脂多糖干预小胶质细胞后细胞内促血小板生成素与炎症因子的表达

目的 探讨脂多糖(LPS)干预小胶质细胞后细胞内促血小板生成素(TPO)及炎症因子的表达.方法将BV2细胞分为6组.1.空白12 h组:BV2细胞正常培养12 h,不添加任何干预因素;2.LPS 0.5 mg·L-1 12 h组:在培养好的BV2细胞内添加预先配好的LPS溶液共同培养12 h,并使其终质量浓度为0.5 mg·L-1;3.LPS 1.0 mg·L-112 h组:在培养好的BV2细胞内添加预先配好的LPS溶液共同培养12 h,并使其终质量浓度为1.0 mg·L-1;4.空白 24 h组:BV2细胞正常培养24 h,不添加任何干预因素;5.LPS 0.5 mg·L-1 24 h组:在培养好的BV2细胞内添加预先配好的LPS溶液共同培养24 h,并使其终质量浓度为0.5 mg·L-1;6.LPS 1.0 mg·L-1 24 h组:在培养好的BV2细胞内添加预先配好的LPS溶液共同培养24 h,并使其终质量浓度为1.0 mg·L-1.采用ELISA法检测BV2细胞内炎症因子(IL-1、IL-6、NF-κB)和TPO的表达;实时荧光定量PCR法检测细胞内IL-1 mRNA、IL-6 mRNA、NF-κB mRNA及TPO mRNA表达水平.结果 LPS 0.5 mg·L-1和1.0 mg·L-1干预BV2细胞后12 h和24 h,IL-1、IL-6、NF-κB和TPO表达水平及其mRNA水平均较空白组升高,且12 h组高于24 h组,但其差异均无统计学意义(Pa>0.05).TPO表达水平与IL-1、IL-6、NF-κB及其mRNA水平的表达均呈显著正相关(Pa<0.01).结论 LPS干预小胶质细胞后IL-1、IL-6、NF-κB和TPO表达升高,参与炎症、凋亡和神经元保护等多种调控机制.

Expressions of Thrombopoietin and Inflammatory Cytokines in Microglial Cells Intervened with Lipopolysaccharide

Objective To study the expressions of thrombopoietin(TPO) and inflammatory cytokines in microglia cells intervened with lipopolysaccharide(LPS).Methods BV2 cells were randomly divided into 6 groups:1.with no additions for 12 h group:BV2 cells were cultured for 12 h without adding any intervention factors.2.LPS 0.5 mg·L-1 for 12 h group:pre-incubated LPS 0.5 mg·L-1 solution was added to culture BV2 cells with 12 hours′ cocultivation,resulting in a final concentration of 0.5 mg·L-1.3.LPS 1.0 mg·L-1 for 12 h group:pre-incubated LPS 1.0 mg·L-1 solutions was added to culture BV2 cells for 12 hours′ cocultivation,resulting in a final concentration of 1.0 mg·L-1.4.with no additions for 24 h group:BV2 cells were cultured for 24 h,without adding any intervention factors.5.LPS 0.5 mg·L-1 for 24 h group:pre-incubated LPS 0.5 mg·L-1 solutions was added to culture BV2 cells for 24 hours′ cocultivation,resulting in a final concentration of 0.5 mg·L-1.6.LPS 1.0 mg·L-1 for 24 h group:pre-incubated LPS 0.5 mg·L-1 solutions was added to culture BV2 cells with 24 hours′ cocultivation,resulting in a final concentration of 1.0 mg·L-1.The expressions of intracellular inflammatory cytokines such as IL-1,IL-6,nuclear factor-κB(NF-κB)] and TPO were detected by ELISA;Real-time quantitative PCR was used to check the expressions of IL-1 mRNA,IL-6 mRNA,NF-κB and TPO mRNA levels.Results The expressions of IL-1,IL-6,NF-κB and TPO and their mRNA levels in experimental groups(12 h,24 h) with the intervention of LPS(0.5 mg·L-1,1.0 mg·L-1) were higher significantly than those in the control groups,but the differences were not significant(Pa>0.05).There were significantly positive correlations in the expressions between IL-1,IL-6,NF-κB and TPO and their mRNA(Pa<0.01).Conclusions LPS can significantly increase the expressions of IL-1,IL-6,NF-κB and TPO in BV2 cells,which may participate in the inflammation,apoptosis and neuronal protection in central nervous system.