精原干细胞移植后的体内迁移、增殖和分化

背景:精原干细胞移植对男性不育的治疗具有潜在的临床应用价值,但移植后干细胞体内迁移、增殖、分化的过程目前尚不完全清楚.目的:观测精原干细胞移植后的体内迁移、增殖和分化过程.方法:以出生后6～10 d的雄性C57BL/6小鼠为供体,通过复合酶消化、差速贴壁结合非连续性Percoll密度梯度离心的方法获取精原干细胞;以出生后6周的雄性C57BL/6小鼠为受体,腹腔注射白消安,破坏其内源性生精功能.实验组采用曲细精管微注射法将供体精原干细胞移植入受体睾丸内,对移植后细胞进行PKH26-GL荧光追踪分析,观察其体内迁移过程,以Western Blot和RFQ-PCR法检测睾丸组织α6-Integrin,c-kit,SCF蛋白及mRNA的变化.以未接受化疗和细胞移植的正常同系生小鼠作为阳性对照组,以单侧睾丸曲细精管微注射移植细胞递质作为阴性对照组.结果与结论:PKH26-GL荧光追踪移植细胞,移植后1周部分精原干细胞已向曲细精管基底膜迁移,移植后1个月精原干细胞已从曲细精管管腔迁移至曲细精管基底膜,并分裂增殖,移植后3个月曲细精管管腔内可见大量精子细胞形成.移植后1,2,3个月,各组α6-Integrin,c-kit蛋白表达均呈增加趋势(P < 0.01),阴性对照组、实验组SCF蛋白表达有增加趋势(P < 0.05);各组α6-Integrin,c-kit,SCF mRNA的表达均有增加趋势(P < 0.05).提示大剂量化疗后,生精上皮内仍存在一定数量的Sertoli细胞,这种精子发生的微环境并未完全破坏,外源性精原干细胞移植后能在受体Sertoli细胞所提供的微环境中增殖和分化.     

精原干细胞和睾丸组织的同种异体移植

背景:精原干细胞作为精子发生过程的基础和前提,其自我更新和分化途径目前仍不完全清楚。目的:观察非免疫缺陷动物新生Wistar大鼠的精原干细胞和睾丸组织移植于去势成年Wistar大鼠后的成活及生长发育情况。设计、时间及地点:组织细胞形态学水平的随机动物对照实验,于2007-04/08在广西医科大学实验动物中心外科实验室完成。材料:选用健康新生7～9d雄性Wistar大鼠为供体,受体为经过严格检疫合格的8～12周的成年雄性Wistar大鼠,体质量180～220g。方法:取新生雄性大鼠睾丸,采用两步法组合酶顺序消化制备大鼠精原干细胞悬液,以Percoll不连续密度梯度离心法初步纯化精原干细胞。取10只成年雄性大鼠,切除双侧睾丸形成去势大鼠,按随机数字表法分为2组,每组5只。精原干细胞悬液移植组将制取好的1mL精原干细胞悬液在5min内注射至受体背部皮下。睾丸组织块移植组将制备好的已剖开的睾丸组织植入受体背部皮下,每个受体移植2个睾丸4块睾丸组织。主要观察指标:移植物的生长发育情况,移植8周末移植物的组织学特点及C-kit免疫组化定性分析结果。结果:精原干细胞悬液移植后,移植物未见成活生长。睾丸组织块移植后移植物部分...     

一种简便的外周血干细胞冷冻保存法

为了简化传统的细胞冷冻方法。即用10%二甲基亚砜（DMSO）作为冷冻保护剂并用程控降温仪-196℃液氮保存,研究了用5%DMSO,3%羟乙基淀粉（HES）和4%人血白蛋白（HSA）作为冷冻保护剂,而不用程控降温仪直接-80℃冰箱冷冻细胞的方法。对比了不同方法冷冻保存外周血干细胞（PBSCs)的细胞活性,单个核细胞数,CFU-GM和CD34^ 细胞回收率。结果发现,简便冷冻法可获得更高的单个核细胞（MNC）和CFU-GM回收率,无凝集现象;并且经长达24个月冷冻保存后仍可获得满意的CFU-GM和CD34^ 细胞回收率,37℃融冻和20℃室温融冻法之间各项指标无明显差别。细胞融冻后在室温下暴露于5%DMSO中1小时,CFU-GM回收率并不降低。而细胞活性从93.2%降至84.5%,所以,这种简便的细胞冷冻保存法简便易行。可替代传统的液氮保存法,对于没有程控降温仪的单位具有实用价值。而且还可减少病人DMSO输入量第一线决策。     

非程控-80℃冷冻保存自体外周血干细胞移植的临床研究

外周血干细胞(PBSC)大容量单采和非程控-80℃冷冻保存是一种简便、实用的造血干细胞采集和保存方法,国外自90年代以来开始有这方面的报道[1～3],笔者用-80℃冷冻保存的自体PBSC进行移植治疗恶性血液病和实体瘤29例获得成功,现报道如下.     

