生理性周期性张应变通过激活AMPK磷酸化抑制血管平滑肌细胞迁移

目的 探讨细胞能量代谢的关键调节因子 AMP激活的蛋白激酶AMPK在血管平滑肌细胞（vascular smooth muscle cells, VSMCs）响应生理性周期性张应变力学刺激后对VSMCs迁移的影响。方法 采用 Flexcell-5000T体外细胞张应变加载系统,对大鼠原代培养的 VSMCs 施加10%幅度、1.25 Hz 频率的周期性张应变,模拟VSMCs在体内的生理性力学环境;以未加载周期性张应变的静态细胞为对照组,Western blotting 检测 VSMCs的 p-AMPK蛋白表达;划痕实验检测 VSMCs 迁移功能。结果 与静态组的细胞相比,生理性周期性张应变加载24 h后显著减少划痕愈合面积,提示生理性周期性张应变抑制VSMCs迁移;生理性周期性张应变加载3 h后,VSMCs的p-AMPK蛋白表达显著升高,而加载24 h后p-AMPK蛋白表达显著降低。在生理性周期性张应变加载条件下,孵育AMPK抑制剂可以在张应变加载3 h后显著降低 p-AMPK蛋白表达,而在张应变加载24 h后显著促进VSMCs迁移;在静态条件下孵育AMPK激活剂 AICAR 3 h后显著诱导p-AMPK蛋白表达,孵育24 h后显著抑制VSMCs迁移;提示p-AMPK蛋白表达参与调控VSMCs迁移。结论 生理性周期性张应变能通过激活p-AMPK蛋白表达,进而抑制VSMCs迁移,提示生理性周期性张应变调控VSMCs迁移对维持血管稳态具有重要意义。     

FOXO1参与高血压条件下异常张应变诱导的血管平滑肌细胞增殖

目的 探讨高血压条件下异常升高的周期性张应变刺激对血管平滑肌细胞（vascular smooth muscle cells, VSMCs）增殖的影响,以及Forkhead转录因子1（FOXO1）在其中可能的作用。方法 构建腹主动脉缩窄高血压大鼠模型,并以假手术组为对照,应用FX-4000T体外周期性张应变加载系统,分别对VSMCs施加5%的生理性张应变和15%的高血压病理性张应变。Western blot检测VSMCs的FOXO1及p-FOXO1表达水平,BrdU法检测VSMCs 增殖活性。RNA干扰技术抑制VSMCs的FOXO1表达,检测FOXO1、p-FOXO1表达以及VSMCs增殖活性变化。结果 腹主动脉缩窄术后 2和4周,大鼠血压较假手术大鼠明显增高。与假手术大鼠相比,高血压大鼠血管壁细胞增殖活性明显增高,同时 FOXO1及 p-FOXO1表达水平也显著性升高。细胞实验表明,与5%张应变组相比,15%张应变加载显著上调VSMCs的FOXO1、p-FOXO1表达水平,以及VSMCs增殖活性。静态条件下RNA干扰抑制VSMCs的FOXO1及p-FOXO1表达,VSMCs的增殖活性明显降低。结论 高血压病理条件下,异常增高的周期性张应变可能通过促进 FOXO1表达和磷酸化诱导VSMCs增殖。以动物模型观察现象,在细胞分子水平探讨机制,旨在明确FOXO1在高血压血管重建中的作用及其力学生物学机制,为阐明高血压血管重建的发病机理和药物治疗靶标的研究提供新的实验依据。     

机械张应变诱导的大鼠血管平滑肌细胞磷脂酰肌醇3激酶/蛋白激酶B信号通路相关基因的筛选

目的 筛选机械张应变诱导的大鼠血管平滑肌细胞(VSMCs)磷脂酰肌醇3激酶/蛋白激酶B(PI3K/Akt)信号通路的相关基因.方法 应用FX-4000T 细胞应变加载系统,对大鼠主动脉VSMCs施加1 Hz、10 % 张应变,Western blotting检测细胞在不同加载时间下Akt磷酸化水平的变化;用抑制消减杂交法(SSH) ,筛选给予PI3K的抑制剂Wortmannin和给予DMSO 两组细胞在加载张应变24 h后的差异表达基因片段,得到的消减杂交产物连接到T 载体上,经过蓝白斑筛选,对所有阳性克隆进行菌液PCR筛选及测序,应用BLASTN进行序列比对.结果 机械张应变改变了VSMCs磷酸化Akt水平;SSH所获得的54个克隆随机挑选30个送测序,得到10个不同差异表达序列标签(EST),发现6个EST代表的大鼠基因可能与机械张应变诱导的VSMCs PI3K/Akt信号通路相关,它们是:High mobility group box 1(HMGB1), Neural precursor cell expressed(Nedd4a ), Cyclin-dependent kinase 5(CDK5), Histone deacetylase 3(HDAC3), Ubiquitin-like 1(Uble1a), Heterogeneous nuclear ribonucleoprotein H1(hnRNP).结论 SSH有效筛选到了机械张应变诱导的VSMCs PI3K/Akt信号通路的相关基因,为进一步研究机械张应变诱导的血管病理生理改变提供了资料.     

