Advances and perspectives on cellular therapy in acquired bone marrow failure diseases

Acquired bone marrow failure diseases(ABMFD) are a class of hematopoietic stem cell diseases with a commonality of non-inherited disruption of hematopoiesis that results in pancytopenia. ABMFDs also are a group of heterogeneous diseases with different etiologies and treatment options. The three most common ABMFDs are aplastic anemia, myelodysplastic syndrome, and paroxysmal nocturnal hemoglobinuria. Stem cell transplantation is the only treatment that can cure these diseases. However, due to high therapy-related mortality, stem cell transplantation has rarely been used as a first line treatment in treating ABMFD. With the advance of personalized medicine and precision medicine, various novel cellular therapy strategies are in trial to increase the efficiency and efficacy of ABMFD treatment. This article aims to review current available stem cell transplantation protocols and promising cellular therapy research in treating ABMFD.

Repeated application of autologous bone marrow-derived stem cell therapy in patients with severe Buerger’s disease

Stem cell therapy (SCT) is a promising and prospective approach in the treatment of patients with severe peripheral arterial disorders, primarily with Buerger’s disease. However, very little is known about the duration of the effect of SCT, and to our best knowledge no data are available on the efficacy and safety of repeated SCT in patients with Buerger’s disease. Here we report on two patients with severe Buerger’s disease, who received repeated autologous bone marrow-derived stem cell therapy. Our results show that a second SCT, applied to a previously treated leg 30 or 36 months after the first treatment was efficient in both cases. After twelve months, the clinical state of the repeatedly treated lower limb improved spectacularly and non-healing ulcers healed more rapidly than after the first SCT. No severe adverse events were detected. Thus repeated SCT offers a safe and efficient treatment option for relapsing patients at the advanced stage of Buerger’s disease.

Overexpression of lentivirus-mediated glial cell line-derived neurotrophic factor in bone marrow stromal cells and its neuroprotection for the PC12 cells damaged by lactacystin

Objective To construct recombinant lentiviral vectors for gene delivery of the glial cell line-derived neurotropnic factor （GDNF）, and evaluate the neuroprotective effect of GDNF on lactacystin-damaged PC12 cells by transfecting it into bone marrow stromal cells （BMSCs）. Methods pLenti6/V5-GDNF plasmid was set up by double restriction enzyme digestion and ligation, and then the plasmid was transformed into Top10 cells. Purified pLenti6/V5-GDNF plasmids from the positive clones and the packaging mixture were cotransfected to the 293FT packaging cell line by Lipofectamine2000 to produce lentivirus, then the concentrated virus was transduced to BMSCs. Overexpression of GDNF in BMSCs was tested by RT-PCR, ELISA and immunocytochemistry, and its neuroprotection for lactacystin-damaged PC12 cells was evaluated by MTT assay. Results Virus stock of GDNF was harvested with the titer of 5.6×10^5 TU/mL. After tmnsduction, GDNF-BMSCs successfully secreted GDNF to supematant with nigher concentration （800 pg/mL） than BMSCs did （less than 100 pg/mL）. The supematant of GDNF-BMSCs could significantly alleviate the damage of PC12 cells induced by lactacystin （10 μmol/L）. Conclusion Overexpression of lentivirus-mediated GDNF in the BMSCs cells can effectively protect PC12 cells from the injury by the proteasome inhibitor.

Multiple myeloma mesenchymal stromal cells: Contribution to myeloma bone disease and therapeutics

Multiple myeloma is a hematological malignancy inwhich clonal plasma cells proliferate and accumulate within the bone marrow. The presence of osteolytic le-sions due to increased osteoclast(OC) activity and sup-pressed osteoblast(OB) function is characteristic of the disease. The bone marrow mesenchymal stromal cells(MSCs) play a critical role in multiple myeloma patho-physiology, greatly promoting the growth, survival, drug resistance and migration of myeloma cells. Here, we specifically discuss on the relative contribution of MSCs to the pathophysiology of osteolytic lesions in light of the current knowledge of the biology of my-eloma bone disease(MBD), together with the reported genomic, functional and gene expression differences between MSCs derived from myeloma patients(pMSCs) and their healthy counterparts(dMSCs). Being MSCs the progenitors of OBs, pMSCs primarily contribute to the pathogenesis of MBD because of their reduced osteogenic potential consequence of multiple OB inhibi-tory factors and direct interactions with myeloma cells in the bone marrow. Importantly, pMSCs also readily contribute to MBD by promoting OC formation and ac-tivity at various levels(i.e., increasing RANKL to OPG expression, augmenting secretion of activin A, uncou-pling ephrinB2-EphB4 signaling, and through augment-ed production of Wnt5a), thus further contributing to OB/OC uncoupling in osteolytic lesions. In this review, we also look over main signaling pathways involved in the osteogenic differentiation of MSCs and/or OB activity, highlighting amenable therapeutic targets; in parallel, the reported activity of bone-anabolic agents(at preclinical or clinical stage) targeting those signaling pathways is commented.

