钩藤中钩藤碱对肿瘤坏死因子-α诱导人星型胶质细胞炎症模型作用机制研究

摘要:

目的研究钩藤中钩藤碱对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导人星型胶质细胞(Human Astrocytes,HA)的炎症模型的作用机制.方法:体外培养HA,用TNF-α诱导HA造模,将细胞分为正常组、模型组、钩藤碱组、依达拉奉组.采用噻唑蓝(MTT)法检测钩藤碱对HA增殖能力的影响,用逆转录PCR法(RT-PCR)检测一氧化氮合酶(nitric oxide synthase,NOS)mRNA、白细胞介素-23(interleukin-23,IL-23)mRNA、白细胞介素-6(interleukin-6,IL-6)mRNA的表达,用分光光度法检测一氧化氮(nitric oxide,NO)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)的表达,用蛋白免疫印迹法(Western blot)检测p-mTOR,mTOR,p-Stat3和Stat3的蛋白表达.结果与正常组比较,32ng·mL-1的TNF-α对星形胶质细胞有明显抑制作用(P<0.001);400μmol·L-1的钩藤碱对星形胶质细胞活力有明显抑制作用(P<0.001).与模型组相比,200μmol·L-1的钩藤碱对模型细胞有明显的增殖作用(P<0.01);钩藤碱组和依达拉奉组NOS mRNA,NO和SOD都显著上升(P<0.001),IL-23mRNA,IL-6mRNA和MDA都显著下降(P<0.001);钩藤碱组p-mTOR/mTOR和p-Stat3/Stat3都显著下降(P<0.01).结论钩藤有效成分钩藤碱具有抑制TNF-α诱导HA炎症的损伤作用,可能是通过下调mTOR/STAT3信号通路增加氧化相关因子NOS,NO,SOD的表达,减少氧化相关因子MDA以及炎症相关因子IL-23和IL-6的表达.

Objective This study aims to study the mechanism of rhynchophylline in the inflammation model of human astrocyte(HA)induced by tumor necrosis factor-α(TNF-α).Method HA was cultured in vitro,was induced by TNF-α,and the cells were divided into normal group,model group,rhynchophylline group and edaravone group.The effect of rhynchophylline on the proliferative capacity of HA was detected by MTT assay.Nitric oxide synthase(NOS)mRNA and interleukin-23(IL-23)mRNA,interleukin-6(IL-6)mRNA expression were detected by reverse transcription-PCR(RT-PCR).Determination of nitric oxide(NO),superoxide dismutase(SOD)and malondialdehyde(MDA)by spectrophotometry.The protein expression of p-mTOR,mTOR,p-Stat3 and Stat3 were detected by Western blot.Result Compared with the normal group,32 ng·mL-1 TNF-αsignificantly inhibited astrocytes viability(P<0.001)and 400μmol·L-1 rhynchophylline significantly inhibited astrocyte viability(P<0.001).Compared with the model group,200μmol·L-1 of rhynchophylline had significant proliferation effects on model cells(P<0.01);NOS mRNA,NO and SOD in the rhyndamine group and captopril group were significantly increased(P<0.001);IL-23 mRNA,IL-6 mRNA and MDA in the rhyndamine group and edaravone group were significantly decreased(P<0.001);p-mTOR/mTOR and p-Stat3/Stat3 were significantly decreased in the rhyndamine group(P<0.01)Conclusion:The active constituent of Uncaria sinensis,rhynchophylline,has the inhibitory effect of inhibiting inflammation of TNF-α-induced HA,possibly by down-regulating mTOR/STAT3 signaling pathway to increse the expression of NOS,NO and SOD in oxidative related factors,reduce the expression of MDA in oxidative related factors and IL-23 and IL-6 in inflammation-related factors.

作者:

安畅 张颖 马阮昕 秦劭晨

An Chang;Zhang Ying;Ma Ruanxin;Qin Shaochen(Guangzhou Yuexiu District Guangta Street Community Health Service Center, Guangzhou 510120,China;Department of Pharmacy,The Sixth Affilited Hospital of SUN Yat-sen University, Guangzhou 510655,China;The Hospital of Shanxi College of Traditional Chinese Medicine,Taiyuan 030012,China)

机构地区:

广州市越秀区光塔街社区卫生服务中心 中山大学附属第六医院药学部 山西中医学院附属医院

出处:

《betway官方app 学报：自然科学版》 CAS 北大核心 2019年第2期98-104,共7页

基金:

国家自然科学基金(81804142) 广东省自然基金项目(2018A030307037) 广东省中医药管理局项目(20181114) 山西省卫生和计划生育委员会科研项目(201601119)

关键词:

钩藤碱 人星型胶质细胞 炎症 mTOR/STAT3信号通路

rhynchophylline human astrocyte inflammation mTOR/STAT3 signaling pathway

分类号:

R285 [医药卫生—中药学]

钩藤中钩藤碱对肿瘤坏死因子-α诱导人星型胶质细胞炎症模型作用机制研究.pdf