瘦素对单核细胞THP1分泌趋化因子的影响及其作用机制  

Influence of leptin on secretion of chemokine in THP1 cells and its mechanism

作　　者：曹红[1] 王林[1] 李矿发[1] 庞雪利[1] 苏敏[1] 黄云秀[1] 魏兰[1] 陈婷梅[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《吉林大学学报（医学版）》2014年第3期482-487,I0001,共7页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金面上项目资助课题(81272544);重庆市科委自然科学基金计划项目资助课题(cstc2012jjA10011)

摘　　要：目的:探讨瘦素促进单核细胞THP1分泌趋化因子的作用及其相关机制,为研究瘦素调节机体免疫功能提供依据。方法:采用RT-PCR法和流式细胞术检测THP1细胞瘦素受体(Ob-Rb和Ob-Rt)表达。选取对数生长期THP1细胞,随机分为空白对照组和10、50、100及200μg·L-1瘦素组,采用Transwell实验检测各处理组穿膜细胞数。THP1细胞分为空白对照组和100μg·L-1瘦素组,采用Western blotting法检测信号通路关键分子p-AKT、p-ERK 1/2和p-STAT3表达水平。THP1细胞分为空白对照组、100μg·L-1瘦素组、100μg·L-1瘦素+DMSO组、100μg·L-1瘦素+50μmol·L-1 AG490组、100μg·L-1瘦素+10μmol·L-1LY294002组和100μg·L-1瘦素+10mol·L-1PD980590组,采用RT-PCR法和Western blotting法检测各组细胞因子IL-8表达水平。结果:瘦素长受体Ob-Rb和短受体Ob-Rt在THP1细胞中均有高表达。与空白对照组比较,50、100和200μg·L-1瘦素组穿膜细胞数明显增加(P<0.05)。与空白对照组比较,100μg·L-1瘦素组THP1细胞中p-AKT、p-ERK 1/2和p-STAT3表达水平明显升高(P<0.05)。与空白对照组比较,50、100和200μg·L-1瘦素组THP1细胞中IL-8表达水平明显升高(P<0.05);与100μg·L-1瘦素组比较,100μg·L-1瘦素+10μmol·L-1 LY294002组和100μg·L-1瘦素+10mol·L-1 PD980590组THP1细胞中IL-8表达水平明显降低(P<0.05),而100μg·L-1瘦素+50μmol·L-1 AG490组IL-8表达水平变化不明显(P>0.05)。结论:瘦素能促进单核细胞THP1分泌趋化因子,其机制可能与PI3K/AKT和MAPK/ERK 1/2信号通路有关。Objective To investigate the effect of leptin on the secretion of chemokine in THP1 cells and explore its related mechanism, and to provide basis for research on the role of leptin in immune response.Methods The expressions of Ob-Rb and Ob-Rt in THP1 cells were detected by RT-PCR and flow cytometry （FCM）.The THP1 cells at logarithm growth phase were selected and randomly divided into blank control group and different concentrations（10,50,100,200μg· L-1 ）of leptin groups.Transwell chamber assay was performed to detect the number of invated THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group.Western blotting method was carried out to detect the expressions of p-AKT,p-ERK 1/2,and p-STAT3 in THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group,100μg·L-1＆amp;nbsp;leptin＋ DMSO group,100μg·L-1 leptin＋50μmol·L-1 AG490 group,100μg·L-1 leptin＋10μmol·L-1 LY294002 group and 100μg·L-1 leptin＋ 10 mol·L-1 PD980590 group.RT-PCR and Western blotting methods were performed to detect the expression of IL-8.Results Ob-Rb and Ob-Rt were highly expressed in THP1 cells. Compared with blank control group,the number of invated THP1 cells was significantly increased in 50,100,and 200μg·L-1 leptin groups （P〈0.05）.Compared with blank control group,the expressions of p-STAT3,p-ERK 1/2 and p-AKT in THP1 cells were up-regulated in 100 ug·L-1 leptin group（P〈0.05）.Compared with blank control group,the mRNA and protein expressions of IL8 in THP1 cells in 50,100,and 200μg·L-1 leptin groups were remarkably increased（P〈0.05）;compared with 100μg·L-1 leptin group,the expressions of IL-8 in THP1 cells in 100μg·L-1 leptin＋10 mol·L-1 PD980590 group and 100μg·L-1 leptin＋10μmol·L-1 LY2 94002 group were decreased（P〈0.05）,while the expression of IL-8 in 100μg·L-1 leptin＋ 50μmol·L-1 AG490 group had no change（P〉0.05）.Conclusion leptin can up-regulate the expression of chemokine in THP1 cells,whi

关 键 词：瘦素 THP1 细胞 趋化因子 白细胞介素-8 

分 类 号：R392.12[医药卫生—免疫学]