茯苓酸对TNF-α诱导SH-SY5Y细胞炎症氧化应激反应和凋亡的拮抗作用研究  

Effects of Pachymic Acid on Inflammation,Oxidative Stress Injury and Apoptosis of SH-SY5Y Cells Induced by TNF-α

作　　者：秦劭晨[1] 王爱梅[1] 李若瑜 QIN Shaochen;WANG Aimei;LI Ruoyu(The Affiliated Hospital of Shanxi College of Chinese Medicine,Taiyuan,Shanxi 030024)

机构地区：[1]山西中医学院附属医院

出　　处：《中国中医药科技》2019年第6期854-858,共5页Chinese Journal of Traditional Medical Science and Technology

基　　金：山西省卫生和计划生育委员会科研课题(201601119);山西中医药大学科技创新能力培育计划项目(2019PY-051)

摘　　要：目的:探讨茯苓酸对肿瘤坏死因子α(TNF-α)诱导SH-SY5Y细胞炎症、氧化应激损伤和凋亡的作用机制。方法:体外培养SH-SY5Y细胞,用TNF-α诱导SH-SY5Y造模,将细胞分为正常组、模型组、茯苓酸组、依达拉奉组。采用噻唑蓝比色(MTT)法检测细胞增殖能力,用比色法检测乳酸脱氢酶(LDH),超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;用酶联免疫吸附法(ELISA)检测白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的含量;用蛋白免疫印迹法(Western blot)检测B淋巴细胞瘤-2基因(BCL-2)、Bcl-2相关X蛋白(BAX)、磷酸化细胞外信号调节激酶(p-ERK)和核因子E2相关因子2(nuclear Nrf2)的蛋白表达。结果:在50 mg/L浓度时,TNF-α对SH-SY5Y细胞有明显的抑制作用(P<0.01)。与模型组比较,茯苓酸组和依达拉奉组SH-SY5Y细胞的LDH、MDA和IL-6、IL-1β的含量及BAX,p-ERK和nuclear Nrf2的表达显著减少(P<0.05或P<0.01),而SOD含量和BCL-2表达显著增加(P<0.05或P<0.01)。结论:茯苓有效成分茯苓酸具有抑制TNF-α诱导SH-SY5Y的炎症和氧化应激损伤作用,可能是通过下调ERK/Nrf2信号通路的Nrf2进入细胞核达到抑制细胞凋亡的作用机制。Objective:To explore the effects of Pachymic acid in Poriacocos on inflammation,oxidative stress injury and apoptosis of SH-SY5Y cells induced byTNF-α.Methods:SH-SY5Y cells were cultured in vitro and SH-SY5Y was induced with TNF-α.The cells were divided into normal group,model group,Pachymic acid group,and Edaravone group.The ability of cell proliferation was detected by MTT method and the content of lactate dehydrogenase(LDH),superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by colorimetric method.The levels of interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA),and the protein expressions of BCL-2,BAX,p-ERK and nuclear Nrf2 were detected by Western blot.Results:At the concentration of 50 mg/L,TNF-αhad a significant inhibitory effect on SH-SY5Y cells proliferation(P<0.01),compared with the model group,the contents of LDH,MDA,IL-6 and IL-1βand the expressions of BAX,p-ERK and nuclear Nrf2 were significantly decreased in the Pachymic acid group and Edaravone group(P<0.05 or P<0.01),while SOD activity and BCL-2 expression were significantly increased(P<0.05 or P<0.01).Conclusion:Pachymic acid has inhibitory effects on TNF-α-induced SH-SY5Y inflammation and oxidative stress injury,possibly through inhibition of apoptosis by Nrf2 down-regulation of the ERK/Nrf2 signaling pathway into the nucleus.

关 键 词：茯苓酸 肿瘤坏死因子 SH-SY5Y细胞 炎症 氧化应激 ERK/Nfr2通路 凋亡 体外实验 

分 类 号：R28[医药卫生—中药学]