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  本文選題：蛋氨酸腦啡肽 + 樹突狀細胞��； 參考：《中國醫(yī)科大學》2010年碩士論文

【摘要】： 前言 1975年Hughes等從豬腦提取液中分離出具有內源性嗎啡樣活性物質蛋氨酸腦啡肽(Methionine Enkephalin, MENK)。蛋氨酸腦啡肽的早期研究主要集中在作為鎮(zhèn)痛劑和神經遞質方面,但隨著研究的深入,人們發(fā)現其受體在免疫系統以及某些腫瘤組織和正常組織細胞均有分布。MENK通過與免疫細胞上受體結合發(fā)揮免疫調節(jié)作用。樹突狀細胞(Dendritic cell, DC)是目前所知的機體內功能最強大的抗原提呈細胞,參與機體多種自身免疫性疾病、免疫缺陷病、腫瘤及一些炎癥反應的病理過程,在機體的抗感染免疫及腫瘤免疫應答過程中起著極其重要的作用,是基礎與臨床研究的熱點。未成熟DC在機體的外周組織中捕獲和加工抗原,然后開始向淋巴器官轉移,并漸進式成熟,與此同時DC表面重要的功能分子,如：MHC-Ⅱ、CD40和CD80(CD86)的表達增加。由于DC在免疫調節(jié)中的重要作用,使人們越來越多地關注DC的研究與應用,特別是在抗腫瘤方面的應用。因此,研究DC已經成為一個熱點領域。鑒于MENK在免疫系統中的顯著調節(jié)作用,所以研究MENK對DC的作用對于闡明MENK對免疫系統作用機理是十分必的,國內外未見報道。本研究就MENK對小鼠髓系DC2.4細胞株成熟過程中表型及功能的變化的影響作用進行探討。 方法 采用本教研室保存并傳代的小鼠髓系DC2.4細胞株進行實驗,復蘇培養(yǎng)細胞,應用掃描電鏡技術、酸性磷酸酶活性檢測、流式細胞儀檢測技術、酶聯免疫吸附試驗等檢測DC表型和功能的各種指標。 結果 1、MENK處理的DC表面可見很長的突起；而未作處理的DC(即RPMI1640組)的突起很短甚至沒有突起。 2、蛋氨酸腦啡肽組和LPS組的樹突狀細胞的酸性磷酸酶活性均低于RPMI1640組(P0.05)。 3、MENK和LPS類似,能夠上調樹突狀細胞表面MHC-Ⅱ、CD86、CD40分子的表達。 4、ELISA結果顯示DC在未做任何處理時(即RPMI 1640組)IL-12分泌的量少,MENK和LPS處理的DC能夠分泌高水平的IL-12(P0.05)。 結論 1、適宜濃度的蛋氨酸腦啡肽顯著促進DC形態(tài)學的成熟。 2、適宜濃度的蛋氨酸腦啡肽作用使DC細胞內的酸性磷酸酶活性下降,表明DC吞噬抗原能力下降,遞呈能力增加,趨向成熟。 3、適宜濃度的蛋氨酸腦啡肽顯著增加DC的MHC-Ⅱ、CD40和CD86分子的表達,刺激DC發(fā)育成熟。 4、適宜濃度的蛋氨酸腦啡肽顯著增加培養(yǎng)上清中IL-12的含量,說明MENK促進DC成熟的同時,上調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.

【學位授予單位】：中國醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R392.12

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