小鼠精原干细胞染色体G带显示及其影响因素

背景:精原干细胞技术的快速发展为生殖工程带来了新希望,而在体外维持染色体数目及结构的稳定性是其使用的关键问题之一。目的:探索精原干细胞染色体G带显示技术及其影响因素,为如何鉴定培养状态下干细胞的染色体核型提供实验参考。设计:观察实验。单位:泸州医学院材料:实验于2004-03/2005-04在泸州医学院医学分子生物学实验室完成。生后10d左右的昆明系小白鼠(雌雄均可)由泸州医学院动物科提供(许可证号:川实动管质第17号),低糖DMEM,10mg/L丝裂霉素C,IMDM培养基,1×10-5mol/L秋水仙素(磷酸缓冲液配置),7.5mmol/L低渗氯化钾,甲醇:冰醋酸(3∶1)固定液,Giemsa染液,0.25%胰蛋白酶。方法:取小白鼠股骨骨髓,进行饲养层细胞的制备。取生后7～8d左右雄性小白鼠睾丸并制成细胞悬液,调整细胞密度为3×105L-1,接种到预先制备好的骨髓基质细胞饲养层上,细胞在37℃、体积分数0.05的CO2、70%湿度的二氧化碳培养箱中培养。待细胞生长到15～20d时,吸出克隆细胞团,吹打分散后滴加秋水仙素处理4～6h。收集细胞悬液,再经低渗处理后滴片染色。选择染色体分散良好,无重叠的分散中期的细胞进行记数,观察染色体形态。主要观察指标:骨髓基质及精原干细胞的培养及染色体显色与计数。结果:精原干细胞的核型与正常小鼠体细胞的染色体形态一致,染色体呈粒状、棒状,核型为20对,40条。在油镜下镜检可见3种染色体形态,一种呈高浓缩状态的染色体,可计数染色体总数,但不能显出带纹;第2种是已完全展开并铺于赤道板上的处于分裂中期的染色体,染色体总数和带纹显示都非常清楚;第3种是染色体已折转并向两极移动,并逐渐开始浓缩,染色体总数可以计数,带纹不能清楚显示。结论:小鼠精原干细胞染色体受到细胞所处的分裂时期、低渗处理的效果、滴片时细胞的分散程度及胰酶浓度及消化时间等因素的影响。     

大剂量米托蒽醌为主的预处理后非冷冻保存自体干细胞移植治疗晚期淋巴瘤

自体造血干细胞移植能明显改善淋巴瘤患者的生存。在淋巴瘤诱导缓解的化疗方案中，蒽环或蒽醌类药物最重要，但由于其对心脏和肝脏的不良反应，不能提高剂量引入造血干细胞移植的预处理方案，在缓解期、非冷冻自体干细胞支持下单次大剂量应用米托蒽醌还未见报道。我们把对心脏、肝脏不良反应小的米托蒽醌引入预处理方案，并大剂量1次应用，取得显著疗效，7年无病生存率明显提高，并预计可治愈部分淋巴瘤患者，未见明显心、肝、肾不良反应和造血功能减退。     

枸杞多糖对精原干细胞体外增殖的影响

背景：精原干细胞移植对不育具有潜在的临床应用价值,体外建立精原干细胞的培养系统获得数量较多的精原干细胞,仍是目前研究中亟待解决的问题。目的：观察枸杞多糖对精原干细胞体外增殖的影响。方法：采用两步酶消化法获取出生4～6d雄性C57BL/6小鼠睾丸Sertoli细胞与精原干细胞,将精原干细胞接种在Sertoli细胞饲养层上,再加入枸杞多糖或联合细胞因子添加到细胞培养液中。1周后以流式细胞仪检测细胞周期及细胞活性率,并检测各组精原干细胞GFRa-1、Thy-1、c-kit的阳性率。结果与结论：单独加入枸杞多糖后精原干细胞数量明显增加,增殖明显,联合加入胶质细胞源性神经营养因子与白血病抑制因子精原干细胞增殖更加明显（P〈0.05）。并发现体外培养1周后的精原干细胞仍保持其睾丸组织内的精原干细胞特征,大多仍维持在未分化状态。表明在枸杞多糖或枸杞多糖联合胶质细胞源性神经营养因子及白血病抑制因子作用下,可促进精原干细胞体外增殖。     

