基于AMPK/PPARα/STAT3途径探讨姜黄素对单核细胞迁移和血管壁巨噬细胞形成的影响  

Effects of Curcumin on Monocyte Migration and Macrophage Formation in Vascular Wall based on AMPK/PPARalpha/STAT3 Pathway

作　　者：陈旭 刘子由[1] 陈洪[1] 胡硕 郭林[1] CHEN Xu;LIU Ziyou;CHEN Hong;HU Shuo;GUO Lin(First Affiliated Hospital of Gannan Medical College,Ganzhou 341000 Jiangxi,China)

机构地区：[1]赣南医学院第一附属医院,江西赣州341000

出　　处：《中药新药与临床药理》2019年第9期1105-1111,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金资助项目（81360287）;江西省自然科学基金资助项目（2010GZY0031）

摘　　要：目的基于腺苷酸活化蛋白激酶/过氧化物酶体增殖物激活受体α/信号传导及转录激活因子3(AMPK/PPARα/STAT3)途径探讨姜黄素对单核细胞迁移和血管壁巨噬细胞形成的影响。方法将人单核细胞株THP-1细胞分为空白对照组、LPS组(lipopolysaccharide,LPS,5μg·mL-1)、LPS+辛伐他汀组(LPS 5μg·mL-1+Simvastatin 5μmol·L-1)、LPS+姜黄素组[用姜黄素(10、20、40、80μmol·L-1)处理2 h后,再用5μg·mL-1LPS刺激THP-1细胞0.5 h];采用Transwell迁移实验检测THP-1细胞迁移能力;采用ELISA法检测高迁移率族蛋白B1(HMGB1)、单核细胞趋化蛋白-1(MCP-1)的蛋白水平;采用RT-PCR法检测MCP-1的mRNA表达;采用Western Blot法检测腺苷酸活化蛋白激酶(AMPK)、过氧化物酶体增殖物激活受体α(PPARα)、信号转导及转录激活因子3(STAT3)的蛋白表达;采用RT-PCR法检测AMPK、PPARα、STAT3的mRNA表达;采用CD68免疫组织化学染色法计算载脂蛋白E(ApoE)小鼠血管壁巨噬细胞的含量。结果与空白对照组相比,LPS组THP-1细胞迁移能力显著升高,HMGB1、MCP-1蛋白及mRNA表达水平显著升高,AMPK、PPARα蛋白及mRNA表达水平均显著降低,STAT3蛋白及mRNA表达水平显著升高(P<0.05或P<0.01);与空白对照组相比,LPS组小鼠血管壁巨噬细胞数量显著升高(P<0.05或P<0.01);与LPS组相比,LPS+姜黄素(10、20、40、80μmol·L-1)组THP-1细胞迁移能力显著降低,HMGB1、MCP-1蛋白及mRNA表达水平显著降低,AMPK、PPARα蛋白及mRNA表达水平均显著升高,STAT3蛋白及mRNA表达水平显著降低(P<0.05或P<0.01)。与LPS组相比,姜黄素各剂量组10、20、40、80 mg·kg-1小鼠血管壁巨噬细胞数量降低(P<0.05或P<0.01);其中高剂量组效果优于低剂量组(P<0.05或P<0.01)。结论姜黄素可抑制LPS诱导的THP-1单核细胞的迁移及血管壁巨噬细胞的形成,其作用机制可能是通过调节AMPK/PPARα/STAT3途径来实现的。Objective To investigate the effects of curcumin(Cur)on monocyte migration and vascular wall macrophage formation based on AMPK/PPARα/STAT3 pathway.Methods Human monocyte THP-1 cells were divided into control group,LPS group(lipopolysaccharide,LPS,5μg?mL-1),LPS+simvastatin group(LPS 5μg·mL-1+simvastatin 5 mol·L-1),LPS+Cur group(treated with Cur(10,20,40,80μmol?L-1)for 2 h,then stimulated with 5μg?mL-1 LPS for 0.5 h)The migration ability of THP-1 cells was detected by Transwell migration,the protein levels of HMGB1 and MCP-1 was detected by ELISA,the mRNA expression of HMGB1 and MCP-1 were detected by RT-PCR.The protein expression of AMPK,PPARαand STAT3 were detected by Western Blot.The mRNA expression of AMPK,PPARαand STAT3 were detected by RT-PCR,and the content of vascular wall macrophages in apolipoprotein E(ApoE)mice was calculated by CD68 immunohistochemical staining.Results Compared with the control group,the migration ability of THP-1 cells,the protein and mRNA expression of HMGB1,MCP-1,and the protein and mRNA expression of AMPK,PPARαwere significantly increased in LPS groups,the protein and mRNA expression of STAT3 increased significantly(P<0.05 or P<0.01).Compared with the control group,the number of murine vascular wall macrophages in LPS group increased significantly(P<0.05 or P<0.01).Compared with LPS group,the migration ability,the protein and mRNA expression of HMGB1,MCP-1 of THP-1 cells were significantly decreased,the protein and mRNA expression of AMPK,PPARαwere significantly increased.The protein and mRNA expression of STAT3 were significantly decreased in the LPS+Cur(10,20,40,80μmol?L-1)group(P<0.05 or P<0.01).Compared with LPS group,the numbers of vascular wall macrophages in Cur(10,20,40,80 mg?kg-1)groups were lower(P<0.05 or P<0.01),and the effect of high dosage group was better than that of low dosage group(P<0.05 or P<0.01).Conclusion Curcumin can inhibit the migration of THP-1 monocytes induced by LPS and the formation of vascular wall macrophages.The mechanism of curcumin ma

关 键 词：AMPK/PPARα/STAT3 姜黄素 单核细胞迁移 血管壁巨噬细胞形成 

分 类 号：R285[医药卫生—中药学]