安徽医科大学学报2025，Vol.60Issue(11)：2059-2068,10.DOI:10.19405/j.cnki.issn1000-1492.2025.11.010
白头翁皂苷B4通过NOS3-DHFR轴对口腔扁平苔藓上皮细胞谷氨酰胺代谢的影响及作用机制
Mechanism of Anemoside B4 on glutamine metabolism in oral lichen planus epithelial cells via the NOS3-DHFR axis
摘要
Abstract
Objective To investigate the mechanism of Anemoside B4(AB4)on glutamine metabolism in oral li-chen planus(OLP)epithelial cells via the nitric oxide synthase 3(NOS3)-dihydrofolate reductase(DHFR)axis.Methods Bioinformatics analysis was performed to identify the intersection of molecular targets of OLP,AB4,and genes related to glutamine metabolism.A lipopolysaccharide(LPS)-induced HOK-16B model of OLP was estab-lished.HOK-16B were divided into Ctrl group,OLP group,AB4 group,OLP+oe-NOS3 group,OLP+sh-NOS3 group,OLP+sh-NOS3+oe-DHFR group,and OLP+sh-NOS3+AB4 group.Cell proliferation was detected by cell counting kit-8(CCK-8);cell apoptosis was detected by TdT-mediated dUTP Nick-End Labeling(TUNEL);inflammatory factors iInterleukin(IL)-1β,tumor necrosis factors-α(TNF-α)concentrations in cell supernatants were measured using enzyme-linked immunosorbent assay(ELISA)kits;glutamine uptake and glutamate produc-tion were determined using kits;and the protein expression of alanine-serine-cysteine transporter2(ASCT2)and glutamine synthase(GLS)was assessed by Western blot.Results Bioinformatics analysis of molecular targets of OLP,AB4,and genes related to glutamine metabolism revealed three intersection targets:NFE2L2,NOS1,and NOS3.Compared with the Ctrl group,the OLP group exhibited decreased HOK-16B cell viability(P＜0.001),increased apoptosis rate(P＜0.01),upregulated concentrations of IL-1 β and TNF-α(P＜0.001),elevated glu-tamine uptake and glutamate production(P＜0.01),and enhanced expression of ASCPT2 and GLS proteins(P＜0.001).Compared with the OLP group,the AB4 group showed improved cell viability(P＜0.05),reduced apop-tosis rate and release of IL-1β and TNF-α(P＜0.05),decreased glutamine uptake and glutamate production(P＜0.05),and downregulated expression of ASCPT2 and GLS proteins(P＜0.001).Compared with the OLP group,the OLP+oe-NOS3 group had increased HOK-16B cell viability(P＜0.01),reduced apoptosis rate(P＜0.05),decreased concentrations of IL-1β and TNF-α(P＜0.05),lowered glutamine uptake and glutamate pro-duction(P＜0.05),and weakened expression of ASCPT2 and GLS proteins(P＜0.01);whereas the OLP+sh-NOS3 group had decreased HOK-16B cell viability(P＜0.05),increased apoptosis rate(P＜0.05),elevated concentrations of IL-1β and TNF-α(P＜0.01),increased glutamine uptake and glutamate production(P＜0.05),and enhanced expression of ASCPT2 and GLS proteins(P＜0.001).Compared with the OLP+sh-NOS3 group,both the OLP+sh-NOS3+oe-DHFR group and the OLP+sh-NOS3+AB4 group showed increased HOK-16B cell viability(P＜0.001),reduced apoptosis rate(P＜0.05),decreased concentrations of IL-1 β and TNF-α(P＜0.01),lowered glutamine uptake and glutamate production(P＜0.05),and weakened expression of AS-CPT2 and GLS proteins(P＜0.05).Conclusion AB4 inhibits the progression of OLP by mediating glutamine metabolism via the regulation of the NOS3-DHFR axis.关键词
白头翁皂苷B4/一氧化氮合酶3/二氢叶酸还原酶/谷氨酰胺代谢/口腔扁平苔藓Key words
Anemoside B4/nitric oxide synthase 3/dihydrofolate reductase/glutamine metabolism/oral lichen planus分类
医药卫生引用本文复制引用
李敏,杨梦华,高毅,张子建,江丹..白头翁皂苷B4通过NOS3-DHFR轴对口腔扁平苔藓上皮细胞谷氨酰胺代谢的影响及作用机制[J].安徽医科大学学报,2025,60(11):2059-2068,10.基金项目
河北省中医药管理局科研计划项目(编号:2021071) Scientific Research Project of Hebei Provincial Administration of Traditional Chinese Medicine(No.2021071) （编号:2021071）
