摘要
目的探讨miR-142-3p对人单核细胞株THP-1释放炎症因子的调控作用。方法体外培养人单核细胞株THP-1,(1)以不同浓度(0.5和2.0μg/mL)脂多糖(LPS)诱导THP-1细胞24 h;(2)将miR-142-3p的拟似物(mimic,100 nmol/L)转染至THP-1细胞48 h;(3)先将miR-142-3p的抑制剂(inhibitor,100 nmol/L)转染至THP-1细胞24 h,再用不同浓度(0.5和2.0μg/mL)LPS诱导THP-1细胞24 h。处理后收集细胞培养液,用ELISA法分别检测THP-1细胞培养液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和单核细胞趋化蛋白-1(MCP-1)的释放量。结果 (1)两种浓度LPS诱导THP-1细胞后,TNF-α、IL-6和MCP-1的释放量均增加(P<0.01),并表现出LPS浓度依赖性,高浓度者释放量较低浓度者增加。(2)THP-1细胞转染miR-142-3p mimic 48 h后,TNF-α、IL-6和MCP-1的释放量增加(P<0.05,P<0.01)。(3)THP-1细胞转染miR-142-3p inhibitor 48 h后,再以0.5μg/mL LPS诱导后,IL-6的释放量减少(P<0.05)。结论 LPS能诱导THP-1细胞释放TNF-α、IL-6和MCP-1。过表达miR-142-3p能促进THP-1细胞释放TNF-α、IL-6和MCP-1。抑制THP-1细胞miR-142-3p的表达可减少LPS诱导的IL-6释放。miR-142-3p参与调控THP-1的炎症因子释放,提示miR-142-3p在调控免疫细胞的炎症反应过程中具有重要的作用。
Objective To investigate the role of miR - 142 -3p on inflammatory factors released from THP - 1 cells. Methods Human monocytes THP - 1 cells were cultured in vitro. （ 1 ） THP - 1 cells were cultured for 24 h with 0. 5 μg/mL and 2. 0 μg/mL lipopolysaccharides （LPS）. （2）THP - 1 cells were transfected with miR - 142 -3p mimic （100 nmol/L） by lipofectamine RNAi MAX for 48 h. （3）THP- 1 cells were transfected with miR- 142 -3p inhibitor （ 100 nmoL/L） by lipofectamine RNAi MAX for 24 h, and cultured by 0. 5 μg/mL and 2. 0 μg/mL LPS for 24 h. The concentrations of TNF - a, IL - 6 and MCP - 1 in the cultured supernatant were assessed by ELISA. Results （ 1 ） The concentrations of TNF - a, IL - 6 and MCP - 1 in THP - 1 cells stimulated with LPS were significantly increased with dose- dependent manners. （2） The transfeetion of miR- 142 -3p mimic significantly increased the release of TNF-a, IL- 6 and MCP- 1. （3） However, the pre- treatment of miR- 142 -3p inhibitor reversed the LPS- induced release of IL - 6 from THP - 1 cells. Conclusion miR - 142 - 3p may play an important role in the regulation on inflammatory fac- tors released from THP - 1 cells.
出处
《广东医学》
CAS
CSCD
北大核心
2014年第2期180-183,共4页
Guangdong Medical Journal
基金
广东省自然科学基金资助项目(编号:10151018201000018
S2013010014887)
广东省科技计划项目(编号:2012B031800233)