枸杞多糖对精原干细胞体外增殖的影响

背景:精原干细胞移植对不育具有潜在的临床应用价值,体外建立精原干细胞的培养系统获得数量较多的精原干细胞,仍是目前研究中亟待解决的问题。目的:观察枸杞多糖对精原干细胞体外增殖的影响。方法:采用两步酶消化法获取出生4～6d雄性C57BL/6小鼠睾丸Sertoli细胞与精原干细胞,将精原干细胞接种在Sertoli细胞饲养层上,再加入枸杞多糖或联合细胞因子添加到细胞培养液中。1周后以流式细胞仪检测细胞周期及细胞活性率,并检测各组精原干细胞GFRa-1、Thy-1、c-kit的阳性率。结果与结论:单独加入枸杞多糖后精原干细胞数量明显增加,增殖明显,联合加入胶质细胞源性神经营养因子与白血病抑制因子精原干细胞增殖更加明显(P<0.05)。并发现体外培养1周后的精原干细胞仍保持其睾丸组织内的精原干细胞特征,大多仍维持在未分化状态。表明在枸杞多糖或枸杞多糖联合胶质细胞源性神经营养因子及白血病抑制因子作用下,可促进精原干细胞体外增殖。     

精原干细胞在成骨培养条件下的生物学特征

目的:观察小鼠精原干细胞在成骨培养条件下的生物学特征,为探讨精原干细胞的多向分化能力提供实验依据。方法:实验于2006-03/2007-01在泸州医学院医学分子生物学实验室完成。①实验材料:生后6～8d左右昆明系小白鼠,雌雄均可,由泸州医学院动物科提供。②实验方法:取7～10d小白鼠双侧股骨和胫骨,制备骨髓基质饲养层。取6～8d雄性小鼠,脱颈椎处死后,无菌取出睾丸,进行精原干细胞分离、培养与鉴定。取培养3d的精原干细胞,分为两组:对照组用基本培养液(IMDM 100U/mL青霉素 100U/mL链霉素 10%胎牛血清)培养,实验组用条件培养液培养(基本培养液中添加50mmol/Lβ-甘油磷酸钠、10-6mol/L地塞米松、50mg/L维生素C、50nmol/L胰岛素、20nmol/L碱性成纤维细胞生长因子)培养。③实验评估:在倒置显微镜下观察细胞的形态学变化、紫外分光光度仪动态检测上清液中碱性磷酸酶活性变化、免疫荧光法检测细胞内Ⅰ型胶原的表达。结果:①细胞形态学特征变化:实验组精原干细胞在成骨条件培养液中较对照组细胞贴壁生长快,条件培养3～6d后见细胞生长迅速,呈集落样增长,细胞立体感增强,但未见明显细胞间桥;诱导培养15d见细胞增殖达高峰,增殖细胞的胞质间桥仍不明显。②Ⅰ型胶原免疫荧光检测结果:实验组细胞浆内出现绿色荧光,呈阳性反应,对照组细胞呈阴性反应。③培养上清液碱性磷酸酶活性变化:实验组与对照组培养上清液中的碱性磷酸酶活性开始很低,以后逐渐增强。实验组培养上清液的碱性磷酸酶活性每个时间段都明显高于对照组(P<0.05)。结论:小鼠精原干细胞在成骨诱导培养条件下能快速增殖,增殖过程中表现出成骨细胞的某些生物学特征,这为进一步研究精原干细胞的多向分化潜能提供实验参考。     

