本文選題：骨髓增生異常綜合征 + 髓樣抑制細(xì)胞�。� 參考：《河北醫(yī)科大學(xué)》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(xì)胞的克隆性疾病,以一系或多系外周血細(xì)胞減少、骨髓無(wú)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(diǎn)。MDS的骨髓微環(huán)境是MDS形成和發(fā)展的關(guān)鍵機(jī)制,腫瘤細(xì)胞通過(guò)利用和改變微環(huán)境來(lái)逃避免疫殺傷而優(yōu)勢(shì)于正常細(xì)胞生長(zhǎng)。髓樣抑制細(xì)胞(Myeloid-derived suppressor cells,MDSCs)是一群具有相似生物學(xué)功能的異質(zhì)性細(xì)胞,包括骨髓祖細(xì)胞、未成熟髓系細(xì)胞(Immature myeloid cells,IMCs)、未成熟的巨噬細(xì)胞和未成熟的樹(shù)突狀細(xì)胞。MDSCs可通過(guò)調(diào)節(jié)固有免疫及適應(yīng)性免疫應(yīng)答促進(jìn)腫瘤發(fā)生免疫逃逸,如抑制CD4+T細(xì)胞、CD8+T細(xì)胞和自然殺傷細(xì)胞(Natural kill cells,NK)等的活化及免疫應(yīng)答。腫瘤微環(huán)境和MDSCs誘導(dǎo)的免疫耐受相互作用來(lái)促進(jìn)MDS的發(fā)展。因MDS多見(jiàn)于老年人,所以不能承受強(qiáng)烈化療及行異基因造血干細(xì)胞移植(Allogeneic hematopoietic stem cell transplantation,Allo-HSCT),尤其對(duì)于高危的MDS患者,極易向白血病轉(zhuǎn)化,故預(yù)后極差。對(duì)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㈣F螯合劑及抗血管增殖藥物等。盡管這些藥物已經(jīng)應(yīng)用于臨床治療MDS,但這些藥物都沒(méi)有獲得持久療效。為中高�；颊邔ふ业投�、有效、經(jīng)濟(jì)的藥物治療是十分需要解決的臨床問(wèn)題。青蒿琥酯(Artesunate,ART)是一種從中藥青蒿中提取出來(lái)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(dǎo)MDS細(xì)胞系SKM-1細(xì)胞凋亡,并且ART與傳統(tǒng)化療藥物的殺傷機(jī)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(shì),如價(jià)格低廉,高效低毒、腫瘤靶向性等。本實(shí)驗(yàn)通過(guò)研究MDS患者外周血MDSCs、CD34+細(xì)胞、NK細(xì)胞數(shù)目以及與MDS疾病惡性程度之間的關(guān)系。通過(guò)體外細(xì)胞培養(yǎng)實(shí)驗(yàn),檢測(cè)ART對(duì)MDS來(lái)源的MDSCs、NK細(xì)胞、CD34+細(xì)胞的各細(xì)胞群的變化,對(duì)進(jìn)一步研究MDS的發(fā)病機(jī)制,ART抗MDS機(jī)制有十分重要的臨床意義,對(duì)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(xué)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(shí)驗(yàn)對(duì)象。同時(shí)將MDS患者行IPSS評(píng)分對(duì)疾病惡性程度進(jìn)行分層分析。取實(shí)驗(yàn)對(duì)象外周血各2ml,采用流式細(xì)胞術(shù)(Flow cytometry,FCM)檢測(cè)其外周血中MDSCs(CD33+CD11b+CD14-HLA-DR-)、NK細(xì)胞(CD3-CD56+)和CD34+細(xì)胞比例。抽取3例MDS患者外周血各20ml,提取外周血單個(gè)核細(xì)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(yīng)各孔48小時(shí),用FCM檢測(cè)MDSCs、NK細(xì)胞、CD34+各群細(xì)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(xì)胞占淋巴細(xì)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(xì)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(wú)統(tǒng)計(jì)學(xué)意義(P0.05);低危及中危-ⅠMDS患者外周血NK細(xì)胞占淋巴細(xì)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(xì)胞百分?jǐn)?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(xì)胞百分?jǐn)?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(duì)MDS外周血來(lái)源的MDSCs、NK細(xì)胞、CD34+細(xì)胞的增殖和凋亡均未見(jiàn)明顯差異(P0.05)。結(jié)論:1 MDS患者外周血循環(huán)中MDSCs和CD34+細(xì)胞比例均是升高的,且隨著疾病危險(xiǎn)程度增加而增高;其N(xiāo)K細(xì)胞比例是降低的,并隨著疾病危險(xiǎn)程度增加而降低。2 ART對(duì)MDS外周血來(lái)源的MDSCs、NK細(xì)胞、CD34+細(xì)胞的增殖和凋亡暫未見(jiàn)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(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R551.3

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