ERK通路在凋亡细胞抑制CCL5表达中作用和机制的研究

doi:10.3969/j.issn.10073205.2018.11.002

河北医科大学学报

ERK通路在凋亡细胞抑制CCL5表达中作用和机制的研究

1.河北省石家庄市第一医院感染管理控制处，河北石家庄 050011;2.河北医科大学公共卫生学院预防医学2014级，
河北石家庄 050017;3.河北医科大学基础医学院临床医学2014级，河北石家庄 050017;
4.河北医科大学基础医学院免疫学教研室，河北石家庄 050017

出版日期:2018-11-25 发布日期:2018-11-21
作者简介:仝宇红(1970-),女,河北无极人, 河北省石家庄市第一医院副主任医师，医学博士,从事医院感染研究。
基金资助:
河北省留学人员科技活动资助项目（2014003031）；河北省科技计划项目（17397744D）

The role and mechanism of ERK pathway in apoptotic cells inhibiting CCL5 expression

1.Department of Nosocomial Infection Management, the First Hospital of Shijiazhuang, Hebei Province,
Shijiazhuang 050011, China; 2.Preventive Medicine Grade 2014, School of Public Health, Hebei Medical
University, Shijiazhuang 050017, China; 3.Clinical Medicine Grade 2014, School of Basic Medical
Sciences, Hebei Medical University, Shijiazhuang 050017, China; 4.Department of Immunology,
School of Basic Medical Sciences, Hebei Medical University, Shijiazhuang 050017, China

Online:2018-11-25 Published:2018-11-21

摘要/Abstract

摘要： ［摘要］〓
〖HTH〗目的〖HTSS〗〖KG*2〗探讨细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)通路对凋亡细胞抑制脂多糖(lipopolysaccharides，LPS)诱导巨噬细胞分泌炎症性趋化因子5(CC cehemokine ligand 5,CCL5)的影响。
〖HTH〗方法〖HTSS〗〖KG*2〗用Jurakt细胞制备凋亡细胞,并用流式细胞仪检测细胞凋亡率。用LPS刺激骨髓诱导巨噬细胞(bone marrow induced macrophages，BMDM)和RAW264.7细胞，观察凋亡细胞对LPS诱导的CCL5表达的影响。观察ERK通路在凋亡细胞抑制CCL5表达中的作用，用对照质粒PcDNA3.1、MEKK2/3和MEK5质粒转染RAW264.7细胞，分别以LPS（1 mg/L）、凋亡细胞（apo）和LPS（1mg/L）+apo作用RAW细胞6 h，收集细胞，实时定量PCR技术检测CCL5 mRNA表达。阻断ERK通路，观察CCL5 mRNA表达量的变化。
〖HTH〗结果〖HTSS〗〖KG*2〗LPS刺激组和LPS+凋亡细胞刺激组CCL5 mRNA表达明显高于空白组和凋亡细胞刺激组，LPS+凋亡细胞刺激组CCL5 mRNA表达明显低于LPS刺激组（P＜005）。PD98059 5 mmoL/L、10 mmoL/L、20 mmoL/L组mCCL5 mRNA表达低于DMSO组，PD98059 10 mmoL/L、20 mmoL/L组mCCL5 RNA表达低于PD98059 5 mmoL/L组，PD98059 20 mmoL/L组CCL5 mRNA表达低于PD98059 10 mmoL/L组（P＜005）。PcDNA3.1转染组、MEKK2/3转染组和MEK5转染组CCL5 mRNA水平明显高于对照组，MEKK2/3转染组和MEK5转染组CCL5 mRNA水平明显高于PcDNA3.1转染组，MEK5转染组CCL5 mRNA水平明显高于MEKK2/3转染组（P＜005）。D98059组CCL5 mRNA衰变速度快于空白组（P＜005）。
〖HTH〗结论〖HTSS〗〖KG*2〗凋亡细胞能够抑制LPS诱导的CCL5表达，ERK通路在凋亡细胞抑制CCL5表达中发挥重要作用。

关键词: 炎症趋化因子类, 脂多糖, 细胞外调节蛋白激酶通路

Abstract: ［Abstract］〓Objective〖HTSS〗〓To investigate the effect of extracellular regulated protein kinases(ERK) pathway on apoptotic cells inhibiting lipopolysaccharides(LPS) induced inflammatory CC chemokine ligand 5(CCL5) secreted by macrophages.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓Apoptotic cells were prepared by Jurakt cells and the apoptosis rate was detected by flow cytometry. Bone marrow induced macrophages(BMDM) and RAW264.7 cells were stimulated by LPS to observe the effect of apoptotic cells on the expression of CCL5 induced by LPS. To observe the role of ERK pathway in inhibiting CCL5 expression in apoptotic cells, RAW264.7 cells were transfected with control plasmids PcDNA3.1, MEKK2/3 and MEK5. The cells were treated with LPS(1 mg/L), apoptotic cells(apo) and LPS(1 mg/L) + apo for 6 hours respectively. The expression of CCL5 mRNA was detected by realtime quantitative PCR. The ERK pathway was blocked and the expression of CCL5 mRNA was observed.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓The expression of CCL5 mRNA in LPS stimulation group and LPS + apoptotic cell stimulation group was significantly higher than that in blank group and apoptotic cell stimulation group. The expression of CCL5 mRNA in LPS + apoptotic cell stimulation group was significantly lower than that in LPS stimulation group(P＜0.05). The expression of CCL5 mRNA in PD98059 5 mmoL/L, 10 mmoL/L, 20 mmoL/L group was lower than that in DMSO group, the expression of CCL5 mRNA in PD98059 10 mmoL/L, 20 mmoL/L group was lower than that in PD98059 5 mmoL/L group, and the expression of CCL5 mRNA in PD98059 20 mmoL/L group was lower than that in PD98059 10 mmoL/L group(P＜005). CCL5 mRNA levels in PcDNA3.1 transfection group, MEKK2/3 transfection group and MEK5 transfection group were significantly higher than those in control group, MEKK2/3 transfection group and MEK5 transfection group were significantly higher than those in PcDNA3.1 transfection group, and CCL5 mRNA levels in MEK5 transfection group were significantly higher than those in MEKK2/3 transfection group(P＜005). The decay rate of CCL5 mRNA in group D98059 was faster than that in blank group(P＜005).
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓Apoptotic cells can inhibit the expression of CCL5 induced by LPS, and ERK signaling pathway plays an important role in the inhibition of CCL5 expression by apoptotic cells.

Key words: chemokines, lipopolysaccharides, extracellular regulated protein kinases pathway

仝宇红1，杨〓茜2，尹〓默2，沈佳程3，王恩平2，钱雪松4*. ERK通路在凋亡细胞抑制CCL5表达中作用和机制的研究[J]. 河北医科大学学报, doi: 10.3969/j.issn.10073205.2018.11.002.

TONG Yuhong1, YANG Xi2, YIN Mo2, SHEN Jiacheng3, WANG Enping2, QIAN Xuesong4*. The role and mechanism of ERK pathway in apoptotic cells inhibiting CCL5 expression[J]. Journal of Hebei Medical University, doi: 10.3969/j.issn.10073205.2018.11.002.

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ERK通路在凋亡细胞抑制CCL5表达中作用和机制的研究

The role and mechanism of ERK pathway in apoptotic cells inhibiting CCL5 expression

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