Human placental extracts negatively regulate LPS-induced microglial inflammatory responses through TLR4/NF-κB signaling pathway
10.3969/j.issn.1000-484X.2025.10.013
- VernacularTitle:人胎盘提取物通过TLR4/NF-κB信号通路负调控LPS诱导的小胶质细胞炎症反应
- Author:
Jingjing HE
1
;
Tongqian WU
;
Shirong YAN
;
Shihui MO
;
Jing LI
;
Fang YU
Author Information
1. 中山大学附属第一医院贵州医院检验科,贵阳 550031
- Publication Type:Journal Article
- Keywords:
Human placental extracts;
Lipopolysaccharide;
Microglia;
Inflammation;
TLR4/NF-κB signaling pathway
- From:
Chinese Journal of Immunology
2025;41(10):2380-2385,2391
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the regulation and mechanism of human placental extract(HPE)on lipopolysaccharide(LPS)-induced BV2 microglial cell inflammation.Methods:Microglia cell lines(BV2)were cultured in vitro and divided into PBS group,HPE group,LPS group and LPS+HPE group.BV2 cell viability was measured by CCK-8 analysis.A fluorescent probe targeting reactive oxygen species(ROS)was to detect the level of intracellular ROS.The mRNA levels of TNF-α,IL-1β and IL-6 were detected by RT-qPCR.The supernatants of different treatment groups were collected.The content of nitric oxide(NO)was detected by the Griess method,and the secretion levels of TNF-α,IL-1β and IL-6 were detected by the flow magnetic bead microarray(CBA)method.The indirect contact co-culture system between BV2 and mouse hippocampal neurons cell line HT22 cells was established to evaluate the neurotoxicity of HPE by the assessment of the cell viability and apoptosis of HT22 cells using CCK-8 or flow cytometry.The potential signaling molecules of NF-κB signaling pathway was detected by flow cytometry.Results:Compared with the PBS group,1.2 μg/ml LPS and 50 ng/ml HPE significantly inhibited the activity of BV2 cells.Compared with the PBS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells of the LPS group were significantly increased(P<0.05),and the secretion levels of NO,TNF-α,IL-1βand IL-6 in the supernatant were also significantly upregulated(P<0.05).The expressions of related signaling pathway molecules Toll-like receptor 4(TLR4),pIκBα and pNF-κB p65 were significantly increased(P<0.05).Compared with the LPS group,the mRNA levels of TNF-α,IL-1β and IL-6 in BV2 cells,the secretion levels of NO,TNF-α,IL-1β and IL-6,the neurotoxicity and neuronal apoptosis induced by microglial conditional medium,and the expressions of TLR4,pIκBα and pNF-κB p65 in the LPS+HPE group were significantly downregulate(P<0.05).Conclusion:HPE may alleviate the microglial inflammation,possibly through the TLR4/NF-κB signaling pathway.
