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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 investigate the expression of intracellular thrombopoietin (TPO) and inflammatory cytokines after lipopolysaccharide (LPS) intervention in microglia. Methods BV2 cells were divided into 6 groups. 1. Blank 12 h group: BV2 cells cultured for 12 h, without any intervention; 2. LPS0.5 mg.L-112 h group: cultured BV2 cells were added with pre-LPS solution co-culture 12 h, and its final concentration of 0.5 mg.L-1; 3. LPS1.0 mg.L-112 h group: cultured BV2 cells pre-mixed with LPS solution was added for 12 h, and The final concentration of 1.0 mg.L-1; 4. Blank 24 h group: BV2 cells cultured for 24 h without any intervention; 5. LPS0.5 mg.L-124 h group: cultured BV2 The cells were pre-incubated with LPS solution for 24 h and the final concentration was 0.5 mg.L-1. 6. LPS1.0 mg.L-124 h group: cultured BV2 cells were added prior LPS solution with a good co-culture 24 h, and the final concentration of 1.0 mg.L-1. The expression of inflammatory cytokines (IL-1, IL-6, NF-κB) and TPO in BV2 cells were detected by ELISA. The levels of IL-1 mRNA and IL-6 mRNA and NF-κB mRNA in BV2 cells were detected by real-time fluorescence quantitative PCR TPO mRNA expression level. Results The expression levels of IL-1, IL-6, NF-κB and TPO at 12 h and 24 h after LPS 0.5 mg · L-1 and 1.0 mg · L-1 intervention were higher than those in blank group High, and 12 h group was higher than 24 h group, but the difference was not statistically significant (Pa> 0.05). The expression of TPO was positively correlated with the expression of IL-1, IL-6, NF-κB and its mRNA (Pa <0.01). CONCLUSIONS: The expression of IL-1, IL-6, NF-κB and TPO in LPS-treated microglial cells is increased, which is involved in many regulatory mechanisms such as inflammation, apoptosis and neuronal protection.