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目的:观察锌离子螯合剂TPEN对脂多糖(LPS)刺激下小鼠BV2小胶质细胞核转录因子NF-кB P65亚基磷酸化及诱导型一氧化氮合酶(iNOS)mRNA表达的影响。方法:体外培养BV2细胞,用不同药物进行孵育,分组如下:正常对照组,单独LPS组,TPEN与LPS共处理组,NF-кB抑制剂BAY11-7082和LPS共处理组。采用实时荧光定量PCR法检测iNOS mRNA表达,Western Blot法检测磷酸化NF-кB P65(p-P65)蛋白表达。结果:TPEN浓度依赖性地减少LPS诱导的p-P65蛋白和iNOS mRNA表达,BAY11-7082浓度依赖性抑制LPS诱导的iNOSmRNA表达的上调。结论:TPEN能够减少LPS诱导的BV2细胞iNOS mRNA表达,该作用可能与其抑制NF-кB信号通路的激活有关。
OBJECTIVE: To observe the effect of zinc ion chelator TPEN on the phosphorylation of NF-кB P65 and the expression of inducible nitric oxide synthase (iNOS) mRNA in BV2 microglia of mice induced by lipopolysaccharide (LPS). Methods: BV2 cells were cultured in vitro and incubated with different drugs. The groups were as follows: normal control group, LPS alone group, TPEN and LPS co-treatment group, NF-κB inhibitor BAY11-7082 and LPS co-treatment group. The expression of iNOS mRNA was detected by real-time fluorescence quantitative PCR. The protein expression of phosphorylated NF-кB P65 (p-P65) was detected by Western Blot. Results: TPEN concentration-dependently reduced the LPS-induced expression of p-P65 protein and iNOS mRNA. BAY11-7082 inhibited LPS-induced iNOS mRNA expression in a concentration-dependent manner. Conclusion: TPEN can reduce the LPS-induced iNOS mRNA expression in BV2 cells, which may be related to the inhibition of the activation of NF-κB signaling pathway.