摘要
目的探讨血红素加氧酶-1(HO-1)在血管平滑肌细胞(VSMCs)泡沫样变过程中的作用及可能的机制。方法用组织贴块法体外培养C57BL/6J背景野生型小鼠及TLR4基因敲除(TLR4^-/-)小鼠主动脉VSMCs;用氧化型低密度脂蛋白(ox-LDL)刺激VSMCs泡沫样变构建细胞模型,利用HO-1激动剂钴原卟啉(CoPPIX)及抑制剂锌卟啉(ZnPPIX)刺激模型细胞;用油红O染色观察VSMCs内脂质聚积情况;酶法检测VSMCs内胆固醇水平;蛋白免疫印迹(Western blot)检测VSMCs中HO-1、Toll样受体4(TLR4)表达情况;酶联免疫吸附试验(ELISA)法检测VSMCs培养液中炎症因子白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的表达情况,以探索HO-1在VSMCs泡沫样变过程中的具体作用。结果ox-LDL刺激VSMCs48h显著诱导VSMCs泡沫样变;ox-LDL诱导VSMCs泡沫样变过程中伴随着TLR4及其下游炎症因子IL-6、TNF-α表达的上调;而对于TLR4^-/-型VSMCs,ox-LDL刺激并不能有效诱导VSMCs泡沫样变及影响IL-6、TNF-α表达;激活HO-1显著抑制ox-LDL诱导的VSMCs泡沫样变,伴随着TLR4及其下游炎症因子IL-6、TNF-α表达的下调;而抑制HO-1进一步促进ox-LDL诱导的VSMCs泡沫样变,同时伴随着TLR4及其下游炎症因子IL-6、TNF-α表达的上调。结论HO-1的激活可通过下调TLR4介导的炎性反应抑制VSMCs泡沫细胞的形成。
Objective To investigate the role and possible mechanism of heme oxygenase-1(HO-1)in the foam cell formation of vascular smooth muscle cell(VSMC).Methods Primary VSMCs isolated from the thoracic aorta of the wild-type mice C57BL/6J and Toll like recepter 4(TLR4)knockout(TLR4^-/-)mice were cultured in vitro by tissue patch method.Oxidized low density lipoprotein(ox-LDL)was used to induce the VSMC foaming.In this process,CoPPIX and ZnPPIX were respectively used to promote or inhibit the HO-1 expression.Red oil O staining was used to observe the intracellular lipid deposition;cholesterol level in VSMCs was detected by enzymatic assay;the HO-1 and TLR4 expression levels were detected by Western blot;the levels of IL-6 and TNF-αwere quantified by using ELISA.Results ox-LDL stimulated VSMCs for 48 h and induced foam-like changes successfully,and this process was accompanied by the up-regulation of the TLR4,IL-6 and TNF-α.However,in TLR4^-/-VSMCs,ox-LDL failed to induce the VSMC foaming changes and affect the expression levels of IL-6,TNF-α.Up-regulation of HO-1 inhibited the VSMC foaming changes which induced by ox-LDL,and down-regulated the expression levels of TLR4,IL-6 and TNF-α,while down-regulation of HO-1 may play the opposite role.Conclusion The up-regulation of HO-1 could inhibit the VSMC foaming changes which induced by ox-LDL via suppressing TLR4-mediated inflammation.
作者
尹延伟
孙倩倩
沈勇
胡爱民
王琦
陈大伟
赵炫柱
石进
YIN Yanwei;SUN Qianqian;SHEN Yong;HU Aimin;WANG Qi;CHEN Dawei;ZHAO Xuanzhu;SHI Jin(Department of Neurology,Air Force Medical Center,Beijing 100142,China;The Ninth Sanatorium of Beijing Garrison Command of Weishu District,Beijing 100021,China;Comprehensive Ward,Hebei Provincial Corps Hospital of PAP,Shijiazhuang,Hebei 050081,China;Department of Emergency,Air Force Medical Center,Beijing 100142,China)
出处
《重庆医学》
CAS
2019年第16期2701-2704,共4页
Chongqing medicine
基金
国家自然科学基金项目(81500356)
北京市科技新星计划基金项目(Z171100001117094)
