摘要
目的探讨内皮祖细胞条件培养基抑制血管紧张素Ⅱ诱导的血管平滑肌细胞增殖的分子机制。方法制备早期内皮祖细胞条件培养基,经降钙素基因相关肽抗体预处理内皮祖细胞条件培养基或特异性阻断剂阻断内皮祖细胞降钙素基因相关肽受体后,采用免疫印迹、逆转录聚合酶链反应、间接免疫荧光观察内皮祖细胞条件培养基对血管紧张素Ⅱ所致血管平滑肌细胞MAPK、NF.KB信号转导通路和原癌基因c-myc、c-los表达的影响。结果降钙素基因相关肽阻断后,早期内皮祖细胞条件培养基对血管紧张素Ⅱ诱导的血管平滑肌细胞MAPK(P38、JNK)、核因子-κB(亚单位P65)信号转导通路的活化以及原癌基因c-myc、c-fos表达的抑制能力较对照组明显降低。结论内皮祖细胞通过旁分泌降钙素基因相关肽抑制血管紧张素Ⅱ诱导的血管平滑肌细胞增殖,其机制可能与其抑制血管平滑肌细胞MAJPK(P38、JNK)、核因子-κB信号转导通路的活化以及原癌基因c-myc和c-fos表达有关。
Objective To investigate the molecular mechanism of endothelial progenitor cells (EPCs) conditioned medium to inhibit angiotensin Ⅱ -induced proliferation of vascular smooth muscle ceils (VSMCs). Methods First,early endothelial progenitor cell conditional medium (E-EPC-CM)was pre-incubated with functional blocking antibodies against calcitonin gene related peptide (CGRP)or VSMCs was pretreated with CGRP837 (CGRP receptor antagonist)before VSMCs were pretreated with conditional medium;then Western blot,reverse transcriptase polymerase chain reaction and indirect immunofluorescenee were used to observe the activation of mitogen-activated protein kinase (MAPK)and nuclear transcription factor-κB (NF-κappaB)signal transduction pathways and the effect of proto-oncogene c-myc and e-los in angiotensin Ⅱ -induced VSMCs. Results After pretreatrnent with anti-CGRP antibody and CGRP837,the inhibitory effect of E-EPC-CM on angiotensin Ⅱ-induced activation of of p38,JNK of MAPKs and NF -κappaB subunit p65 (or nuclear translocation of p65)signal transduction pathway and the expression of protooncogene c-myc and c-los was significantly lower than that of the control group.Conclusion EPCs inhibit the proliferation of VSMCs induced by angiotensin Ⅱ by paracrine CGRP,the mechanism may be related to the inhibition of MAPK and NF-κappaB subunit p65 and expression of proto-oncogenes C-myc and C-fos.
作者
方立
黄钦
黄芳
李迎
张银妆
FANG Li;HUANG Qin;HUANG Fang;LI ]qng;ZHANG Yin-zhuang(Department of Cardiology,The First Hospital of Changsha,Changsha 410008,China)
出处
《中国心血管病研究》
CAS
2019年第1期82-86,共5页
Chinese Journal of Cardiovascular Research
基金
湖南省自然科学基金(2018JJ6127)
湖南省卫生计生委科研计划B类课题项目(B20180471)
长沙市科技局项目(K15ZD022-33、Kq1701007).
关键词
内皮祖细胞
血管平滑肌细胞
血管紧张素Ⅱ
增殖
降钙素基因相关肽
Endothelial progenitor cell (EPCs)
Vascular smooth muscle ceils (VSMCs)
Ang Ⅱ
Proliferation
Calcitonin gene-related peptide
