糖皮质激素对巨噬细胞快速非基因组作用的研究

发布时间：2018-02-04 23:23

本文关键词： 糖皮质激素快速非基因组作用巨噬细胞吞噬功能超氧阴离子　出处：《青岛大学》2005年硕士论文　论文类型：学位论文

【摘要】：糖皮质激素(glucocorticoid,GC)的基因组作用机制虽然已得到公认,但是越来越多的证据表明,GC还存在着不同于经典基因组作用的非基因组作用。关于GC非基因组作用和机制的研究报道已有很多,但是大部分局限于神经和内分泌系统的调节作用。然而临床上GC最常应用于抗炎和抑制免疫治疗。迄今为止,关于GC的抗炎和抗免疫方面作用尚未见相关报道。巨噬细胞是体内参与炎症反应和免疫的重要细胞。本实验选择小鼠腹腔巨噬细胞,观察短时间(30min)内GC对巨噬细胞吞噬功能和产生超氧阴离子的影响,旨在探讨GC对巨噬细胞的非基因组作用的可能机制。本实验分离并纯化经巯基乙醇酸钠诱导的小鼠腹腔巨噬细胞,应用中性红(Neutral red,NR)吞噬分析法检测巨噬细胞的吞噬功能;细胞色素C(cytochrome C,Cyto C)还原法检测经PMA刺激后的巨噬细胞超氧阴离子的生成量。结果如下: (1) 10~(-4)mol/L和10~(-5)mol/L皮质酮(corticosterone,CORT)、甲基强的松龙(methylprednisolone,METH)和氢化考的松(hydrocortisone,HYDR)在30min内均能抑制巨噬细胞吞噬功能;任何浓度地塞米松(Dexamethone,DEXA)和10~(-6)mol/L CORT、METH、HYDY在30min内不能能抑制巨噬细胞吞噬功能。 (2)10~(-4)mol/L～10~(-10)mol/LCORT、METH、HYDR和DEXA在30min内均能抑制巨噬细胞产生超氧阴离子,且随着浓度的升高,其抑制作用增强。 (3)RU486(mifepristone)是GC经典受体(GCR)拮抗剂,可以与胞浆内的GC受体结合而阻断GC的基因组作用。GC基因组作用是通过最终效应蛋白发挥作用的,蛋白质合成抑制剂放线菌酮(actidion,ACTI)阻断GC的基因组作用。小牛血清白蛋白(Bovine Serum Albumin,BSA)是一种大分子物质,而实验证明BSA在30min内很少进入胞浆内,因此BSA偶联的皮质酮(BSA-CORT)在30min内不可能与胞浆受体结合而发挥基因组作用。在反应体系内分别加入RU486和ACTI,发现GC依旧能发挥上述快速抑制作用;BSA-CORT亦能在短时间(30min)内发挥上述快速抑制作用。
[Abstract]:Although the genomic mechanism of glucocorticoid glucocorticoid (GC) has been recognized, there is more and more evidence. There are also non-genomic interactions in GC which are different from those of classical genomes. There have been many reports on the non-genomic interaction and mechanism of GC. However, most of them are limited to the regulation of the nervous and endocrine system. However, GC is most commonly used in anti-inflammatory and immunosuppressive therapy. The anti-inflammatory and anti-immune effects of GC have not been reported yet. Macrophages are important cells involved in inflammatory response and immunity in vivo. In this experiment, mouse peritoneal macrophages were selected. The effects of GC on phagocytic function and superoxide anion production in macrophages were observed in a short period of 30 min. The aim of this study was to explore the possible mechanism of the non-genomic effect of GC on macrophages. The macrophages induced by sodium mercaptoglycolate were isolated and purified in this experiment. The phagocytic function of macrophages was detected by Neutral redder NR phagocytosis assay. Cytochrome cytochrome Cyto C reduction assay was used to detect the production of superoxide anion in macrophages stimulated by PMA. The results were as follows: (1) 10 ~ (-4) mol / L and 10 ~ (-5) mol / L ~ (L) corticosterone (Cort). Methyl prednisolone and hydrocortisone. HYDR could inhibit the phagocytosis of macrophages within 30 min. Any concentration of dexamethasone Dexamethoneum (DEXA) and 10- 6 mol / L CORTMETH. HYDY could not inhibit the phagocytosis of macrophages within 30 minutes. 10 / 10 / L / 10 / L / 10 / L / 10 / L / L / L / L / T / METH. Both HYDR and DEXA inhibited the production of superoxide anion in macrophages for 30 minutes, and the inhibitory effect increased with the increase of concentration. Ru486 mifepristone is a classical GC receptor GCR-antagonist. GC genomes can be blocked by binding to the GC receptor in the cytoplasm. GC genomes function through the final effector protein, an inhibitor of protein synthesis, actinomycin actidion. Bovine Serum albumin (BSA) is a macromolecule. The results showed that BSA rarely entered the cytoplasm within 30 minutes. Therefore, BSA coupled corticosterone (BSA-Cort) could not bind to cytoplasmic receptors and play a genomic role within 30 minutes. RU486 and ACTI were added into the reaction system, respectively. It was found that GC could still exert these rapid inhibitory effects. BSA-CORT can also play the role of rapid inhibition in a short time of 30 min.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33

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