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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。
AIM: To investigate the effects of berberine on the expression of tumor necrosis factor α (TNF-α) and type Ⅰ receptor (TNFR1) in the inflammatory response induced by β-amyloid peptide (Aβ) in SH-SY5Y cell line. Methods: SH-SY5Y cells were treated with 5μmol.L-1Aβ25-35 for 24 h, and Alzheimer’s disease (AD) cells were replicated. Pretreatment with berberine for 2 h was performed. The experiment was divided into normal control group, AD model group, indomethacin group, berberine low and high dose group. The activity of LDH in culture solution was detected by spectrophotometry. The level of TNF-α was measured by ELISA. The expression of TNFR1 gene was detected by RT-PCR. The expression of TNFR1 protein was detected by Western blot. Results: Compared with the control group, the levels of LDH and TNF-α in the culture medium of AD model group were significantly increased, and the expression of TNFR1 gene and protein was significantly increased. Berberine intervention in the cell supernatant LDH activity and TNF-α levels decreased. Berberine can down-regulate the expression of TNFR1 gene and protein, and the effect of 1 × 10-5mol.L-1 berberine on TNFR1 is more significant. CONCLUSION: Berberine can protect Aβ-induced inflammatory injury in SH-SY5Y cells in a dose-dependent manner by inhibiting the expression of TNF-α and its type Ⅰ receptor.