湖南中医药大学学报2025，Vol.45Issue(12)：2280-2287,8.DOI:10.3969/j.issn.1674-070X.2025.12.006
愈痫灵含药血清抑制小胶质细胞Panx1/P2X7R-NLRP3炎性活化及神经元保护效应研究
Inhibitory effects of Yuxianling medicated serum on microglial Panx1/P2X7R-NLRP3 inflammatory activation and its neuroprotective effects
摘要
Abstract
Objective To investigate the regulatory effects of Yuxianling medicated serum on neuroinflammation in lipopolys-accharide(LPS)-induced BV-2 microglial cells and its protective role against neuronal damage resulting from neuroinflammation.Methods A neuroinflammation model was established using LPS-induced BV-2 microglial cells,with experimental groups including a normal control group,a model group,a blank serum group,and Yuxianling medicated serum groups(5%,10%,and 20%).Cell viability was assessed using the cell counting kit-8(CCK-8)assay.The protein expression levels of Pannexin 1(Panx1),Purinergic P2X Receptor 7(P2X7R),NOD-like receptor family pyrin domain-containing 3(NLRP3),apoptosis-associated speck-like protein containing a card(ASC),cysteinyl aspartate specific proteinase-1(Caspase-1),and ionized calcium-binding adapter molecule 1(Iba-1)were analyzed by Western blot.The expression of Panx1,P2X7R,NLRP3,and Iba-1 proteins were observed using immunofluo-rescence.The concentrations of interleukin-1β(IL-1β),IL-18,and tumour necrosis factor-α(TNF-α)in the cell culture supernatant were measured by ELISA.Subsequently,the culture supernatant from BV-2 microglial cells was collected and added to pure HT22 hippocampal neuronal cells for co-incubation(i.e.,conditioned medium treatment)to investigate the protective effects of Yuxianling medicated serum on HT22 neurons.The survival rate of HT22 cells was determined using the CCK-8 assay.Results Compared to the normal control group,the model group exhibited higher protein expression levels of Panx1,P2X7R,NLRP3,ASC,Caspase-1,and Iba-1(P<0.01);elevated cytokine expressions of IL-1β,IL-18,and TNF-α(P<0.01);reduced hippocampal neuronal cell viability(P<0.01).In comparison with the model group,the 20%concentration Yuxianling medicated serum group showed decreased protein expressions of Panx1,P2X7R,NLRP3,ASC,Caspase-1,and Iba-1(P<0.01);reduced expression levels of IL-1β,IL-18,and TNF-α(P<0.01);and increased viability of hippocampal neuronal cells(P<0.01).Conclusion Yuxianling medicated serum can alleviate neuroinflammation in LPS-induced BV-2 microglial cells,thereby reducing their damaging effects on HT22 neurons.关键词
癫痫/愈痫灵含药血清/脂多糖/BV-2细胞/HT22神经元Key words
epilepsy/Yuxianling medicated serum/lipopolysaccharide/BV-2 cell/HT22 neuron分类
医药卫生引用本文复制引用
张艳菊,潘婷婷,杨辉,张林..愈痫灵含药血清抑制小胶质细胞Panx1/P2X7R-NLRP3炎性活化及神经元保护效应研究[J].湖南中医药大学学报,2025,45(12):2280-2287,8.基金项目
国家自然科学基金项目(82060863) （82060863）
贵州省教育厅青年科技拔尖人才项目(黔教技[2024]331号) （黔教技[2024]331号）
贵州省中医药、民族医药科学技术研究专项课题(QZYY-2024-068) （QZYY-2024-068）
贵州中医药大学校级课题(贵中医科院内[2019]20) （贵中医科院内[2019]20）
贵州中医药大学第二附属医院院内项目(GZEYK[2020]11). （GZEYK[2020]11）
