本文選題：骨髓增生異常綜合征 + 髓樣抑制細胞��； 參考：《河北醫(yī)科大學》2017年碩士論文

【摘要】：目的:骨髓增生異常綜合征(Myelodysplastic syndrome,MDS)是起源于造血干細胞的克隆性疾病,以一系或多系外周血細胞減少、骨髓無效和/或病態(tài)造血,并易向急性白血病(Acute leucemia,AL)轉(zhuǎn)化為特點。MDS的骨髓微環(huán)境是MDS形成和發(fā)展的關(guān)鍵機制,腫瘤細胞通過利用和改變微環(huán)境來逃避免疫殺傷而優(yōu)勢于正常細胞生長。髓樣抑制細胞(Myeloid-derived suppressor cells,MDSCs)是一群具有相似生物學功能的異質(zhì)性細胞,包括骨髓祖細胞、未成熟髓系細胞(Immature myeloid cells,IMCs)、未成熟的巨噬細胞和未成熟的樹突狀細胞。MDSCs可通過調(diào)節(jié)固有免疫及適應性免疫應答促進腫瘤發(fā)生免疫逃逸,如抑制CD4+T細胞、CD8+T細胞和自然殺傷細胞(Natural kill cells,NK)等的活化及免疫應答。腫瘤微環(huán)境和MDSCs誘導的免疫耐受相互作用來促進MDS的發(fā)展。因MDS多見于老年人,所以不能承受強烈化療及行異基因造血干細胞移植(Allogeneic hematopoietic stem cell transplantation,Allo-HSCT),尤其對于高危的MDS患者,極易向白血病轉(zhuǎn)化,故預后極差。對于不適用行Allo-HSCT的患者,仍以藥物治療為主。MDS的藥物治療包括去甲基化藥物、組蛋白去乙�；敢种苿�、鐵螯合劑及抗血管增殖藥物等。盡管這些藥物已經(jīng)應用于臨床治療MDS,但這些藥物都沒有獲得持久療效。為中高�；颊邔ふ业投�、有效、經(jīng)濟的藥物治療是十分需要解決的臨床問題。青蒿琥酯(Artesunate,ART)是一種從中藥青蒿中提取出來的青蒿素的半合成衍生物,是治療惡性瘧疾最有效的低毒的藥物之一,研究發(fā)現(xiàn)其除具有抗瘧作用外,在體外研究中還發(fā)現(xiàn)其具有抗腫瘤作用,ART可誘導MDS細胞系SKM-1細胞凋亡,并且ART與傳統(tǒng)化療藥物的殺傷機制不同,可能與其他MDS治療藥物存在協(xié)同作用。并且ART在治療腫瘤方面具有顯著的優(yōu)勢,如價格低廉,高效低毒、腫瘤靶向性等。本實驗通過研究MDS患者外周血MDSCs、CD34+細胞、NK細胞數(shù)目以及與MDS疾病惡性程度之間的關(guān)系。通過體外細胞培養(yǎng)實驗,檢測ART對MDS來源的MDSCs、NK細胞、CD34+細胞的各細胞群的變化,對進一步研究MDS的發(fā)病機制,ART抗MDS機制有十分重要的臨床意義,對尋找新的藥物治療MDS具有重要意義。方法:選擇2016年4月至2016年12月于河北醫(yī)科大學第二醫(yī)院血液內(nèi)科初治的MDS患者20例以及健康體檢者10例為實驗對象。同時將MDS患者行IPSS評分對疾病惡性程度進行分層分析。取實驗對象外周血各2ml,采用流式細胞術(shù)(Flow cytometry,FCM)檢測其外周血中MDSCs(CD33+CD11b+CD14-HLA-DR-)、NK細胞(CD3-CD56+)和CD34+細胞比例。抽取3例MDS患者外周血各20ml,提取外周血單個核細胞(Peripheral blood mononuclear cells,PBMCs),并接種于96孔板,用不同濃度的ART(終濃度分別為1、2.5、5、10umol/L),處理相應各孔48小時,用FCM檢測MDSCs、NK細胞、CD34+各群細胞的增殖和凋亡變化。結(jié)果:1健康者外周血MDSCs比例(0.65%±0.28%)明顯低于MDS患者(2.39%±0.74%)(P0.05),健康者外周血MDSCs比例(0.65%±0.28%)低于低危及中危-ⅠMDS患者(1.46%±0.17%)(P0.05);低危及中危-ⅠMDS患者外周血MDSCs比例(1.46%±0.17%)低于中危-Ⅱ及高危MDS患者(2.69%±0.56%)(P0.05)。2健康者外周血NK細胞占淋巴細胞百分比(12.88%±4.51%)明顯高于MDS患者(9.08%±3.59%)(P0.05);健康者外周血NK細胞占淋巴百分比(12.88%±4.51%)與低危及中危-ⅠMDS患者(13.71%±2.46%)相比,差異無統(tǒng)計學意義(P0.05);低危及中危-ⅠMDS患者外周血NK細胞占淋巴細胞百分比(13.71%±2.46%)高于中危-Ⅱ及高危MDS患者(7.53%±2.36%)(P0.05)。3健康者外周血CD34+細胞百分數(shù)(0.03%±0.01%)明顯低于MDS患者(3.75%±3.12%)(P0.05);健康者外周血CD34+細胞百分數(shù)(0.03%±0.01%)低于低危及中危-ⅠMDS患者(0.34%±0.36%)(P0.05);低危及中危-ⅠMDS患者(0.34%±0.36%)低于中危-Ⅱ及高危MDS患者(4.88%±2.77%)(P0.05)。4不同濃度的ART對MDS外周血來源的MDSCs、NK細胞、CD34+細胞的增殖和凋亡均未見明顯差異(P0.05)。結(jié)論:1 MDS患者外周血循環(huán)中MDSCs和CD34+細胞比例均是升高的,且隨著疾病危險程度增加而增高;其NK細胞比例是降低的,并隨著疾病危險程度增加而降低。2 ART對MDS外周血來源的MDSCs、NK細胞、CD34+細胞的增殖和凋亡暫未見明顯影響。
[Abstract]:Objective: Myelodysplastic syndrome (MDS) is a clonogenic disease originating from hematopoietic stem cells. It is a key mechanism for the formation and development of MDS to reduce the blood cells of one or more peripheral peripheral blood, bone marrow ineffective and / or morbid hematopoiesis, and the transformation of Acute leucemia (AL) to the characteristic.MDS in the bone marrow microenvironment. Myeloid-derived suppressor cells (MDSCs) is a group of heterogeneous cells with similar biological functions, including bone marrow progenitor cells, immature myeloid cells (Immature myeloid cells, IMCs), immature macrophages, by using and changing microenvironment to escape immune killing. Cells and immature dendritic cells.MDSCs can promote immune escape by regulating inherent immunity and adaptive immune response, such as inhibition of CD4+T cells, CD8+T cells and natural killer cells (Natural kill cells, NK) and other activation and immune responses. The interaction of tumor microenvironment and MDSCs induced immune tolerance promotes MDS Development. Because MDS is more common in the elderly, it can not bear strong chemotherapy and transgene hematopoietic stem cell transplantation (Allogeneic hematopoietic stem cell transplantation, Allo-HSCT), especially for high-risk MDS patients, it is very easy to convert to leukemia, so the prognosis is extremely poor. For patients who do not apply Allo-HSCT, they are still treated with drug therapy as the main.MDS. Drug treatment includes demethylation drugs, histone deacetylase inhibitors, iron chelating agents and antiproliferative drugs. Although these drugs have been used in clinical treatment of MDS, these drugs have not achieved a lasting effect. It is a very