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THP-1巨噬细胞热休克处理对单核细胞超化蛋白-1和白细胞介素表达的影响 被引量:6

The effect of heat shock on the expression of MCP-1 and IL-8 in THP-1 cells
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摘要 目的:探讨热休克处理对THP-1巨噬细胞表达单核细胞趋化蛋白-1(MCP-1)、白细胞介素-8(IL-8)的影响以及热休克蛋白70(HSP70)、TLR4/P38MAPK和TLR4/NF-κB在其中的作用。方法:实验前用佛波酯孵育THP-1细胞,使其诱导分化成巨噬细胞,换无血清培养基培养后加处理因素。实验分成A组:常温对照组;B组:热休克处理组。THP-1巨噬细胞在42℃水浴受热1h,37℃恢复6h;C组:热休克+抗TLR4抗体组,THP-1巨噬细胞加入anti-TLR42h后同B组;D组:热休克+P38MAPK抑制剂组,THP-1巨噬细胞加入P38MAPK特异性抑制剂SB20358030min后同B组;E组:热休克+NF-κB抑制剂组:THP-1巨噬细胞加入NF-κB特异性抑制剂PDTC30min后同B组;F组:热休克+抗HSP702h抗体组,THP-1巨噬细胞加入anti-HSP702h后同B组。用RT-PCR或westernblot或ELISA检测各组细胞HSP70、TLR4、MCP-1、IL-8mRNA或蛋白的表达。结果:热休克处理上调THP-1巨噬细胞HSP70、TLR4、MCP-1和IL-8的表达。SB203580、anti-TLR4可完全抑制热休克诱导的MCP-1和IL-8表达上调,而PDTC、anti-HSP70对上述效应起部分抑制作用。结论:THP-1细胞经热休克处理后通过HSP70、TLR4/P38MAPK、TLR4/NF-κB途径上调MCP-1和IL-8的表达。 Background:Atherosclerosis(AS) is a chronic inflammatory /autoimmune diseases.A considerable amount of evidence has shown that stress may induce a chronic immune inflammatory process involved in atherosclerosis.But the underlying mechanisms are still largely unknown.Objective:This study intends to research the effects of heat shock on the expression of monocyte chemotactic protein-1(MCP-1) and interleukin-8(IL-8) of THP-1 and the roles of heat shock protein 70(HSP70),toll like receptor 4(TLR4),p38 mitogen-activated protein kinase(P38MAPK) and nuclear factor-kappa B(NF-κ B) during these effects of heat shock.Methods:THP-1 cells were incubated with using 160 nmol/L phorbol myristoyl acetate(PMA) for 24 hs to induce their differentiation into macrophages and then divided into different groups.Group A:common temperature control group.Group B:Heat shock treatment group.THP-1 macrophages were treated 42℃ for 1h,then at 37℃ for 6 hs.Group C:Heat shock + anti-TLR4 antibody group.THP-1 macrophages were added with the 10μg/mL Anti-TLR4 for 2 hs and watered in 42℃ water for 1h,then to 37℃ resumption for 6 hs.Group D:heat shock + P38 MAPK inhibitor SB203580 group.THP-1 macrophages were added with 20μmol/mL SB203580(P38 specially inhibitor) for 30mins and watered in 42℃ water for 1h,then to 37℃ resumption for 6 hs.Group E:heat shock + NF-κ B inhibitor PDTC group.THP-1 macrophages were added with 30μmol/mL PDTC(NF-κ B specially inhibitor) for 30min and watered in 42℃ water for 1h,then to 37℃ resumption for 6 hs.Group F:heat shock + anti-HSP70 antibody group.THP-1 macrophages were added with 2μg/mL anti-HSP70 for 2hs and watered in 42℃ water for 1h,then to 37℃ resumption for 6 hs.The mRNA and protein levels of HSP70,TLR4,MCP-1 and IL-8 in THP-1 cells were tested by reverse transcriptase-polymerase chain reaction(RT-PCR),enzyme-linked immunosorbent assay(ELISA) and western blot.Results:Heat shock increased the expression of TLR4,HSP70,MCP-1 and IL-8 in THP-1 cells.The treatment of THP1 cells with SB203580 and anti-TLR4 inhibited heat shock-induced expression of MCP-1 and IL-8 completely.However,PDTC and anti-HSP70 treatment restrained these effects partially.Conclusion:Heat shock up-regulated the expression of MCP-1 and IL-8 in THP-1 cells through HSP70 /TLR4 /P38MAPK and TLR4 /NF-κB pathway.
出处 《心肺血管病杂志》 CAS 2010年第3期220-224,共5页 Journal of Cardiovascular and Pulmonary Diseases
基金 国家自然科学基金资助课题(No30440065) 北京市自然科学基金资助课题(No7052059)
关键词 热休克蛋白70 TOLL样受体-4 单核细胞趋化蛋白-1 白细胞介素-8 动脉粥样硬化 Atherosclerosis Heat shock protein 70 Toll like receptor 4 Monocyte chemotactic protein-1 Interleukin-8
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参考文献11

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