中国中药杂志

2013, v.38(09) 1327-1330

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小檗碱对Aβ_(25-35)致SH-SY5Y细胞株炎症反应中TNF-α及Ⅰ型受体表达的干预作用
Intervention effect of berberine on expressions of TNF-α and receptor type I in Aβ_(25-35)-induced inflammatory reaction in SH-SY5Y cell lines

许旌;张洪;杨帆;俞金鑫;
XU Jing,ZHANG Hong,YANG Fan,YU Jin-xin(Department of Geriatrics,Drum Tower Hospital Affiliated to Medical School of Nanjing University,Najing 210008,China)

摘要(Abstract):

目的:研究小檗碱对β淀粉样肽(Aβ)沉积导致SH-SY5Y细胞株炎症反应中肿瘤坏死因子α(TNF-α)及Ⅰ型受体(TNFR1)表达的影响。方法:5μmol.L-1Aβ25-35作用于SH-SY5Y细胞24 h,复制阿尔茨海默病(AD)细胞模型,造模前给予小檗碱预处理2 h。实验分为正常对照组、AD模型组、吲哚美辛组、小檗碱低、高剂量组。通过分光光度法检测培养液中LDH的活力,ELISA测定TNF-α的水平,RT-PCR检测TNFR1基因的表达,Western blot测定TNFR1蛋白的表达。结果:与对照组比较,AD细胞模型组培养液中LDH及TNF-α水平明显升高,TNFR1基因和蛋白的表达显著增加。小檗碱干预后细胞上清液中LDH活力及TNF-α水平下降。小檗碱干预可下调TNFR1基因和蛋白的表达,其中1×10-5mol.L-1小檗碱对TNFR1的调节作用更加显著。结论:小檗碱对Aβ导致的SH-SY5Y细胞炎性损伤有保护作用,其机制可能与其抑制炎症因子TNF-α及其Ⅰ型受体的表达有关,其中对TNFR1的调节呈剂量依赖性。
Objective: To investigate the effect of berberine on expressions of tumor necrosis factor α(TNF-α) and receptor type I(TNFR1) in Aβ25-35-induced inflammatory reaction in SH-SY5Y cell lines.Method: The 5 μmol·L-1Aβ25-35 was used to treat SH-SY5Y cells for 24 hours,in order to establish the Alzheimer′s disease(AD) model.Before modeling,berberine was given for pre-treatment for 2 hours.The experiment included the normal control group,the AD model group,and indometacin low dose and high dose groups.Spectrophotometry was adopted to detect the activity of LDH.Meanwhile,the level of TNF-α was determined by ELISA,and the expression of TNFR1 genes was detected by RT-PCR.Result: Compared with the normal control group,the AD cell model group showed significant increase in LDH,TNF-α,and TNFR1 gene and protein expressions in the culture media.After intervention with berberine,the activity of LDH and TNF-α reduced in cell supernatant.The intervention with berberine could down-regulate TNFR1 gene and protein expressions,particularly 1,10×10-6 mol·L-1 berberine showed a more notable effect in regulating TNFR1.Conclusion: Berberine has the protective effect in Aβ-induced inflammatory injury in SH-SY5Y cells.Its mechanism may be related to the expression of its anti inflammatory factor TNF-α and its type I receptor TNFR1.Specifically,its regulation to TNFR1 shows dose dependence.

关键词(KeyWords): 阿尔茨海默病;β淀粉样肽;炎症反应;小檗碱;肿瘤坏死因子α;肿瘤坏死因子Ⅰ型受体
Alzheimer′s disease;Amyloid-β peptide;inflammatory reaction;berberine;tumor necrosis factor α;tumor necrosis factor type I receptor

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基金项目(Foundation): 南京市青年科技人才启动项目(QYK09170)

作者(Author): 许旌;张洪;杨帆;俞金鑫;
XU Jing,ZHANG Hong,YANG Fan,YU Jin-xin(Department of Geriatrics,Drum Tower Hospital Affiliated to Medical School of Nanjing University,Najing 210008,China)

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