长链非编码RNA XR007793在病理性高张应变诱导平滑肌细胞增殖中的作用

目的探讨高血压条件下异常升高的周期性张应变刺激对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖活性的影响以及VSMCs中一种长链非编码RNA(long non-coding RNA,lncRNA)XR007793在其中可能的作用。方法应用体外周期性张应变加载系统Flexcell-4000对VSMCs分别施加5%生理性张应变和15%病理性高张应变,加载频率为1.25 Hz,加载时间24 h。荧光实时定量PCR检测XR007793及其共表达基因:信号转导与转录激活因子2(STAT2)、细胞分裂相关蛋白8(CDCA8)、原癌基因LMO2和干扰素调节因子(IRF7)的表达变化,Western bloting方法检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)表达变化,RNA干扰技术抑制VSMCs的XR007793表达,静态条件下流式细胞术检测VSMCs周期变化,牵拉条件下Brdu检测VSMCs增值变化情况。结果与5%生理性张应变组相比,15%病理性高张应变显著下调XR007793表达水平,促进VSMCs增值活性并上调STAT2和CDCA8的表达水平。静态条件下干扰XR-007793,VSMC增殖水平显著上升。高周期性张应变条件下干扰XR-007793,VSMC增殖水平上升,CDCA8表达水平上升。结论病理性高张应变可能通过降低XR007793表达来影响CDCA8表达变化,进而调控VSMCs增殖功能变化过程。研究结果为阐明高血压条件下血管重建机制和药物治疗靶标的研究提供新的实验依据。     

三七皂苷Rg1抑制脂多糖诱导小胶质细胞激活的机制研究

目的:探讨中药有效成分三七皂苷Rg1(Ginsenoside Rg1,Rg1)对抑制脂多糖(lipopolysaccharide,LPS)诱导的小胶质细胞株BV-2细胞激活的机制。方法:用LPS刺激BV-2细胞构建激活模型,采用四甲基偶氮唑蓝比色法(MTT)检测Rg1对BV-2细胞的活力影响,蛋白质免疫印迹(Western Blot)方法检测不同浓度Rg1(10、20和40μmol/L)对磷酸化的核因子-κB抑制蛋白-α(inhibitorκB-α,IκB-α)和反应结合蛋白(cAMP-responseelement binding protein,CREB)以及促分裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)家族的细胞外信号调节激酶(extracellular signal-regulated kinase 1/2,ERK1/2)、c-Jun氨基端激酶(c-Jun N-terminal kinase,JNK)和p38促分裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)等细胞信号通路蛋白的表达及其变化规律。结果:不同浓度的Rg1明显抑制了LPS诱导的磷酸化IκB-α和CREB蛋白表达以及MAPKs通路(ERK1/2,JNK,p38 MAPK)磷酸化蛋白表达,并且对p38 MAPK表达的影响呈剂量依赖性。结论:Rg1可能通过抑制MAPKs的磷酸化来调控LPS诱导的小胶质细胞株BV-2细胞激活,发挥其神经抗炎的作用。     

