蒲葵子总黄酮对帕金森病细胞模型中人神经母细胞瘤细胞的保护作用  

Total flavonoids of Fructus Livistonae protect human neuroblastoma cells in a cell model of PD

作　　者：顾宏韬[1] 师会 王磊[1] 董雯[1] Gu Hongtao;Shi Hui;Wang Lei;Dong Wen(Department of Encephalopathy,Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine,Tianjin 300120,China)

机构地区：[1]天津市中医药研究院附属医院脑病科,300120

出　　处：《中华老年心脑血管病杂志》2021年第9期913-917,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的探讨蒲葵子总黄酮(TFFL)对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病细胞模型中人神经母细胞瘤细胞(SK-N-SH)损伤的影响。方法以正常培养的SK-N-SH作为正常组。SK-N-SH用2000μmol/L MPP+处理48 h建立帕金森病细胞模型,依据不同处理分为模型组、低浓度组(5μg/ml TFFL)、中浓度组(15μg/ml TFFL)、高浓度组(45μg/ml TFFL)、阴性组、干扰组(转染BLACAT1小分子干扰RNA)、对照组、转染组[转染微小RNA(miR)-29c-3p]、质粒组、载体组(转染pcDNA-BLACAT1)。试剂盒检测丙二醛(MDA)、乳酸脱氢酶(LDH)、还原性谷胱甘肽(GSH)水平;甲基噻唑基四唑检测细胞增殖;流式细胞术检测细胞凋亡率;qRT-PCR法检测BLACAT1、miR-29c-3p表达量。结果与正常组比较,模型组细胞存活率和GSH水平降低,miR-29c-3p表达降低(0.29±0.05 vs 1.00±0.03,P<0.05),MDA水平和LDH活性升高,细胞凋亡率升高,BLACAT1表达水平升高(4.01±0.42 vs 1.00±0.04,P<0.05)。低、中、高浓度组较模型组;干扰组较阴性组;转染组较对照组细胞存活率升高,GSH水平升高,MDA水平和LDH活性降低,细胞凋亡率降低(P<0.05,P<0.01)。BLACAT1可靶向调控miR-29c-3p;BLACAT1过表达逆转TFFL对MPP+诱导的细胞增殖、凋亡及氧化应激的作用。结论TFFL可通过调控BLACAT1/miR-29c-3p分子轴,抑制MPP+诱导的SK-N-SH氧化应激、凋亡及促进细胞增殖,进而保护神经细胞。Objective To study the effect of total flavonoids of Fructus Livistonae(TFFL)on SK-N-SH cell damage in a cell model of PD induced by 1-methyl-4-phenylpyridine ion(MPP+).Methods The normally cultured cells served as a normal group.A cell model of PD was estab-lished by treating the SK-N-SH cells in 2000μmol/L MPP+for 48 h.The cells were divided into model group,low concentration group,medium concentration group,high concentration group,negative group,intervention group,control group,transfection group,plasmid group,and vector group.The levels of MDA,LDH,and GSH were measured with an agent kit.The survival rate of cells was tested in MTT experiment.The apoptosis rate of cells was assayed by flow cytometry.The expressions of BLACAT1 and miR-29c-3p were detected by qRT-PCR.Results The survival rate of cells,level of GSH and expression level of miR-29c-3p(0.29±0.05 vs 1.00±0.03)were significantly lower while the level of MDA,activity of LDH,apoptosis rate of cells and expression level of BLACAT1(4.01±0.42 vs 1.00±0.04)were significantly higher in model group than in normal group(P<0.05).The survival rate of cells and level of GSH were significantly higher while the level of MDA and activity of LDH were significantly lower in low,medium and high concention groups than in model group,in intervention group than in negative group,and in trans-fection group than in control group(P<0.05,P<0.01).BLACAT1 could targetly regulate the miR-29c-3p.The overexpression of BLACAT1 could reverse the effect of TFFL on MPP+-in-duced proliferation,apoptosis and oxidative stress of cells.Conclusion TFFL can inhibit MPP+-induced oxidative stress and apoptosis of cells by regulating the molecular axis of BLACAT1/miR-29c-3p and promoting the proliferation of cells,thus playing a role in protecting nerve cells.

关 键 词：帕金森病 RNA 长链非编码 植物提取物 细胞凋亡 氧化性应激 

分 类 号：R285.5[医药卫生—中药学]