摘要
目的:探究β-淀粉样蛋白1-40(Aβ1-40)对血管平滑肌细胞(VSMCs)的炎症反应、活力、迁移能力及表型转化的影响,并分析其机制。方法:以不同浓度的Aβ1-40干预VSMCs适当时间。用CCK-8和Transwell实验评估细胞的活力及迁移能力;用Western blot检测细胞中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)等炎症因子水平,VSMCs表型转化标志蛋白α-平滑肌肌动蛋白(α-SMA)、骨桥蛋白(OPN)和Krüppel样因子4(KLF4)的表达,以及丝裂原活化蛋白激酶(MAPKs)信号通路相关蛋白p-p38 MAPK、p-ERK1/2和p-JNK的蛋白水平。结果:在Aβ1-40的作用下,VSMCs中炎症因子IL-1β和TNF-α水平显著升高,表型转化相关蛋白表达发生改变,α-SMA表达水平显著下降,而OPN和KLF4的表达水平显著升高(P<0.05);且在一定浓度范围内,Aβ1-40可提高VSMCs的活力和迁移能力;另外,Aβ1-40处理可显著增加VSMCs中p-p38 MAPK、p-ERK1/2和p-JNK的蛋白水平(P<0.05)。应用MAPKs相应的抑制剂可显著降低IL-1β和TNF-α表达水平(P<0.05),并抑制细胞表型转化,即α-SMA表达水平显著升高,OPN和KLF4表达水平显著下降(P<0.05)。结论:Aβ1-40可通过激活MAPKs通路诱导VSMCs发生炎症反应和表型转化。
AIM:To investigate the effects of amyolidβ-protein 1-40(Aβ1-40)on inflammation,viability,migration and phenotypic switching in vascular smooth muscle cells(VSMCs),and to analyze the underlying mecha⁃nisms.METHODS:The VSMCs were treated with Aβ1-40 at different concentration gradients for appropriate time.CCK-8 and Transwell assays were performed to evaluate the viability and migration ability of VSMCs.The levels of inflam⁃matory factors including interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α),phenotypic switching-related pro⁃teins includingα⁃smooth muscle actin(α⁃SMA),osteopontin(OPN)and Krüppel⁃like factor 4(KLF4),and mitogen-ac⁃tivated protein kinases(MAPKs)signaling pathway-related proteins including p-p38 MAPK,p-ERK1/2 and p-JNK were determined by Western bolt.RESULTS:After Aβ1-40 treatment,the levels of inflammatory factors IL-1βand TNF-αin the VSMCs were significantly increased(P<0.05),and the expression of phenotypic switching-related proteins was al⁃tered,as indicated by down-regulation ofα⁃SMA and up-regulation of OPN and KLF4(P<0.05).Treatment with Aβ1-40 within a certain concentration range promoted the viability and migration of the VSMCs.In addition,the protein levels of p-p38 MAPK,p-ERK1/2 and p-JNK were significantly increased by Aβ1-40 treatment(P<0.05).Furthermore,pretreat⁃ment with specific inhibitors of MAPKs pathway significantly reduced the levels of IL-1βand TNF-α(P<0.05),and in⁃hibited the phenotypic switching,as indicated by up-regulation ofα⁃SMA and down-regulation of OPN and KLF4(P<0.05).CONCLUSION:Treatment with Aβ1-40 induces the inflammation and phenotypic switching in VSMCs via acti⁃vation of MAPKs signaling pathway.
作者
张杰
黄兴晓
池菊芳
ZHANG Jie;HUANG Xing-xiao;CHI Ju-fang(Department of Cardiology,Shaoxing People's Hospital(Shaoxing Hospital,Zhejiang University School of Medicine),Shaoxing 312000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第11期1966-1971,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81873120)
浙江省医药卫生科研资助项目(No.2018KY172)。