以骨髓基质细胞为饲养层培养小鼠精原干细胞

目的:建立精原干细胞与骨髓基质细胞共培养细胞模型,探讨骨髓基质细胞层支持精原干细胞扩增的可能机制。方法:实验于2004-03/2005-04在泸州医学院医学分子生物学实验室完成。以6～8d龄小鼠为材料,脱颈处死后经体积分数为0.75的乙醇浸泡消毒,打开腹腔分离双侧睾丸,采用0.25%胰酶消化离心后培养睾丸细胞,采用差速贴壁法分离支持细胞与生精细胞。同时取小鼠胫骨骨髓,分离后用IMDM培养基培养,待细胞生长至单层时,以10mg/L丝裂霉素C处理的原代培养骨髓基质细胞作为饲养层,并将分离后的生精细胞接种至饲养层上,以含体积分数为0.1的小牛血清的IMDM为培养基,37℃下共培养,在倒置相差显微镜下观察细胞的生长特点,并对培养15d的细胞进行苏木精-伊红染色、碱性磷酸酶染色及C-Kit受体检测。结果:①支持细胞在培养后4～6h就开始贴附变形,此时生精细胞仍呈圆形,且悬浮于培养基中,将悬浮的圆形生精细胞吸出后接种于骨髓基质饲养层上共培养。培养8h后,见体积较小的原形细胞开始贴附增殖,继续培养三四天后见体积相对较大的圆形细胞开始死亡,推测增殖较小的细胞为精原干细胞。增殖细胞的生长特点及形态学特征与文献报道精原干细胞的特征相符合。②苏木精-伊红染色显示培养的生殖细胞大小均匀,核大而圆着色深、胞质浅染、核质比高;碱性磷酸酶染色见胞质和细胞膜为阳性,C-Kit受体显示细胞膜呈强阳性,背景基质细胞为阴性。这些均符合精原干细胞的生物学特征。结论:精原干细胞在未加入细胞因子、原代培养的骨髓基质细胞饲养层中能较长时间生长并分裂增殖,推测骨髓基质细胞通过分泌生物活性物质和细胞黏附分子支持精原干细胞的扩增。     

Hematopoietic stem cell processing and cryopreservation

Either bone marrow or peripheral blood may be harvested to provide hematopoietic stem cells (HSC) for autologous transplantation. Both, however, comprise heterogeneous cell populations. The HSC necessary for successful engraftment constitute a very small fraction of the cells harvested. After collection, the harvested cells usually undergo several processing steps to reduce the product volume, remove cells (such as mature blood cells or tumor cells), or to cryopreserve the cells for later reinfusion. Granulocytes and red blood cells, for example, survive cryopreservation poorly using freezing techniques designed for HSC. Therefore, bone marrows being cryopreserved must be depleted of mature blood cells to avoid toxicity from infusion of damaged mature blood cells. Mature blood cells may also impede the variety of tumor cell purging techniques currently being studied. These processings are designed to minimize the loss of HSC while achieving an appropriate HSC product for the individual patient. A number of apheresis devices and cell washers simplify the enrichment of HSC in the harvested cell products. In contrast, tumor cell purging techniques are not standardized between the various transplant centers. © 1992 Wiley-Liss, Inc.     

人脐带间充质干细胞冻存复苏后的生物学特征

背景:冻存脐带间充质干细胞,并有效地保持其生物学特性,是储存脐带间充质干细胞以供临床使用的重要工艺之一.目的:观察冻存后脐带间充质干细胞的生物学特性,验证其是否仍具有间充质干细胞的基本特征.方法:从脐带分离得到间充质干细胞后,将其冻存于液氮之中.比较经冻存复苏后的脐带间充质干细胞和新鲜制各的脐带间充质干细胞活率、抑制人外周血单个核细胞分泌γ-干扰素等方面的异同.检验经冻存复苏后的脐带间充质干细胞是否具有多向分化潜能,其表型是否满足间充质干细胞的基本特征.结果与结论:在细胞活率和抑制人外周血单个核细胞分泌γ-干扰紊方面,冻存和新鲜的脐带间充质干细胞差异无显著性意义.经冻存复苏后的脐带间充质干细胞保持了间充质干细胞的基本形态,其表型满足间充质干细胞的基本要求,具有多向分化的潜能.     

