沉香倍半萜ZH-13抑制JNK磷酸化改善神经小胶质细胞炎症的机制
Sesquiterpene ZH-13 from Aquilariae Lignum Resinatum Improves Neuroinflammation by Regulating JNK Phosphorylation
阴紫钰 1高云 1王俊娇 1薛伟刚 1逄雪萍 1刘慧婷 1赵云芳 2霍会霞 2李军 1郑姣2
作者信息
- 1. 北京中医药大学中医药研究院中药现代研究中心,北京 102488;北京中医药大学中药学院,北京 102488
- 2. 北京中医药大学中医药研究院中药现代研究中心,北京 102488
- 折叠
摘要
目的:研究沉香倍半萜改善神经炎症的药效物质及作用机制.方法:采用脂多糖(LPS)刺激BV-2小胶质细胞,将细胞分为正常组、模型组、ZH-13低、高剂量给药组(10、20 μmol·L-1).模型组给予1 µmol·L-1 LPS.利用细胞增殖与活性检测(CCK-8)试剂盒检测细胞活性,一氧化氮(NO)试剂盒(Griess法)检测细胞上清NO释放.采用实时荧光定量聚合酶链式反应(Real-time PCR)检测细胞中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)、白细胞介素-6(IL-6)mRNA的表达;蛋白免疫印迹法(Western blot)检测丝裂原活化蛋白激酶(MAPK)通路磷酸化激活的变化.利用应激活化蛋白激酶c-Jun氨基末端激酶(JNK)激动剂茴香霉素(Ani)刺激细胞并给药,检测GYF-31对JNK蛋白磷酸化的影响.结果:与模型组比较,ZH-13剂量依赖性地明显降低LPS刺激下BV-2细胞NO释放(P＜0.05,P＜0.01);与模型组比较,20 μmol-L-1剂量ZH-13明显降低细胞中IL-1β、TNF-α、iNOS、IL-6mRNA的表达量(P＜0.05,P＜0.01).与模型组比较,ZH-13低、高剂量给药组炎症因子TNF-α蛋白表达及上游MAPK通路中JNK的磷酸化蛋白表达均明显降低(P＜0.05).在JNK激动剂茴香霉素刺激下,与激动剂组比较,ZH-13低、高剂量给药组能够降低JNK蛋白的磷酸化(P＜0.01).结论:沉香倍半萜类化合物ZH-13可显著改善LPS诱导的BV-2细胞神经炎症反应,通过抑制JNK过度磷酸化激活抑制炎症因子TNF-α的表达,阐明沉香镇静安神的药效物质及作用机制.
Abstract
Objective:To study the pharmacological substances and mechanisms through which sesquiterpene ZH-13 from Aquilariae Lignum Resinatum improves neuroinflammation.Methods:BV-2 microglial cells were stimulated with lipopolysaccharide(LPS)to induce neuroinflammation.The cells were divided into the normal group,the model group,and the ZH-13 low-and high-dose treatment groups(10,20 µmol·L-1).The model group was treated with 1 μmol·L-1 LPS.Cell viability was assessed using the cell proliferation and activity assay(CCK-8 kit).Nitric oxide(NO)release in the cell supernatant was measured using a nitric oxide kit(Griess method).The mRNA expression levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),inducible nitric oxide synthase(iNOS),and interleukin-6(IL-6)were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The phosphorylation of mitogen-activated protein kinase(MAPK)pathway proteins was assessed by Western blot.Results:Compared with the model group,ZH-13 dose-dependently reduced NO release from BV-2 cells under LPS stimulation(P＜0.05,P＜0.01).In the 20 μmol L-1 ZH-13 treatment group,the mRNA expression levels of IL-1β,TNF-α,iNOS,and IL-6 were significantly reduced compared to the model group(P＜0.05,P＜0.01).In both the low-and high-dose ZH-13 groups,the expression of the inflammatory factor TNF-α and the phosphorylation of c-Jun N-terminal kinase(JNK)in the upstream MAPK pathway were significantly reduced(P＜0.05).After stimulation with the JNK agonist anisomycin(Ani),both low-and high-dose ZH-13 treatment groups showed reduced phosphorylation of JNK proteins compared to the Ani-treated group(P＜0.01).Conclusion:The sesquiterpene compound ZH-13 from Aquilariae Lignum Resinatum significantly ameliorates LPS-induced neuroinflammatory responses in BV-2 cells by inhibiting excessive JNK phosphorylation and reducing TNF-α expression.These findings elucidate the pharmacological substances and mechanisms underlying the sedative and calming effects of Aquilariae Lignum Resinatum.
关键词
沉香/倍半萜/小胶质细胞/神经炎症Key words
Aquilariae Lignum Resinatum/sesquiterpene/microglia/neuroinflammation引用本文复制引用
出版年
2025
国家科技期刊平台
NSTL
维普
万方数据