蛋氨酸脑啡肽对树突状细胞表型及功能成熟的影响

发布时间：2018-04-28 04:27

本文选题：蛋氨酸脑啡肽 + 树突状细胞　；参考：《中国医科大学》2010年硕士论文

【摘要】： 前言 1975年Hughes等从猪脑提取液中分离出具有内源性吗啡样活性物质蛋氨酸脑啡肽(Methionine Enkephalin, MENK)。蛋氨酸脑啡肽的早期研究主要集中在作为镇痛剂和神经递质方面,但随着研究的深入,人们发现其受体在免疫系统以及某些肿瘤组织和正常组织细胞均有分布。MENK通过与免疫细胞上受体结合发挥免疫调节作用。树突状细胞(Dendritic cell, DC)是目前所知的机体内功能最强大的抗原提呈细胞,参与机体多种自身免疫性疾病、免疫缺陷病、肿瘤及一些炎症反应的病理过程,在机体的抗感染免疫及肿瘤免疫应答过程中起着极其重要的作用,是基础与临床研究的热点。未成熟DC在机体的外周组织中捕获和加工抗原,然后开始向淋巴器官转移,并渐进式成熟,与此同时DC表面重要的功能分子,如：MHC-Ⅱ、CD40和CD80(CD86)的表达增加。由于DC在免疫调节中的重要作用,使人们越来越多地关注DC的研究与应用,特别是在抗肿瘤方面的应用。因此,研究DC已经成为一个热点领域。鉴于MENK在免疫系统中的显著调节作用,所以研究MENK对DC的作用对于阐明MENK对免疫系统作用机理是十分必的,国内外未见报道。本研究就MENK对小鼠髓系DC2.4细胞株成熟过程中表型及功能的变化的影响作用进行探讨。方法采用本教研室保存并传代的小鼠髓系DC2.4细胞株进行实验,复苏培养细胞,应用扫描电镜技术、酸性磷酸酶活性检测、流式细胞仪检测技术、酶联免疫吸附试验等检测DC表型和功能的各种指标。结果 1、MENK处理的DC表面可见很长的突起；而未作处理的DC(即RPMI1640组)的突起很短甚至没有突起。 2、蛋氨酸脑啡肽组和LPS组的树突状细胞的酸性磷酸酶活性均低于RPMI1640组(P0.05)。 3、MENK和LPS类似,能够上调树突状细胞表面MHC-Ⅱ、CD86、CD40分子的表达。 4、ELISA结果显示DC在未做任何处理时(即RPMI 1640组)IL-12分泌的量少,MENK和LPS处理的DC能够分泌高水平的IL-12(P0.05)。结论 1、适宜浓度的蛋氨酸脑啡肽显著促进DC形态学的成熟。 2、适宜浓度的蛋氨酸脑啡肽作用使DC细胞内的酸性磷酸酶活性下降,表明DC吞噬抗原能力下降,递呈能力增加,趋向成熟。 3、适宜浓度的蛋氨酸脑啡肽显著增加DC的MHC-Ⅱ、CD40和CD86分子的表达,刺激DC发育成熟。 4、适宜浓度的蛋氨酸脑啡肽显著增加培养上清中IL-12的含量,说明MENK促进DC成熟的同时,上调DC的功能。
[Abstract]:Preface
An endogenous morphine like enkephalin (Methionine Enkephalin, MENK) was isolated from the extract of the pig brain in 1975. The early study of methionine enkephalin was mainly focused on as a analgesic and neurotransmitter, but as the study went deep, people found that their receptors were in the immune system and in some tumor groups. The tissues of the tissue and the normal tissue are distributed in.MENK by binding to the receptors on the immune cells. The Dendritic cell (DC) is the most powerful antigen presenting cell known in the body, and is involved in the pathological process of a variety of autoimmune diseases, pestilence defects, tumors and some inflammatory reactions. It plays an extremely important role in the process of anti infection immunity and tumor immune response in the body. It is a hot spot in basic and clinical research. Immature DC captures and processes antigens in the peripheral tissues of the body, and then begins to transfer to the lymphoid organs and gradually mature. At the same time, the important functional molecules on the surface of DC, such as MHC- II, CD40 and CD8 The expression of 0 (CD86) is increasing. Because of the important role of DC in immune regulation, people are paying more and more attention to the research and application of DC, especially in the application of anti-tumor. Therefore, the study of DC has become a hot field. In view of the significant adjustment of MENK in the immune system, the study of the effect of MENK on DC is to clarify MENK pairs. The mechanism of action of the immune system is very necessary and has not been reported at home and abroad. The effect of MENK on the changes of phenotype and function during the maturation of DC2.4 cell lines in mice was discussed.
Method
The mouse medullary DC2.4 cell line, which was preserved and passed by the Department of teaching and research, was used to carry out the experiment, to resuscitate the cultured cells, to use scanning electron microscopy (SEM), acid phosphatase activity detection, flow cytometry and enzyme linked immunosorbent assay to detect the various indexes of DC phenotypes and functions.
Result
1, MENK showed a long protuberance on the surface of DC, while the DC (RPMI1640 group) did not have a short protuberance.
2, the acid phosphatase activities of dendritic cells in methionine enkephalin group and LPS group were lower than those in group RPMI1640 (P0.05).
3, MENK and LPS can upregulate the expression of MHC- II, CD86 and CD40 molecules on the surface of dendritic cells.
4, ELISA results showed that DC did not produce any treatment (i.e. RPMI 1640 group) and IL-12 secreted less. DC treated by MENK and LPS could secrete high level IL-12 (P0.05).
conclusion
1, suitable concentration of methionine enkephalin significantly promoted the morphological maturation of DC.
2, the suitable concentration of methionine enkephalin could decrease the activity of acid phosphatase in DC cells, which showed that the ability of DC phagocytic antigen decreased and the ability of delivery increased and tended to mature.
3, the appropriate concentration of methionine enkephalin significantly increased the expression of MHC- II, CD40 and CD86 molecules in DC and stimulated the maturation of DC.
4, suitable concentration of methionine enkephalin significantly increased the content of IL-12 in the culture supernatant, indicating that MENK promoted DC maturation and increased DC function.

【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392.12

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