A theoretically optimized method for cord blood stem cell cryopreservation

The objective of this study was to develop an optimal cryopreservation method for human umbilical cord blood hematopoietic progenitor cells as evidenced by improved retention of in vivo engraftment ability and multilineage differentiation. An extended understanding of the osmometric/permeability characteristics of cord blood stem cells was accomplished by measuring permeability of the cryoprotectant dimethyl sulfoxide (DMSO) at below-ambient temperatures (10 degrees and 3 degrees C). These data were combined with previously published osmotic and permeability data and the water-NaCl-DMSO phase diagram in conjunction with a mathematical model to determine an optimal initial DMSO concentration, cooling rate, and liquid nitrogen plunging temperature. Cells cryopreserved with the theoretically optimized procedure were then compared with cells frozen using standard methods for the ability to engraft in irradiated NOD/SCID mice. The optimal procedure was determined to include a 0.7 molal (approximately 5%) DMSO concentration at a cooling rate of 4 degrees C/min, and a plunging temperature of -44 degrees C. The optimized protocol resulted in significantly higher engraftment of human CD45(+) cells (17.2 +/- 1.6% vs. 8.4 +/- 1.6%), CD19(+) B lymphocytes (11.3 +/- 1.2% vs. 5.8 +/- 1.2%), and CD34(+) cells (1.9 +/- 0.09% vs. 0.6 +/- 0.09%) compared to cells frozen using a standard method. Engraftment of CD33(+) cells was not significantly different (4.0 +/- 0.3 vs. 3.2 +/- 0.6, respectively). This study demonstrated that the use of a theoretically determined optimal cryopreservation method is superior to standard methods for maintaining UCB PCBs with multilineage repopulation potential in NOD/SCID mice.     

Hematopoietic stem cell cryopreservation: a review of current techniques.

Hematopoietic stem cells (HSC) can be stored for prolonged periods at cryogenic temperatures. The techniques currently used were derived from the initial report in 1949 of cryopreservation of bovine sperm in glycerol. The addition of this penetrating cryoprotectant protected the cells from the injury associated with ice formation. Current cryopreservation techniques (with minor variations) suspend cells in an aqueous solution of salts, protein, and one or more cryoprotectants. Cells are frozen at slow rates and stored generally below -120 degrees C in mechanical freezers or nitrogen refrigerators. That these techniques are successful in maintaining HSC viability is evident from the engraftment of these cells in patients treated with marrow-lethal conditioning regimens. However, issues such as the composition of the cryoprotectant solution, cell concentration during freezing, cryoprotectant toxicity, and storage temperatures have not been adequately studied, primarily because of a lack of appropriate assays for HSC cryosurvival. HSC cryobiology will become an increasingly important subject as new HSC collection and processing techniques are developed. Improved cryosurvival of HSC using modified cryoprotectant solutions may improve engraftment kinetics and decrease the cost and morbidity of autologous transplantation.     

Effect of nucleated cell count and cryopreservation on engraftment post autologous stem cell transplant

Using 753 collections from 426 adult haematology patients, we conducted a retrospective, analysis into the effects of overnight storage and nucleated cell counts (NCC) on viable, CD34⁺ (vCD34⁺) recovery and engraftment kinetics post autologous stem cell, transplant (ASCT) with peripheral blood stem cells (PBSC). There were significant, differences in vCD34 + recovery ( P < 0.01) after cryopreservation associated with, the fresh NCC of ≥ 300 × 10 ⁶ /mL in products stored overnight, but no association, with time to platelet or neutrophil engraftment post-ASCT was observed for these, products. There was no association of vCD34⁺ numbers or engraftment kinetics with cryopreserved NCC with either below or greater than the local recommended concentration of 400 × 10⁶ /mL of product. However, there was significant difference in engraftment kinetics in relation to the viable CD34⁺ dose given at ASCT, in relation to the time to early engraftment and the amount of platelet support given during the engraftment period post-ASCT. We conclude the vCD34⁺ dose at ASCT is of great importance to early engraftment kinetics and that NCC is an important factor during overnight storage, but not for cryopreservation of PBSC. In light of our findings, we recommend that apheresis products collected in a closed system can safely be stored undiluted overnight.     