important clinical question to find low toxicity, effective and economic treatment for middle risk patients. Artesunate (ART) is a semi synthetic derivative of artemisinin extracted from the Chinese herb Artemisia Artemisia. It is one of the most effective and low toxic drugs for the treatment of malarial malaria. It is found that it has anti swelling effect in vitro, and ART can induce apoptosis of SKM-1 cells in MDS cell line in vitro. And ART is different from the traditional chemotherapeutic drugs, and may have synergistic effects with other MDS drugs. And ART has significant advantages in the treatment of tumor, such as low price, high efficiency and low toxicity, and tumor targeting. In this study, the number of MDSCs, CD34+ cells, the number of NK cells and the malignancy of MDS disease in the peripheral blood of MDS patients were studied. Through in vitro cell culture experiment, it is important to detect the changes in the cell groups of MDSCs, NK and CD34+ cells derived from MDS by ART. It is of great significance to further study the pathogenesis of MDS and the mechanism of ART anti MDS, and is of great significance for finding new drugs to treat MDS. Method: choose from April 2016 to December 2016. In the second hospital of the second hospital of Northern Medical University, 20 cases of MDS and 10 healthy subjects were treated as subjects. At the same time, the malignant degree of the disease was stratified by IPSS score of the MDS patients. The peripheral blood of the experimental subjects was taken 2ml, and MDSCs (CD33+CD11b+CD14-HLA-DR-) in peripheral blood was detected by flow cytometry (FCM), and the flow cytometry (FCM) was used to detect MDSCs (CD33+CD11b+CD14-HLA-DR-) in peripheral blood. The proportion of NK cells (CD3-CD56+) and CD34+ cells. The peripheral blood of 3 MDS patients was extracted from the peripheral blood of each 20ml, and the peripheral blood mononuclear cells (Peripheral blood mononuclear cells, PBMCs) were extracted and inoculated on the 96 orifice. Results: in 1 healthy subjects, the proportion of MDSCs in peripheral blood (0.65% + 0.28%) was significantly lower than that in MDS (2.39% + 0.74%) (2.39% + 0.74%), and the proportion of MDSCs in peripheral blood (0.65% + 0.28%) in healthy subjects was lower than that of low risk - I MDS (1.46% + 0.17%) (P0.05), and the proportion of MDSCs in peripheral blood (1.46% +) in patients with low and middle risk MDS was lower than that of middle risk - II. The percentage of peripheral blood NK cells in peripheral blood (12.88% + 4.51%) of peripheral blood (12.88% + 4.51%) in patients with high risk MDS (P0.05).2 health was significantly higher than that of MDS (9.08% + 3.59%) (P0.05), and the percentage of NK cells in peripheral blood (12.88% + 4.51%) in healthy subjects was not statistically significant (P0.05) compared with those of low risk - I MDS patients (13.71% + 2.46%), and low endanger. The percentage of NK cells in peripheral blood (13.71% + 2.46%) of peripheral blood (13.71% + 2.46%) in patients with middle risk I was higher than that in middle risk - II and high risk MDS patients (7.53% + 2.36%) (P0.05).3 healthy persons (0.03% + 0.01%) in peripheral blood (0.03% + 0.01%) were significantly lower than those of MDS patients (3.75% + 3.12%) (P0.05), and the percentage of peripheral blood CD34+ cells (0.03% + 0.01%) in healthy subjects was lower than that of low endanger. Risk I MDS patients (0.34% + 0.36%) (P0.05); low risk middle risk - I MDS patients (0.34% + 0.36%) were lower than middle risk - II and high risk MDS patients (4.88% + 2.77%) (4.88% + 2.77%) (P0.05).4 different concentrations of MDSCs, NK cells, CD34+ cells of the proliferation and apoptosis of MDS (P0.05). Conclusion: 1 patients of peripheral blood circulation in the peripheral circulation The proportion of 4+ cells increased and increased with the increase of the risk of disease; the proportion of NK cells was reduced, and with the increase of the risk of disease,.2 ART had no significant effect on the proliferation and apoptosis of MDS peripheral blood of MDSCs, NK cells, CD34+ cells.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R551.3

