本文關(guān)鍵詞： 糖皮質(zhì)激素 快速非基因組作用 巨噬細(xì)胞 吞噬功能 超氧陰離子　出處：《青島大學(xué)》2005年碩士論文　論文類型：學(xué)位論文

【摘要】：糖皮質(zhì)激素(glucocorticoid,GC)的基因組作用機(jī)制雖然已得到公認(rèn),但是越來越多的證據(jù)表明,GC還存在著不同于經(jīng)典基因組作用的非基因組作用。關(guān)于GC非基因組作用和機(jī)制的研究報道已有很多,但是大部分局限于神經(jīng)和內(nèi)分泌系統(tǒng)的調(diào)節(jié)作用。然而臨床上GC最常應(yīng)用于抗炎和抑制免疫治療。迄今為止,關(guān)于GC的抗炎和抗免疫方面作用尚未見相關(guān)報道。巨噬細(xì)胞是體內(nèi)參與炎癥反應(yīng)和免疫的重要細(xì)胞。本實(shí)驗(yàn)選擇小鼠腹腔巨噬細(xì)胞,觀察短時間(30min)內(nèi)GC對巨噬細(xì)胞吞噬功能和產(chǎn)生超氧陰離子的影響,旨在探討GC對巨噬細(xì)胞的非基因組作用的可能機(jī)制。 本實(shí)驗(yàn)分離并純化經(jīng)巰基乙醇酸鈉誘導(dǎo)的小鼠腹腔巨噬細(xì)胞,應(yīng)用中性紅(Neutral red,NR)吞噬分析法檢測巨噬細(xì)胞的吞噬功能;細(xì)胞色素C(cytochrome C,Cyto C)還原法檢測經(jīng)PMA刺激后的巨噬細(xì)胞超氧陰離子的生成量。結(jié)果如下: (1) 10~(-4)mol/L和10~(-5)mol/L皮質(zhì)酮(corticosterone,CORT)、甲基強(qiáng)的松龍(methylprednisolone,METH)和氫化考的松(hydrocortisone,HYDR)在30min內(nèi)均能抑制巨噬細(xì)胞吞噬功能;任何濃度地塞米松(Dexamethone,DEXA)和10~(-6)mol/L CORT、METH、HYDY在30min內(nèi)不能能抑制巨噬細(xì)胞吞噬功能。 (2)10~(-4)mol/L～10~(-10)mol/LCORT、METH、HYDR和DEXA在30min內(nèi)均能抑制巨噬細(xì)胞產(chǎn)生超氧陰離子,且隨著濃度的升高,其抑制作用增強(qiáng)。 (3)RU486(mifepristone)是GC經(jīng)典受體(GCR)拮抗劑,可以與胞漿內(nèi)的GC受體結(jié)合而阻斷GC的基因組作用。GC基因組作用是通過最終效應(yīng)蛋白發(fā)揮作用的,蛋白質(zhì)合成抑制劑放線菌酮(actidion,ACTI)阻斷GC的基因組作用。小牛血清白蛋白(Bovine Serum Albumin,BSA)是一種大分子物質(zhì),而實(shí)驗(yàn)證明BSA在30min內(nèi)很少進(jìn)入胞漿內(nèi),因此BSA偶聯(lián)的皮質(zhì)酮(BSA-CORT)在30min內(nèi)不可能與胞漿受體結(jié)合而發(fā)揮基因組作用。在反應(yīng)體系內(nèi)分別加入RU486和ACTI,發(fā)現(xiàn)GC依舊能發(fā)揮上述快速抑制作用;BSA-CORT亦能在短時間(30min)內(nèi)發(fā)揮上述快速抑制作用。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R33

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 孫照祝;哮喘及COPD病人氣道巨噬細(xì)胞吞噬功能及相關(guān)基因表達(dá)[D];山東大學(xué);2011年

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本文編號：1491446