醒脑益智组分中药对β-淀粉样蛋白_(1-40)诱导SH-SY5Y细胞损伤的保护作用
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摘要
目的:探讨醒脑益智组分中药对β-淀粉样蛋白(Aβ_(1-40))诱导SH-SY5Y细胞损伤的影响。方法:以SH-SY5Y细胞体外培养为研究对象,Aβ_(1-40)损伤细胞建立模型,石杉碱甲(6μg·mL~(-1))和不同浓度(70,35,17.5μg·mL~(-1))药物干预,四甲基偶氮唑盐(MTT)法检测细胞存活率;细胞膜损伤测定乳酸脱氢酶(LDH)漏出率;酶联免疫吸附法检测脑源性神经营养因子(BDNF)和肿瘤坏死因子(TNF-α)含量;Hochest 33258染色观察药物对损伤细胞的作用;AnnexinⅤ-FITC/PI双染流式细胞术检测细胞凋亡情况。结果:醒脑益智组分中药能明显改善Aβ_(1-40)(15μmol·L~(-1))诱导的神经细胞存活率,降低LDH漏出率,改善细胞内BDNF含量,降低TNF-α水平,并抑制细胞的凋亡。结论:醒脑益智组分中药可以改善Aβ_(1-40)诱导的SH-SY5Y细胞损伤,并抑制其凋亡,其机制可能与改善BDNF水平,调节免疫及相关凋亡基因的表达有关。
OBJECTIVE To study the effects of mental refreshing and improving components of TCM drugs on SH-SY5Y cell injury induced byβ-amyloid peptide 1-40.METHODS SH-SY5Y cells were cultured in vitro,with Aβ_(1-40) contained for preparing injury models and different concentrations of huperzine A(6μg·mL~(-1))and mental refreshing and improving components(70,35 and 17.5μg·mL~(-1))were used for intervention.Cell viability was measured by tetramethyl triazole salt(MTT)assay and cell membrane damage was detected by LDH leakage.The contents of the brain derived neurotrophic factors(BDNF)and tumor necrosis factor(TNF-α)were measured by ELISA.The injuried cells were observed by Hochest 33258 staining.Cell apoptotic rates were analyzed with AnnexinⅤ-FITC/PI flow cytometry.RESULTS Compared with model group,mental refreshing and improving components obviously improved the cell viability after injuried by Aβ_(1-40),decreased the LDH leakage,increased the content of BDNF,inhibited the increased TNF-αlevel and cell apoptotic rate induced by Aβ_(1-40).CONCLUSIONMental refreshing and improving components can allevaite damage and reduce apoptosis of SH-SY5Y cells induced by Aβ_(1-40),which may be related to BDNF adjustment,immune function regulation and apoptosis related protein expression.
OBJECTIVE To study the effects of mental refreshing and improving components of TCM drugs on SH-SY5Y cell injury induced byβ-amyloid peptide 1-40.METHODS SH-SY5Y cells were cultured in vitro,with Aβ_(1-40) contained for preparing injury models and different concentrations of huperzine A(6μg·mL~(-1))and mental refreshing and improving components(70,35 and 17.5μg·mL~(-1))were used for intervention.Cell viability was measured by tetramethyl triazole salt(MTT)assay and cell membrane damage was detected by LDH leakage.The contents of the brain derived neurotrophic factors(BDNF)and tumor necrosis factor(TNF-α)were measured by ELISA.The injuried cells were observed by Hochest 33258 staining.Cell apoptotic rates were analyzed with AnnexinⅤ-FITC/PI flow cytometry.RESULTS Compared with model group,mental refreshing and improving components obviously improved the cell viability after injuried by Aβ_(1-40),decreased the LDH leakage,increased the content of BDNF,inhibited the increased TNF-αlevel and cell apoptotic rate induced by Aβ_(1-40).CONCLUSIONMental refreshing and improving components can allevaite damage and reduce apoptosis of SH-SY5Y cells induced by Aβ_(1-40),which may be related to BDNF adjustment,immune function regulation and apoptosis related protein expression.
引文
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[3]李鹏,黄福开,杨春,等.中药对Aβ清除作用的研究进展[J].中国中药杂志,2013,38(23):4020-4023.
[4]胡海燕,金国琴,张学礼,等.中药对Aβ25-35损伤的SH-SY5Y细胞的保护作用[J].上海中医药大学学报,2013,27(5):77-81.
[5]张晓平,侯林,张龙霏,等.醒脑益智胶囊不同配比对阿尔茨海默病大鼠行为学的实验研究[J].中国实验方剂学杂志,2013,19(19):221-224.
[6]张晓平,侯林,聂克,等.醒脑益智胶囊对东莨菪碱致小鼠记忆获得障碍模型的影响[J].中国实验方剂学杂志,2013,19(23):228-230.
[7]黄炎,陈逸青,陈勤,等.远志皂苷对Aβ1-40诱导痴呆大鼠海马神经细胞线粒体的保护作用[J].中药药理与临床,2015,31(1):93-97.
[8]马涛,辛文锋,张文生,等.三七皂苷R1对Aβ1-42诱导的SHSY5Y细胞凋亡的保护作用[J].中国中药杂志,2015,40(2):303-307.
[9]苏萍,王蕾,杜仕静,等.三七皂苷对神经系统疾病药理作用机制研究进展[J].中国中药杂志,2014,39(23):4516-4521.
[10]张如意,张丽,艾厚喜,等.淫羊藿苷对线粒体损伤模型大鼠脑内β-淀粉样蛋白和神经营养因子的影响[J].中国中药杂志,2013,38(9):1285-1289.
[11]刘洪玲,郭素,王辉明,等.海康灵复方对Aβ1-40诱导SH-SY5Y细胞损伤的保护作用[J].中国医院药学杂志,2013,33(17):1405-1408.
[12]Swerdlow RH,Burns JM,Kham SM.The Alzheimer’s disease mitochondrial cascade hypothesis:progress and perspectives[J].Biochim Biophys Acta,2014,1842(8):1219.
[13]Hardy J,Selkoe DJ.The amyloid hypothesis of Alzheimer's disease progress and problems on the road to therapeutics[J].Science,2002,297(5580):353-356.
[14]Qin XY,Cao C,Cawley NX,et al.Decreased peripheral brain-derived neurotrophic factor levels in Alzheimer’s disease:a meta-analysis study[J].Molecular Psychiatry,2016,22(2):312.
[15]Zhao Y,Zhao B.Oxidative stress and the pathogenesis of Alzheimer's disease[J].Oxid Med Cell Longev,2013,2013(6):316-323.
[16]Packer N,Hoffman-Goetz L.Acute exercise increases hippocam-pal TNF-α,caspase-3 and Caspase-7 expression in healthy youngand older mice[J].J Sports Med Phys Fitness,2015,55:368-376.