川陈皮素抑制 BV2 小胶质细胞炎症反应的机制.

Mechanism by which nobiletin inhibits inflammatory response of BV2 microglia.

BACKGROUND: Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia, excessive release of inflammatory factors and redox imbalance. However, the specific mechanism is not fully understood. OBJECTIVE: To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS: Passage 3 BV2 microglia were divided into three groups: control group was cultured for 24 hours (without any treatment). Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours. Lipopolysaccharide + nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours. After the processing, cell proliferation was detected by CCK-8 assay. The level of intracellular reactive oxygen species was detected by fluorescent probe. The mRNA expression levels of nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β were detected by qRT-PCR. The protein expression levels of nuclear factor κB p65, p-nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, the proliferation activity of lipopolysaccharide group was decreased (P < 0.001). Compared with the lipopolysaccharide group, the cell proliferation activity of lipopolysaccharide + nobiletin group was increased (P < 0.001). Compared with the control group, the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group (P < 0.001). Compared with the lipopolysaccharide group, the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide + nobiletin group (P < 0.01). Compared with the control group, the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group (P < 0.001, P < 0.01). Compared with the lipopolysaccharide group, mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide + nobiletin group (P < 0.01, P < 0.05). Compared with the control group, the protein expression levels of p-nuclear factor κB p65, tumor necrosis factor α, and interleukin1β in were increased the lipopolysaccharide group (P < 0.001). Compared with the lipopolysaccharide group, the expression of p-nuclear factor κB p65, tumor necrosis factor α, and interleukin-1β was decreased in the lipopolysaccharide + nobiletin group (P < 0.001). These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway. [ABSTRACT FROM AUTHOR]

背景: 有研究证实, 川陈皮素可改善脂多糖诱导的小胶质细胞异常激活、炎症因子的过度释放和氧化还原失衡, 但具体的作用机制尚不充分。 目的: 探讨川陈皮素抑制脂多糖诱导BV2小胶质细胞炎症反应的分子机制。方法: 将第 3 代BV2小胶质细胞分3组处理: 对照组常规培养24 h (不进行任何处理), 脂多糖组加入10 μg/mL脂多糖处理24 h, 脂多糖+川陈 皮素组加入20 μmol/L川陈皮素处理6 h后加入10 μg/mL脂多糖处理24 h。处理结束后, 采用CCK-8法检测细胞增殖, 活性氧荧光探针检测细 胞内活性氧水平, qRT-PCR法检测细胞内核因子κB p65、肿瘤坏死因子α、白细胞介素1β mRNA表达, Western Blot法检测细胞内核因子κB p65、p-核因子κB p65、肿瘤坏死因子α、白细胞介素1β蛋白表达。结果与结论: 与对照组比较, 脂多糖组细胞增殖活性降低 (P < 0.001); 与脂多糖组比较, 脂多糖+川陈皮素组细胞增殖活性升高 (P < 0.001); 与对照组比较, 脂多糖组细胞内活性氧水平升高 (P < 0.001); 与脂多糖组比较, 脂多糖+川陈皮素组细胞内活性氧水平降低 (P < 0.01); 与对照组比较, 脂多糖组细胞内肿瘤坏死因子α、白细胞介素1β mRNA表达升高 (P < 0.001, P < 0.01); 与脂多糖组比较, 脂多糖+川 陈皮素组细胞内肿瘤坏死因子α、白细胞介素1β mRNA表达降低 (P < 0.01, P < 0.05); 与对照组比较, 脂多糖组细胞内p-核因子κB p65、肿 瘤坏死因子α、白细胞介素1β蛋白表达升高 (P < 0.001); 与脂多糖组比较, 脂多糖+川陈皮素组细胞内p-核因子κB p65、肿瘤坏死因子α、白 细胞介素1β蛋白表达降低 (P < 0.001); 结果表明, 川陈皮素可能通过抑制核因子κB信号通路减轻脂多糖诱导的BV2小胶质细胞炎症反应。 [ABSTRACT FROM AUTHOR]

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