周期性张应变调控血管平滑肌细胞黏附血小板微体及其在自噬中的作用

目的探讨周期性张应变力学刺激对血管平滑肌细胞(vascular smooth muscle cells,VSMCs)与血小板微体(platelet-derived microparticles,PMPs)黏附能力的影响,以及黏附的PMPs对VSMCs自噬的调控作用。方法应用FX-5000T张应变加载系统,对体外培养VSMCs施加5%幅度的生理性张应变和15%幅度的高张应变;应用流式细胞术检测不同张应变作用的VSMCs与PMPs的黏附;免疫荧光检测PMPs刺激24 h后自噬标志分子微管相关蛋白轻链3(autophagy microtubule associated protein light chain 3,LC3)的表达水平; Western blotting检测PMPs刺激24 h后VSMCs自噬相关蛋白(autophagy related protein,Atg)的表达水平。结果与5%生理性张应变加载相比,15%高张应变加载24 h能显著增强VSMCs与PMPs的黏附水平,提示高张应变促进PMPs与VSMCs的黏附。免疫荧光和Western blotting结果显示,PMPs刺激可显著上升VSMCs中自噬标志蛋白LC3表达,同时Western blotting检测到PMPs刺激后Atg5、Atg7、Atg12蛋白表达水平显著上升。结论高张应变可以促进VSMCs黏附PMPs,黏附的PMPs可能通过增加Atg5、Atg7、Atg12、LC3表达,从而增强VSMCs自噬。     

FKN对人外周血单个核细胞PKC-ERKl/2活化和TNF-α表达的影响

目的 探索FKN-CX3CR1在单个核细胞中可能存在的信号传导途径及促进动脉粥样硬化形成的机制,并探讨蛋白激酶C（PKC）在其中作用。方法 （1）用Ficoll密度梯度离心法分离抗凝人外周血单个核细胞。（2）将每份提取的单个核细胞分为4组：对照组、FKN组、Ro31-8220（PKC特异性阻断剂）和PD98059（ERK1/2特异性阻断剂）组。（3）用Western blot法检测单个核细胞中磷酸化ERK1/2表达。（4）用ELISA法检测培养液中TNF-α的表达。结果 （1）FKN组磷酸化的ERK1/2和TNF－α表达较对照组显著增多（P<0.05）。（2）Ro31-8220组磷酸化的ERK1/2和TNF－α表达较FKN组显著减少（P<0.05）。结论 FKN－CX3CR1可能通过PKC/ERK途径诱导单个核细胞TNF-α的表达,最终促进动脉粥样硬化的形成和进展。     

生理性张应变通过增加核骨架蛋白Emerin表达抑制血管平滑肌细胞凋亡

目的探讨细胞核骨架蛋白Emerin及其调控的转录因子在感受张应变力学刺激影响血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡中的作用。方法应用FX-5000T张应变加载系统,对体外培养的VSMCs施加5%幅度、1.25 Hz频率生理性张应变,以静止组为对照;应用cleaved-caspase3 ELISA试剂盒检测VSMCs凋亡水平,Western blotting检测VSMCs细胞核骨架蛋白Emerin蛋白表达水平。静态条件下,RNA干扰抑制VSMCs的Emerin表达,Protein/DNA芯片检测345种转录因子活性;将特异性干扰Emerin后活性发生明显变化(上调或下调超过2倍)的转录因子进行IPA(Ingenurity Pathway Analysis)信息学分析,筛选与凋亡功能相关的转录因子;染色质免疫共沉淀(chromatin immunoprecipitation,CHIP)结合q PCR检测特异性干扰Emerin对其与两种转录因子motif区域结合能力的影响。结果与静止组相比,5%生理性张应变加载24 h后,VSMCs凋亡水平显著降低,提示生理性张应变对细胞具有保护作用;5%张应变作用6、12和24 h均显著增加VSMCs的Emerin表达水平。静态条件下RNA干扰抑制Emerin表达,VSMCs凋亡水平显著增加,且10种参与细胞凋亡功能调控的转录因子活性显著上调(2倍以上),包括CREB-BP1、p300、p55、MAX、NRF-1、STAT1、STAT3、TEF1、TR和BZP;CHIP-q PCR结果显示,Emerin特异性干扰可以显著降低Emerin与STAT家族的两个成员STAT1和STAT3的motif区域结合能力。结论生理性张应变可能通过增加核骨架蛋白Emerin表达而调控Emerin与STAT1、STAT3等多种凋亡相关转录因子motif区域结合,进而调控转录因子活性影响VSMCs凋亡。探讨张应变力学刺激调控VSMCs功能的力学生物学分子机制,对揭示血管生理稳态维持和血管病理重建的分子机制具有一定意义。     

