摘要
目的:探讨鹿血晶(DBC)对小鼠巨噬细胞系RAW 264.7的免疫调节作用及其可能机制。方法:CCK-8法检测细胞活力;流式细胞术及免疫荧光法检测巨噬细胞对大肠杆菌的吞噬能力;qRT-PCR法检测炎症因子iNOS、IL-1β、IL-6的mRNA表达水平;ELISA法检测培养基上清中iNOS、IL-1β、IL-6含量;Western blot法检测转染NF-κB信号通路蛋白表达。结果:鹿血晶能够提高LPS刺激下的RAW 264.7细胞活力(P<0.05);鹿血晶能抑制LPS刺激下RAW 264.7细胞的炎症因子表达和释放(P<0.05);鹿血晶能够抑制LPS刺激下RAW 264.7细胞内磷酸化IKK-α/β和磷酸化P65蛋白的表达;鹿血晶促进RAW 264.7细胞对大肠杆菌的吞噬能力(P<0.05)。结论:鹿血晶能够增强巨噬细胞吞噬大肠杆菌的能力,并且在不影响细胞活力的同时作用于NF-κB信号通路,抑制炎症状态下巨噬细胞炎症因子的表达与释放。
Objective:To investigate the immunoregulating effect of deer blood crystal(DBC)on mouse macrophage line RAW 264.7 and to explore the possible mechanism.Methods:CCK-8 assay was used to detect cell viability.Flow cytometry and immunofluorescence were used to detect the phagocytic index.qRT-PCR assay was used to detect the mRNA level of inflammatory cytokines iNOS,IL-1β,IL-6,ELISA assay was used to detect the contents of iNOS,IL-1β,IL-6 in the supernatant of the culture medium.Western blot was used to detect the expression of protein in the NF-κB signaling pathway.Results:The cell viability of RAW 264.7 cells with LPS stimulation was increased by DBC(P<0.05).The expression and release levels of inflammatory cytokines in DBC+LPS group were significantly lower than those in the LPS group(P<0.05).The expression of phospho-IKK-α/βand phospho-P65 protein in DB+LPS group was lower than that in LPS group.The phagocytic ability of RAW 264.7 cells against E.coli increased after DBC treatment.Conclusion:DBC enhances the ability of macrophages to engulf E.coli,and inhibits macrophages releasing inflammatory factors induced by LPS via NF-κB signaling pathway without affecting cell viability.
作者
李京蔓
潘宇晨
夏晓雨
李丹
窦环(指导)
侯亚义(指导)
LI Jing-Man;PAN Yu-Chen;XIA Xiao-Yu;LI Dan;DOU Huan;HOU Ya-Yi(Nanjing University Medical School,Nanjing 210093,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第7期810-814,共5页
Chinese Journal of Immunology
基金
中央高校基本科研业务费专项资金(021414380342)和江苏省“六大人才高峰”高层次人才项目(YY-021)。
关键词
鹿血晶
巨噬细胞
免疫调节
Deer blood crystal
Macrophages
Immunoregulation