Parameters of cell freezing: Implications for the cryopreservation of stem cells

The overall objective of this review has been to discuss specific parameters that may influence the ability to successfully cryopreserve stem cells and, more importantly, stem-cell-based therapies. This discussion of factors is in no way complete. Specifically, the effect of temperature and the duration of storage, sensitivities to cryopreservation of bone marrow cells from patients with specific disorders (for example, chronic myelogenous leukemia), and the postfreeze processing of cells are factors of clinical significance that, for the sake of brevity, have been omitted.As new stem-cell-based therapics (gene, stem cell transplant, or immunotherapy) become the standard of care for a wide variety of diseases, appropriate cryopreservation protocols will be necessary to increase patient access, reduce cost, and enhance the safety and effectiveness of these therapies. Appropriate protocols must include methods and reagents appropriate for human use. The cryopreservation protocols developed must also reflect the biological and physical properties of the cells that can be altered significantly by the culture process. Finally, cryopreservation studies should be performed concurrently with in vitro culture studies to reduce the overall cost and time required for the development or validation of a cryopreservation protocol.     

Thrombotic microangiopathy following stem cell transplantation

Stem cell transplantation associated microangiopathy (TA-TMA) occurs in about 7% of all transplantations. While it resembles in all its features idiopathic TTP or HUS, response to plasma therapy is poor. The overall mortality of patients afflicted with TA-TMA is about 80%. Comparative studies aimed at discovering risk factors for the development of TA-TMA and defined different entities according to clinical, laboratory and outcome parameters. It is thought to be of multifactorial aetiology with the central event of endothelial damage. Intensive chemotherapy and radiation for bmt-preparation, infection in the immunocompromised host and graft versus host disease in conjunction with immunosuppressive therapy i. e. cyclosporine turn the haemostatic balance into a procoagulant state. Therapeutic strategies deduced from idiopathic TTP were disappointing with the only consensus in the withdrawal of cyclosporine from TA-TMA patients. Future trials are directed to ameliorate endothelial dysfunction.     

药物去势与异基因造血干细胞移植后胸腺损伤关系的实验研究

目的 探讨促黄体激素释放激素(LHRH)去势对异基因造血干细胞移植(allo-HSCT)后小鼠胸腺功能的保护作用.方法 构建C57BL/6→BALB/c小鼠MHC不相合allo-HSCT模型;对急性移植物抗宿主病(aGVHD)严重程度评分;采用蛋白液相芯片技术榆测小鼠胸腺细胞内IFNγ、TNFα和IL-1β表达水平,实时定量PCR分析小鼠外周血T细胞受体重排删除环(sjTREC)水平.结果 单纯allo-HSCT组(A组)、LHRH处理allo-HSCT组(B组)、同基因移植组(C组)三组小鼠均获得造血重建,WBC>1.0×109/L时间分别为(11.2±1.4)、(9.8±0.6)和(9.7±0.7)d(P=0.003).A、B两组aGVHD出现时间分别为(11.4±1.2)d和(14.1±0.7)d(P=0.000),C组未发生aGVHD,A、B组aGVHD发生率为100%.A组和B组aGVHD评分分别为(9.1±0.7)和(5.1±1.0)分(P=0.000).正常受鼠胸腺细胞内IFNγ、TNFα和IL-1β水平分别为(2.3±2.5)、(1.7±1.1)和(1.8±1.2)pg/ml.A、B、C三组IFNγ水平分别为(10.5±2.1)、(6.7±2.1)和(5.2±3.3)ng/ml;TNFα水平分别为(7.0±2.6)、(4.3±0.8)和(3.0±1.8)pg/ml;IL-1β水平分别为(24.9±9.0)、(17.4±3.9)和(10.8±3.1)pg/ml.A、B组IFNγ、TNFα和IL-1β水平比正常受鼠组明显升高(P值均<0.01);C组IFNγ和IL-1β水平明显高于正常对照组(P值分别为0.015,0.013).A组IFNγ、TNFα和IL-1β水平比B组明显升高(P值分别为0.002、0.002、0.004);A组IFNγ、TNFα和IL-1β水平较C组明显升高(P值均为0.000).Linear Regression分析提示,IFNγ和TNFα水平越高,aGVHD评分越高(r2=0.359,P=0.05;r2=0.228,P=0.019).正常受鼠1000个外周血单个核细胞中sjTREC拷贝数为(39.4±44.7).A、B和C组小鼠分别为(12.3±13.0)、(58.0±71.8)和(19.6±14.6),与正常受鼠组sjTRECs水平差异无统计学意义.结论 细胞因子IFNγ、TNFα和IL-1β参与了aGVHD对胸腺的损伤过程;LHRH药物去势能减轻aGVHD对胸腺的损伤,并通过保护胸腺减轻aGVHD的严重程度,其机制可能与降低胸腺细胞内IFNγ、TNFα水平有关.