精氨酸加压素对大鼠视前区GABA_A受体亚单位磷酸化的影响

目的:研究精氨酸加压素(AVP)对大鼠视前区γ-氨基丁酸(GABA)A型受体(GABA_A受体)亚单位(α、β和γ2)表达和磷酸化的影响。方法:实验分为对照组、AVP组、V1a受体抑制剂+AVP组和V1a受体抑制剂组(均n=10);腹腔注射AVP或V1a受体抑制剂0.5 h后,采用RT-qPCR和Western blot法检测视前区GABA_A受体亚单位(α、β和γ2)表达及磷酸化的变化。结果:与对照组相比,AVP或V1a受体抑制剂组大鼠视前区GABA_A受体亚单位表达均无显著变化;AVP能显著上调视前区GABA_A受体γ2亚单位的磷酸化水平(P0.05);AVP显著增加蛋白激酶C(PKC)和钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)表达和磷酸化(P0.01)。结论:外源性AVP不影响GABA_A受体亚单位(α、β和γ2)表达,但主要通过V1a受体激活PKC和CaMKⅡ,影响γ2亚单位磷酸化水平,从而调制视前区GABA_A受体介导的抑制性突触传递。     

Mad1 kinetochore recruitment by Mps1-mediated phosphorylation of Bub1 signals the spindle checkpoint

The spindle checkpoint is a conserved signaling pathway that ensures genomic integrity by preventing cell division when chromosomes are not correctly attached to the spindle. Checkpoint activation depends on the hierarchical recruitment of checkpoint proteins to generate a catalytic platform at the kinetochore. Although Mad1 kinetochore localization is the key regulatory downstream event in this cascade, its receptor and mechanism of recruitment have not been conclusively identified. Here, we demonstrate that Mad1 kinetochore association in budding yeast is mediated by phosphorylation of a region within the Bub1 checkpoint protein by the conserved protein kinase Mps1. Tethering this region of Bub1 to kinetochores bypasses the checkpoint requirement for Mps1-mediated kinetochore recruitment of upstream checkpoint proteins. The Mad1 interaction with Bub1 and kinetochores can be reconstituted in the presence of Mps1 and Mad2. Together, this work reveals a critical mechanism that determines kinetochore activation of the spindle checkpoint.     

DDX3和酪蛋白激酶1ε在肌萎缩侧索硬化症转基因鼠脑干中的表达变化

目的 检测DDX3和酪蛋白激酶1ε(CK1ε)在肌萎缩侧索硬化症(ALS)转基因鼠脑干中的表达变化,探讨DDX3和CK1ε在ALS脑干运动神经元变性中的作用。 方法 选取ALS转基因鼠33只,分别于发病早期（95d）、中期（108d）和晚期（122d）3个时间点剥离脑干,应用RT-PCR、Western blotting和免疫荧光染色技术分别检测ALS转基因鼠脑干组织中DDX3和CK1ε的表达规律,在脑干运动核团舌下神经（12N）和面神经（7N）中阳性细胞的分布特点及细胞定位,每组均选择相同数量的同窝野生型鼠作为对照。 结果 RT-PCR和Western blotting结果显示,与同窝野生型鼠相比,ALS转基因鼠脑干组织中DDX3和CK1εmRNA于95 d、108d、122d表达均无明显变化,DDX3和CK1ε蛋白在95d和108d表达上调,122d表达下调(P<0.01, P<0.001)。免疫荧光结果显示,在ALS鼠和野生型鼠脑干的12N和7N区域均可检测到DDX3和CK1ε阳性细胞,DDX3和CK1ε表达在神经元,在星形胶质细胞不表达。ALS鼠和野生型鼠DDX3和CK1ε免疫反应性具有差异。 结论 DDX3和CK1ε在脑干中的表达异常与ALS发病密切相关。     

The initial phase of chromosome condensation requires Cdk1-mediated phosphorylation of the CAP-D3 subunit of condensin II

The cell cycle transition from interphase into mitosis is best characterized by the appearance of condensed chromosomes that become microscopically visible as thread-like structures in nuclei. Biochemically, launching the mitotic program requires the activation of the mitotic cyclin-dependent kinase Cdk1 (cyclin-dependent kinase 1), but whether and how Cdk1 triggers chromosome assembly at mitotic entry are not well understood. Here we report that mitotic chromosome assembly in prophase depends on Cdk1-mediated phosphorylation of the condensin II complex. We identified Thr 1415 of the CAP-D3 subunit as a Cdk1 phosphorylation site, which proved crucial as it was required for the Polo kinase Plk1 (Polo-like kinase 1) to localize to chromosome axes through binding to CAP-D3 and thereby hyperphosphorylate the condensin II complex. Live-cell imaging analysis of cells carrying nonphosphorylatable CAP-D3 mutants in place of endogenous protein suggested that phosphorylation of Thr 1415 is required for timely chromosome condensation during prophase, and that the Plk1-mediated phosphorylation of condensin II facilitates its ability to assemble chromosomes properly. These observations provide an explanation for how Cdk1 induces chromosome assembly in cells entering mitosis, and underscore the significance of the cooperative action of Plk1 with Cdk1.     