【參考文獻】

相關(guān)期刊論文 前10條

1 尹t;王春艷;黃敏;毛霞;徐金環(huán);周劍峰;張義成;;髓系來源抑制細胞在預測allo-HSCT后急性移植物抗宿主病中的作用[J];中華器官移植雜志;2016年05期

2 王穎;韓玉祥;喬淑凱;牛志云;潘];;Fe_3O_4磁性納米粒子增強青蒿琥酯誘導SKM-1細胞凋亡(英文)[J];中國實驗血液學雜志;2015年06期

3 劉永華;王朋;江錦紅;章俏雷;曲志剛;馬光麗;汪笑秋;王曉麗;曾玉曉;方炳木;;急性髓系白血病患者外周血表達髓系抑制細胞的臨床研究[J];臨床血液學雜志;2015年06期

4 袁朝暉;沈嬋娟;劉玉霞;鐘永;;多發(fā)性骨髓瘤患者外周血髓系來源的抑制性細胞的分布及臨床意義[J];醫(yī)學臨床研究;2015年05期

5 席微波;王叨;劉玉峰;;兒童骨髓增生異常綜合征28例臨床分析[J];中國當代兒科雜志;2013年07期

6 張秋榮;陳令松;張桂華;徐金格;曹若男;宋文煒;;骨髓增生異常綜合征患者細胞及體液免疫功能分析[J];臨床薈萃;2012年01期

7 王素云;楊曉陽;鄧凱;成志勇;陳浩;楊靜慈;尚銀濤;潘];;青蒿琥酯對骨髓瘤RPMI 8226細胞增殖、凋亡及對Survivin、Caspase-3、Caspase-7的影響[J];中草藥;2010年05期

8 胡守友;朱學軍;范振芳;孔祥圖;陳玉超;陳健一;季建敏;孫雪梅;;吉西他濱化療聯(lián)合樹突狀細胞瘤內(nèi)注射對小鼠巨大淋巴瘤的治療作用[J];中國腫瘤生物治療雜志;2009年06期

9 徐述;許勇鋼;楊曉紅;胡曉梅;王洪志;劉鋒;麻柔;;骨髓增生異常綜合征患者先天性免疫細胞數(shù)量和功能分析[J];中國免疫學雜志;2009年07期

10 謝紅;陳立軍;姚麗;靳秋月;呼文亮;;雙氫青蒿素誘導人腫瘤細胞凋亡及其分子機制的研究[J];中國藥房;2007年24期

相關(guān)博士學位論文 前2條

1 江匯涓;MDSC在骨髓增生異常綜合征中免疫抑制作用的研究[D];天津醫(yī)科大學;2013年

2 王穎;青蒿琥酯抗骨髓增生異常綜合征作用和機制的體外實驗研究[D];河北醫(yī)科大學;2012年

，

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