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|本期目录/Table of Contents|

PTEN对ox-LDL诱导的巨噬细胞炎性因子的影响及机制研究

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2014年第6期
页码:
646-651
栏目:
基础研究
出版日期:
2014-06-25

文章信息/Info

Title:
Effect of phosphatase and tensin homolog deleted on chromosome ten on ox-LDL-induced inflammatory cytokine secretion from macrophages and its underlying mechanism
作者:
吴晓鹏1; 王选琦1; 李 妍2; 李伟杰2
(1.陕西省第四人民医院心血管内科，陕西 西安 710043；
2.第四军医大学西京医院心血管内科，陕西 西安 710032)
Author(s):
WU Xiao-peng1;  WANG Xuan-qi1;  LI Yan2;  LI Wei-jie2
(1.Department of Cardiology, Shaanxi Fourth People’s Hospital, Xi’an 710043, Shaanxi, China;
2.Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China)
关键词:
PTEN; 巨噬细胞; 炎症; PI3K/AKt; TLR4-NF-κB
Keywords:
PTEN;  macrophage;  inflammatory;  PI3K/AKt;  TLR4-NF-κB
分类号:
R363
DOI:
-
文献标识码:
A
摘要:
目的:分析第10号染色体缺失的磷酸酶和张力蛋白同源基因（PTEN）对氧化低密度脂蛋白(ox-LDL)诱导的巨噬细胞炎症因子影响及其作用机制。方法： 将构建pcDNA3.1(+)-PTEN（rPTEN）重组表达载体及PTEN siRNA转染小鼠巨噬细胞系RAW 264.7，用制备的50 mg/L的ox-LDL孵育24 h，RT-PCR及Western blot分析PTEN的mRNA及蛋白表达水平；ELISA检测炎性因子TNF-α和IL-6的水平；同时Western blot分析对Toll样受体4(TLR4)及转录因子-κB (NF-κB)的影响及其作用机制。结果： PTEN过表达增高了ox-LDL诱导的TNF-α和IL-6的水平； PTEN沉默抑制了ox-LDL诱导的TNF-α和IL-6炎性因子水平。进一步分析表明，PTEN过表达加强了巨噬细胞中ox-LDL诱导的TLR4及其下游NF-κB通路的活化，而抑制其表达后，TLR4-NF-κB通路明显受到抑制；用TLR4特异性抗体预处理后，PTEN过表达诱导的TNF-α和IL-6的水平明显下降。进一步机制分析证实，磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)抑制剂LY294002（1 μmol/L）预处理后，PTEN沉默抑制的TLR4-NF-κB通路的活性明显增加，且伴随有TNF-α和IL-6水平的上调。结论： PTEN有可能通过负向调节PI3K/AKT抗炎通路来影响TLR4-NF-κB炎性通路，进而参与巨噬细胞介导的炎症进程。因此，本研究将为心脑血管疾病的防治提供新的靶标。
Abstract:
AIM:To investigate the effect of phosphatase and tensin homolog deleted on chromosome ten (PTEN) on ox-LDL-induced inflammatory cytokine levels in macrophages and its underlying mechanism. METHODS: The constructed pcDNA3.1(+)-PTEN (rPTEN) recombinant vector and PTEN siRNA were transfected into RAW 264.7 followed by stimulation with 50 mg/L ox-LDL for 24 h. Real-time PCR (RT-PCR) and Western blotting were used to analyze the expression levels of PTEN mRNA and protein. ELISA was used to detect the inflammatory cytokine levels of TNF-α and IL-6. The expression level of TLR4-NF-κB and its underlying mechanism were also explored. RESULTS: Overexpression of PTEN enhanced ox-LDL-induced inflammatory cytokine levels of TNF-α and IL-6, whereas its silencing attenuated this progress. PTEN upregulation increased the expression of ox-LDL-induced TLR4 and its downstream p65 NF-κB. However, silencing PTEN expression with PTEN siRNA significantly decreased ox-LDL-triggered activation of TLR4-NF-κB signaling. Further mechanism analysis demonstrated that preconditioning with PI3K/AKt specific inhibitor LY294002 (1 μmol/L) dramatically augmented PTEN silencing-decreased activation of TLR4-NF-κB pathway, accompanied by the increase in TNF-α and IL-6 levels. CONCLUSION: PTEN may regulate macrophage inflammatory by PI3K/AKt-TLR4-NF-κB pathway. This study thus provides a potential target for the treatment of cardiovascular diseases.

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备注/Memo

备注/Memo:
收稿日期：2014-03-27.
基金项目：国家自然科学基金项目资助（81170184）
作者简介：吴晓鹏，主治医师,硕士生Email：submission029@gmail.com
更新日期/Last Update: 2014-07-10