SASH1通过MAP2K2和MAP4K4与ERK信号通路交互作用

目的 探讨新候选抑癌基因SASH1与细胞外调节蛋白激酶(ERK)信号通路的两个关键分子MAP2K2和MAP4K4的蛋白-蛋白相互作用关系.方法 用Xho Ⅰ和HpaⅠ构建SBP-Flag-SASH1-pBABE-puro反转录病毒载体,转染HEK-293T细胞,通过嘌呤霉素筛选出稳定表达外源性SASH1基因的细胞系.Western blot检测外源性Flag-SASH1蛋白表达,利用pull-down实验、质谱技术、免疫沉淀鉴定和分析与SASH1结合的并可能调节细胞增殖、转移和凋亡等的关键蛋白.用SASH1-siRNA1和SASH1-siRNA2分别转染MDA-MB-231细胞系,以空白组和Negative-siRNA组为对照.72 h后Western blot检测SASH1的干扰效果和P-ERK1/2水平.结果 成功构建稳定表达SBP-Flag-SASH1-pBABE-puro重组质粒的HEK-293T细胞系,SASH1与MAP2K2和MAP4K4结合并发生相互作用.SASH1-siRNA有效抑制MDA-MB-231细胞SASH1蛋白表达(P＜0.05),且P-ERK1/2在SASH1抑制组表达增加.结论 MAP2K2和MAP4K4是SASH1的重要的候选结合蛋白,SASH1可能通过与其直接或间接结合串话ERK信号传导通路,进而调节细胞增殖和迁移等细胞生物学功能.     

Snf1 kinase complexes with different beta subunits display stress-dependent preferences for the three Snf1-activating kinases

Three upstream kinases, Pak1, Tos3 and Elm1, are able to activate the Snf1 kinase. Since the Snf1 kinase itself assembles into three complexes that differ in their beta subunit identity, the possibility exists that each upstream kinase might be dedicated to a single isoform of the Snf1 kinase. To test this dedicated activator hypothesis, we generated a series of yeast strains that lacked different combinations of upstream kinases and beta subunits. Cells expressing only one of the three upstream kinases exhibited distinct abilities to activate Snf1, depending on the beta subunit present in the Snf1 kinase complex and the stress imposed on the cells. Pak1 and Gal83 were the most promiscuous. Pak1 was able to activate all three isoforms of the Snf1 kinase under all stress conditions tested. The Gal83 isoform of Snf1 was able to be activated by any of the three upstream kinases under aerobic growth conditions but showed a preference for Pak1 during growth on raffinose. Our results indicate that the three Snf1-activating kinases are not dedicated to specific isoforms of the Snf1 kinase. Instead, the different isoforms of the Snf1 kinase display stress-dependent preferences for the Pak1, Tos3 and Elm1 kinases.     

The toxicity of dysregulated Plk1 activity revealed by its suppressor mutations

Polo-like kinase 1 (Plk1) is a mitotic kinase that has multiple functions throughout the cell cycle. Catalytic activation of Plk1 is known to be regulated by phosphorylation of the kinase domain, including Thr210, and by releasing the kinase domain from its inhibitory polo-box domain. However, how Plk1 is activated to fulfill its proper roles, in time and space, is not well understood. In this study, we unintentionally found that the expression of a constitutively active form of human Plk1 is toxic to bacterial cells, such that cells contained point mutations that alleviate the kinase activity. Structural prediction revealed that these mutations are adjacent to the amino acids supporting the kinase activity. When human cells express these mutants, we found decreased levels of Plk1's substrate phosphorylation, resulting in mitotic defects. Moreover, unlike in bacterial cells, the expression of activated Plk1 mutants did not affect cell proliferation in human cells unless localized at the right place in mitosis. Our observations identified new suppressor mutations and underscored the importance of spatiotemporal regulation in Plk1, providing a basis for how we might intervene in this kinase for therapeutic purpose in human cells.     

