摘要
目的观察芹菜素对脂多糖(LPS)诱导小鼠海马神经元细胞损伤的改善作用,并探讨其相关机制。方法体外培养小鼠海马神经元细胞系HT22,采用CCK-8法筛选造模药物LPS及干预药物芹菜素的最佳作用浓度,筛选得到LPS、芹菜素的最佳作用浓度分别为2µg/mL、0.25µmol/L。将HT22细胞分为对照组、模型组及芹菜素组,对照组正常培养不作任何处理,模型组加入0.25µmol/L的LPS培养,芹菜素组以2µg/mL的LPS干预24 h后,再加入0.25µmol/L芹菜素培养。采用Annexin V/PI双染法观察细胞凋亡情况,Western blotting法检测细胞肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)蛋白。采用UPLC-Q-TOF-MS/MS分析并筛选各组细胞差异代谢产物,对差异代谢产物进行MetPA分析,获取其潜在的代谢途径。结果细胞凋亡率及TNF-α、IL-1β蛋白表达模型组>芹菜素>对照组(P均<0.01)。代谢组学分析共筛选出尿苷二磷酸葡萄糖、谷氨酸、琥珀酸、柠檬酸、左旋肉碱等16个差异代谢产物,共涉及亚油酸、丙氨酸、天冬氨酸和谷氨酸代谢、三羧酸循环、谷胱甘肽代谢等28条代谢途径。结论芹菜素对LPS诱导的小鼠海马神经元细胞损伤具有改善作用,其机制可能与调控细胞内代谢紊乱有关。
Objective To observe the ameliorative effect of apigenin on lipopolysaccharide(LPS)-induced injury of hippocampal neuron cells in mice,and to explore its related mechanism.Methods Mouse hippocampal neuron cell line HT22 was cultured in vitro,and the optimal concentrations of LPS and apigenin were screened by CCK-8 method.The optimal concentrations of LPS and apigenin were 2μg/mL and 0.25μmol/L,respectively.HT22 cells were divided into the control group,model group and apigenin group.Cells in the control group were cultured normally without any treatment,cells in the model group were cultured by adding 0.25μmol/L LPS,and cells in the apigenin group were cultured by intervening with 2μg/mL of LPS for 24 h and then adding 0.25μmol/L apigenin.The apoptosis was observed by Annexin V/PI double staining,and the cellular tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)proteins were detected by Western blotting.UPLC-Q-TOF-MS/MS was used to analyse and screen the differential metabolites of the cells in each group,and the differential metabolites were subjected to MetPA analysis to obtain their potential metabolic pathways.Re⁃sults The apoptosis rates and the expression levels of TNF-αand IL-1βproteins were as follows:the model group>the apigenin group>the control group(all P<0.01).Metabolomic analysis screened out 16 different metabolites,such as uridine diphosphate glucose,glutamic acid,succinic acid,citric acid and L-carnitine,which involved 28 metabolic pathways such as linolic acid,alanine,aspartic acid and glutamic acid metabolism,TCA cycle,and glutathione metabolism.Conclusion Apigenin can ameliorate LPS-induced mouse hippocampal neuronal cell injury,and its mechanism may be related to the regulation of intracellular metabolic disorders.
作者
杨萌
薛傲
李雪
赵红梅
任毅帆
雷霞
张亚峰
张宁
YANG Meng;XUE Ao;LI Xue;ZHAO Hongmei;REN Yifan;LEI Xia;ZHANG Yafeng;ZHANG Ning(College of Pharmacy,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,China;不详)
出处
《山东医药》
CAS
2024年第29期15-19,共5页
Shandong Medical Journal
基金
国家自然科学基金面上项目(82174007)
中央支持地方高校改革发展资金人才培养项目
江苏省中医退行性骨关节病临床医学创新中心资助项目(20210405)。
