解放军医学杂志2025，Vol.50Issue(2)：188-196,9.DOI:10.11855/j.issn.0577-7402.0199.2024.0906
远志皂苷元缓解脂多糖诱导的小鼠小胶质细胞炎症反应的机制
Mechanism of senegenin in improving lipopolysacchride-induced inflammatory response of BV2 microglial cell
摘要
Abstract
Objective To investigate the mechanism by which Senegenin(SEN)alleviates microglial inflammatory response through the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)pathway.Methods BV2 mouse microglia cells were randomly divided into control group,model group,SEN group and MCC950 group.Cells in control group were not treated,and cells in model group were added with 1 μg/ml lipopolysaccharide(LPS);Cells in SEN group were added with 1 μg/ml LPS+4 μmol/L SEN,and cells in MCC950 group were added with 1 μg/ml LPS+10 μmol/L MCC950 for 24 hours.CCK-8 method was used to detect the effect of different concentrations of SEN on the viability of BV2 cells.Griess method was used to determine the release amount of nitric oxide(NO)in the supernatant.Real-time fluorescent quantitative PCR was used to determine the mRNA expression levels of NLRP3,lymphocyte apoptosis-associated spect-like protein containing a CARD(ASC),caspase-1,interleukin(IL)-1β and IL-18 mRNA.Immunofluorescence staining was used to detect the expression levels of ASC,IL-1β,Nrf2 and heme oxygenase-1(HO-1).Western blotting was used to detect the expression levels of NLRP3,caspase-1,ASC,IL-1β,IL-18,Nrf2,HO-1,nuclear factor kappa B(NF-κB)and inducible nitric oxide synthase(iNOS).Results The results of CCK-8 method showed that there was no significant difference in the viability of BV2 cells treated with 2～20 μmol/L SEN compared with control group(P>0.05).Compared with control group,the viability of BV2 cells in model group decreased significantly(P<0.05).Compared with model group,the viability of BV2 cells in 4 μmol/L SEN group was significantly restored(P<0.05).Compared with control group,the results of Griess method showed that the release amount of NO in cells of model group increased significantly(P<0.05);the results of real-time PCR showed that the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA in cells of model group increased significantly(P<0.05);the results of Western blotting showed that the protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 proteins in cells of model group increased significantly(P<0.05),and the immunofluorescence staining results showed that the expression levels of iNOS and NF-κB protein in cells of model group increased,and the expression levels of Nrf2 and HO-1 decreased,with statistically significant differences(P<0.05).Compared with model group,the release amount of NO in cells of SEN group and MCC950 group decreased,and the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA and proteins decreased,with statistically significant differences(P<0.05);in the SEN group,the expression levels of iNOS and NF-κB decreased,and immunofluorescence staining showed that Nrf2 was translocated into the nucleus,and the expression levels of Nrf2 and HO-1 proteins increased significantly,with statistically significant differences(P<0.05).Conclusions SEN could alleviate the inflammatory response of mouse microglia cells induced by LPS and inhibit the activation and expression of NLRP3 inflammasome,with an effect comparable to that of the inflammasome inhibitor MCC950.The mechanism may be related to the regulation of the expression of upstream factors Nrf2 and HO-1.关键词
远志皂苷元/脂多糖/小胶质细胞/NLRP3炎性小体/炎症反应Key words
senegenin/lipopolysaccharides/BV2 cells/NLRP3 inflammasome/inflammatory response分类
临床医学引用本文复制引用
穆秉桃,尉杰忠,宋丽娟,郭敏芳,于婧文,曹佳蕾,杨凤君,贾思玮,苏琴,孟涛,马存根..远志皂苷元缓解脂多糖诱导的小鼠小胶质细胞炎症反应的机制[J].解放军医学杂志,2025,50(2):188-196,9.基金项目
国家自然科学基金(82004028) （82004028）
山西省中医重点实验室建设项目(zyyyjs2024027) （zyyyjs2024027）
山西省"四个一批"科技兴医创新计划项目(2022XM33) （2022XM33）
山西省卫健委中医药科研课题立项计划(2022ZYYC090) （2022ZYYC090）
山西省中医药管理局科研课题(2023ZYYB2042) This work was supported by the National Natural Science Foundation of China(82004028),the Project of Key Research Laboratory of Traditional Chinese Medicine of Shanxi Province(zyyyjs2024027),the"Four Batches"Science and Technology Innovation Plan Project of Shanxi Province(2022XM33),the Chinese Medicine Scientific Research Project Establishment Plan of Shanxi Provincial Health Commission(2022ZYYC090),and the Traditional Chinese Medicine Research Project of Shanxi Province(2023ZYYB2042) （2023ZYYB2042）
