藁本内酯抑制rhHSP60诱导THP-1细胞炎症反应及其机理探讨  

Ligustilide Inhibiting rhHSP60 Inducing THP- 1 Cells Inflammation

作　　者：高颖[1,2] 陈蕊[3] 顾宁[4] 何小丽[4] 

机构地区：[1]南京中医药大学博士后流动站,江苏南京210046 [2]第二军医大学附属公利医院,上海200135 [3]南京中医药大学第一临床医学院,江苏南京210029 [4]南京中医药大学第三附属医院心内科,江苏南京210001

出　　处：《辽宁中医杂志》2016年第9期1926-1930,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81173399);中国博士后科学基金第57批面上项目(2015M571792);江苏省博士后科研资助项目(1401137C)

摘　　要：目的:观察藁本内酯对人重组HSP60诱导THP-1源性巨噬细胞炎症反应的抑制作用,并从TLR4-My D88-NF-κB信号通路角度探讨其抗炎机理。方法:选择人白血病源性单核细胞系THP-1细胞,经200μg·L-1PMA作用24 h,诱导分化为巨噬细胞。设对照组、热休克蛋白60(10μg·mL^(-1))组、藁本内酯(10、20、40μg·mL^(-1))组、辛伐他汀(15μg·mL^(-1))组,加入热休克蛋白60刺激0、6、12、24 h,ELISA法测定以上不同时间点培养液中的TNF-α、IL-6含量;Western blot法测定藁本内酯(10、20、40μg·mL^(-1))预处理24 h、热休克蛋白60(10μg·mL^(-1))刺激12 h后TLR4、My D88及磷酸化NF-κB(p-NF-κB)的蛋白表达。结果:(1)热休克蛋白60组的TNF-α含量在6 h明显升高,与0 h相比,差异有统计学意义(P<0.01),至12 h含量达高峰,随后开始逐渐下降,24 h与12 h相比差异有统计学意义(P<0.01);热休克蛋白60组的IL-6含量自6 h开始逐渐升高,与对照组相比,差异有统计学意义(P<0.01)。(2)与热休克蛋白60组相比,辛伐他汀组的TNF-α含量在6、12、24 h均下降,差异有统计学意义;藁本内酯20、40μg·mL^(-1)组与热休克蛋白60组比较,在以上三个时间点差异均有统计学意义,与辛伐他汀组比较,差异无统计学意义。(3)与热休克蛋白60组相比,辛伐他汀组的IL-6含量在6、12、24 h均下降,差异有统计学意义;藁本内酯20、40μg·mL^(-1)组与热休克蛋白60组比较,在以上三个时间点差异均有统计学意义,与辛伐他汀组比较,差异无统计学意义。(4)藁本内酯20、40μg·mL^(-1)组与热休克蛋白60组相比,TLR4、My D88及p-NF-κB蛋白明显下降,差异有统计学意义。结论:藁本内酯可抑制热休克蛋白60所导致的TNF-α、IL-6等炎症因子升高,具有一定的抗炎作用,该效应与其抑制TLR4-My D88-NF-κB信号通路激活有关。Objective ： To explore the inhibitory effects of ligustilide on the THP - 1 cells induced with human heat shock pro- tein 60（hHSP60） and clarify the underlying mechanism. Methods：The human acute monocytic leukemia cell line THP- 1 ceils were differentiated into macrophages by phorbol - 12 - myristate - 13 - acetate（PMA）. The macrophages were treated with ligusti- lide（ 10,20,40 μg/mL） or simvastatin（ 10 μg/mL） for 24 h followed by recombined hHSP60（ 10 μg/mL） for different times. At the end of the experiments, the contents of TNF - α and IL - 6 in the supernants were measured with ELISA and the protein ex- pressions of TLR4 and MyD88 or phosphorylated NF - KB p65 were measured with western blot. Results ： （ 1 ） The contents of TNF - α and IL -6 were elevated significantly after incubation with rhHSP60 for 6 h. While those in the ligustilide （20,40 μg/mL） or simvastatin pretreated groups were significantly decreased. （2） The protein expressions of TLR4 and MyD88 or phosphorylated NF - KB p65 increased significantly after induced with rhHSP60 which were inhibited by ligustilide （20,40 μg/mL）. Conclusion：Li- gustilide possesses anti - inflammatory properties and can decrease the secretions of TNF - α and IL - 6 induced with rhHSP60 which could be through the inhibition of the activation of the TLRd - MvD88 - NF - KB signaling pathway.

关 键 词：藁本内酯 热休克蛋白60 炎症因子 Toll样受体4 髓样分化因子88 易损斑块 

分 类 号：R285.5[医药卫生—中药学]