谷氧还蛋白在牙龈卟啉单胞菌脂多糖诱导脐静脉内皮细胞氧化应激中的作用

发布时间：2018-03-20 20:58

本文选题：牙龈卟啉单胞菌　切入点：脂多糖　出处：《华西口腔医学杂志》2015年06期 　论文类型：期刊论文

【摘要】：目的从基因和蛋白水平研究谷氧还蛋白(Grx)在牙龈卟啉单胞菌脂多糖(LPS)诱导脐静脉内皮细胞(EA-hy926细胞)时的表达变化及其对Akt通路的调控作用。方法采用牙龈卟啉单胞菌的LPS(1 000 ng·m L-1)对EA-hy926细胞进行不同时间段(4、12、18、24 h)的刺激诱导,采用实时荧光定量逆转录聚合酶链反应检测细胞grx1基因的表达变化;然后加入Grx特异性抑制剂氯化亚硝脲(BCNU),使用Western blot法检测对照组、LPS组(1 000 ng·m L~(-1)LPS刺激12 h)和BCNU组(25μmol·m L-1BCNU预处理30 min+1 000 ng·m L~(-1)LPS刺激12 h)的Grx、Akt、磷酸化Akt蛋白的表达情况。结果 LPS诱导下,EA-hy926细胞的grx1基因表达量在各个时间段均上调,12 h时grx1表达量最高。LPS诱导12 h时,LPS组的Grx蛋白表达量较对照组明显升高(P0.05),而BCNU可有效抑制Grx蛋白的表达(P0.05);Akt蛋白表达量在各组间无明显差异(P0.05),而LPS组的磷酸化Akt活性升高,显著高于对照组和BCNU组(P0.05),与Grx蛋白的表达趋势一致。结论 LPS可以从基因和蛋白水平诱导Grx的表达;Grx是Akt的潜在调节因子,可能对LPS刺激下Akt的调控具有重要意义。
[Abstract]:Objective to investigate the expression of glutathione (Grx) in cultured umbilical vein endothelial cells (EA-hy926) induced by lipopolysaccharide (LPS) from porphyromonas gingivalis at the gene and protein levels. Methods the regulation of Akt pathway was studied by using gingival porphyrin monomonas. EA-hy926 cells were stimulated by LPS(1 (#number0# ng 路m -1) for 1824 h. The expression of grx1 gene was detected by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction. Western blot assay was used to detect the expression of phosphorylated Akt protein in control group (1 000 ng 路mL ~ (-1)) and BCNU group (n = 25 渭 mol 路ml ~ (-1) BCNU for 12 h) treated with 30 min 1 000 ng 路mL ~ (-1) BCNU for 12 h. The expression of grx1 gene in fruit LPS induced HY926 cells was up-regulated at each time point at 12 h. The expression of Grx protein in lipopolysaccharide group was significantly higher than that in control group at 12 h. BCNU could effectively inhibit the surface of Grx protein. There was no significant difference in the expression of P0.05Akt protein among the groups, but the phosphorylated Akt activity in LPS group was higher than that in the control group. Conclusion LPS can induce the expression of Grx from gene and protein levels, which may be a potential regulatory factor for Akt, and may play an important role in the regulation of Akt stimulated by LPS.
【作者单位】：口腔疾病研究国家重点实验室华西口腔医院牙周病科(四川大学);
【基金】：国家自然科学基金资助项目(81371150) 四川省科技计划基金资助项目(2011JY0128)
【分类号】：R780.2

【共引文献】