糖皮质激素诱导的亮氨酸拉链蛋白（GILZ）与炎症反应关系的初步研究

发布时间：2019-03-12 12:36

【摘要】： 目的:初步探讨糖皮质激素诱导的亮氨酸拉链蛋白(GILZ)在炎症反应过程中的作用机理。方法:用无血清培养基分两组培养人单核细胞系THP-1细胞,其中一组加入地塞米松刺激,12小时后提取两组细胞总RNA和总蛋白,运用半定量逆转录-聚合酶链反应(RT-PCR)方法在转录水平检测GILZ基因;运用蛋白质印迹(Western blotting)方法在翻译水平检测转录因子NF- B与AP-1的表达。收集10例临床创伤患者伤后24小时内的外周血,提取白细胞后分两组给予无血清培养基,并在其中一组加入地塞米松刺激,培养12小时后提取白细胞总RNA和总蛋白,运用半定量RT-PCR方法在转录水平检测GILZ基因;运用Western blotting方法在翻译水平检测NF- B与AP-1的表达。结果:在地塞米松刺激下,刺激组THP-1细胞和创伤患者外周血白细胞的GILZ mRNA表达水平比对照组升高(P0.01);刺激组的NF- B与AP-1蛋白表达水平低于对照组(P0.01),差异具有极显著性。结论:糖皮质激素可上调GILZ基因的表达;GILZ的高表达可以抑制转录因子NF- B与AP-1的活性,具有抗炎症反应作用。
[Abstract]:Aim: to investigate the mechanism of glucocorticoid-induced leucine zipper (GILZ) in inflammatory reaction. Methods: two groups of human monocytes THP-1 cells were cultured in serum-free medium. One group was stimulated with dexamethasone and the total RNA and total protein were extracted 12 hours later. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect GILZ gene at transcription level. Western blot (Western blotting) was used to detect the expression of transcription factor NF- B and AP-1 at translation level. The peripheral blood of 10 patients with clinical trauma was collected within 24 hours after injury. After extracting white blood cells, they were divided into two groups and given serum-free medium, and one group was stimulated by dexamethasone. After 12 hours of culture, total RNA and total protein of white blood cells were extracted. Semi-quantitative RT-PCR was used to detect GILZ gene at transcription level. Western blotting method was used to detect the expression of NF- B and AP-1 at the translation level. Results: under the stimulation of dexamethasone, the expression of GILZ mRNA in THP-1 cells and peripheral blood leukocytes in the stimulation group was higher than that in the control group (P0.01). The expression level of NF- B and AP-1 protein in stimulation group was significantly lower than that in control group (P0.01). Conclusion: glucocorticoid can up-regulate the expression of GILZ gene, and the high expression of GILZ can inhibit the activity of transcription factor NF- B and AP-1, and has anti-inflammatory effect.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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