Inflammatory mediators and islet β-cell failure: a link between type 1 and type 2 diabetes

Pancreatic islet beta-cell death occurs in type 1 and 2 diabetes mellitus, leading to absolute or relative insulin deficiency. beta-cell death in type 1 diabetes is due predominantly to autoimmunity. In type 2 diabetes beta-cell death occurs as the combined consequence of increased circulating glucose and saturated fatty acids together with adipocyte secreted factors and chronic activation of the innate immune system. In both diabetes types intra-islet inflammatory mediators seem to trigger a final common pathway leading to beta-cell apoptosis. Therefore anti-inflammatory therapeutic approaches designed to block beta-cell apoptosis could be a significant new development in type 1 and 2 diabetes.     

Curcumin attenuates ethanol-induced toxicity in HT22 hippocampal cells by activating mitogen-activated protein kinase phosphatase-1

Ethanol causes neurotoxicity through formation of reactive oxygen species and activation of mitogen-activated protein kinase (MAPK) pathways. MAPK phosphatase-1 (MKP-1) is one of the phosphatases responsible for dephosphorylation/deactivation of MAPKs. In this report, we examined the potential involvement of MKP-1 in cytoprotective effects of the well-known antioxidant curcumin. In HT22 hippocampal cells, ethanol caused cell death and activation of p38 MAPK and other two kinases. Blockage of p38 MAPK by its inhibitor protected HT22 cells against ethanol-induced toxicity. Curcumin attenuated ethanol-induced cell death, inhibited activation of p38 MAPK, and activated MKP-1. In HT22 cells transiently transfected with small interfering RNA against MKP-1, curcumin failed to inhibit ethanol-induced activation of p38 MAPK and to protect HT22 cells from ethanol-induced toxicity. Our results suggest that curcumin can attenuate ethanol-induced neurotoxicity by activating MKP-1 which acts as the negative regulator of p38 MAPK. This novel pathway may contribute to and explain at least one of the cytoprotective actions of curcumin.     

Plk1调节c-Abl表达机制的初步研究

目的 研究Polo样激酶1(Polo-like kinase1,Plk1)对非受体酪氨酸激酶(cellular-abelsongene,c-Abl)表达水平的调控及其作用机制.方法 在293T细胞中过表达带有不同标签的Plk1,研究随着外源Plk1表达量的增加,内源c-Abl表达是否变化.分别通过放线菌酮翻译抑制实验和荧光定量PCR实验,研究Plk1是通过影响c-Abl蛋白稳定性还是转录活性来调节其表达.结果 细胞内c-Abl的表达随着外源Plk1的表达量的增加而增加,Plk1不影响c-Abl蛋白稳定性,但可调节c-Abl转录水平.结论 Plk1可在转录水平调节c-Abl表达.     

IL-1通过PKC/MAPK通路上调泡沫细胞ACAT-1的表达及活性

 摘要 目的 研究白细胞介素1(Interleukin 1, IL-1)是否通过蛋白激酶C/丝裂原激活蛋白激酶（PKC/MAPK）信号通路上调泡沫细胞中脂酰CoA胆固醇酯酰转移酶-1（ACAT-1）的表达及活性。方法 复苏并培养人单核细胞株THP-1细胞,与200nM乙酸肉豆蔻佛波酯（PMA）共孵育48h,再与氧化低密度脂蛋白（Ox-LDL）共孵育24h,油红O染色观察细胞质内脂质沉积;检测泡沫细胞、泡沫细胞加IL-1及泡沫细胞加IL-1/IL-1单克隆抗体三组细胞中PKC和MAPK的活性;在三组细胞中分别加入PKC和MAPK抑制剂,分别以Western blot及液相闪烁计数法检测ACAT-1的蛋白表达及酶活性。结果 与PMA共孵育48h后,THP-1细胞逐渐伸出突起,由圆形、悬浮式生长转变为多角形、梭形,呈阿米巴样贴壁生长;经Ox-LDL诱导24h后,油红O染色胞质内可见大量吞噬的脂质小滴。与泡沫细胞组相比,泡沫细胞加IL-1组PKC活性（P<0.05）、MAPK活性（P<0.05）、ACAT-1蛋白表达及活性增加（P<0.05）;PKC抑制剂和MAPK抑制剂能显著抑制IL-1上调ACAT-1表达（P<0.05）及活性（P<0.05）的作用。结论 IL-1上调泡沫细胞ACAT-1表达及活性的作用是通过PKC/MAPK信号通路途径实现的。