used to evaluate carotid vascular calcification and the expression of CXCL5 and transforming growth factor-βreceptor 1(TGFBR1)in mice.RESULTS AND CONCLUSION:In the CXCL5 group,the protein level of runt-related transcription factor 2(RUNX2)was up-regulated and the level ofα-smooth muscle actin was down-regulated,in contrary to the findings in the si-CXCL5 group.In addition,CXCL5 overexpression upregulated the level of TGFBR1,while CXCL5 knockdown inhibited the level of TGFBR1.Compared with the Vector group,the intensity of alizarin red staining,alkaline phosphatase activity and calcium c

关 键 词：CXC基序趋化因子5 颈动脉粥样硬化 血管钙化 血管平滑肌细胞 转化生长因子β受体1 

分 类 号：R446[医药卫生—诊断学] R318[医药卫生—临床医学] R496