摘要
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.LPS0.5 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0.5 mg.L-1;3.LPS1.0 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.LPS0.5 mg.L-124 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.LPS1.0 mg.L-124 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 LPS0.5 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。
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（Pa0.05）.There were significantly positive correlations in the expressions between IL-1,IL-6,NF-κB and TPO and their mRNA（Pa0.01）.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.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2011年第10期784-786,共3页
Journal of Applied Clinical Pediatrics
关键词
小胶质细胞
促血小板生成素
脂多糖
炎症因子
microglial cell
thrombopoietin
lipopolysaccharide
inflammatory cytokines