Bone marrow-derived mesenchymal stem cells increase dopamine synthesis in the injured striatum

Previous studies showed that tyrosine hydroxylase or neurturin gene-modified cells transplanted into rats with Parkinson＇s disease significantly improved behavior and increased striatal dopamine content. In the present study, we transplanted tyrosine hydroxylase and neurturin gene-modified bone marrow-derived mesenchymal stem cells into the damaged striatum of Parkinson＇s disease model rats. Several weeks after cell transplantation, in addition to an improvement of motor function tyrosine hydroxylase and neurturin proteins were up-regulated in the injured striatum, and importantly, levels of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid increased significantly. Furthermore, the density of the D2 dopamine receptor in the postsynaptic membranes of dopaminergic neurons was decreased. These results indicate that transplantation of tyrosine hydroxylase and neurturin gene-modified bone marrow-derived mesenchymal stem cells increases dopamine synthesis and significantly improves the behavior of rats with Parkinson＇s disease.

The effectiveness of the mononuclear fraction of autologous bone marrow in the treatment of experimental chronic limb ischemia

Objective To study of the effectiveness of using autologous mononuclear fraction of bone marrow for the treatment of chronic limb ischemia. Methods Results of autologous mononuclear fraction of bone marrow in 90 laboratory Wistar rats on a background of creating chronic limb ischemia was presented. Sampling was carried out from the bone marrow of the femur of the animal. The mononuclear fraction of bone marrow autologous 4 × 106 cells in a volume of 200 microliter were injected into the ischemic limb of the two points,in each of which 100 microliter:(1)Paravessel directly below the inguinal ligament at the level of the sacroiliac joint in the area of the anatomical location of collaterals in the projection of the internal iliac artery and its branches;(2)Intramuscularly in gastrocnemius muscle anterior-lateral surface of the middle third of the leg. Results In the experimental group of rats treated with autologous mononuclear fraction of bone marrow,the level of microcirculation compared with the intact group of animals on day 21 was higher than 6. 1% by day 28% ~ 31. 2%; compared with the control group-day 10 increased by 111% at day 21,85. 7% on day 28% ~ 97%. Conclusion Proposed method of treating pathogenically justified and can be recommended for use in clinical practice in the treatment of patients with chronic obliterating diseases of lower limb arteries.

Thiopurine S-methyltransferase polymorphisms and thiopurine toxicity in treatment of inflammatory bowel disease

AIM: To evaluate the relationship between thiopu- rine S-methyltransferase (TPMT) polymorphisms and thiopurine-induced adverse drug reactions (ADRs) in inflammatory bowel disease (IBD). METHODS: Eligible articles that compared the frequency of TPMT polymorphisms among thiopurine-tolerant and-intolerant adult IBD patients were included. Statistical analysis was performed with Review Manager 5.0. Sub-analysis/sensitivity analysis was also performed. RESULTS: Nine studies that investigated a total of 1309 participants met our inclusion criteria. The inci- dence of TPMT gene mutation was increased 2.93-fold (95% CI: 1.68-5.09, P = 0.0001) and 5.93-fold (95% CI: 2.96-11.88, P < 0.00001), respectively, in IBD patients with thiopurine-induced overall ADRs and bone marrow toxicity (BMT), compared with controls. The OR for TPMT gene mutation in IBD patients withthiopurine-induced hepatotoxicity and pancreatitis was 1.51 (95% CI: 0.54-4.19, P = 0.43) and 1.02 (95% CI: 0.26-3.99, P = 0.98) vs controls, respectively. CONCLUSION: This meta-analysis suggests that the TPMT polymorphisms are associated with thiopurine-induced overall ADRs and BMT, but not with hepatotoxicity and pancreatitis.

Imaging of gaucher disease

Gaucher disease is the prototypical lysosomal storage disease.It results from the accumulation of undegrad-ed glucosylceramide in the reticuloendothelial system of the bone marrow,spleen and liver due to deficiency of the enzyme glucocerebrosidase.This leads to he-matologic,visceral and skeletal maifestions.Build up of glucosylceramide in the liver and spleen results in hepatosplenomegaly.The normal bone marrow is re-placed by the accumulating substrate leading to many of the hematologic signs including anemia.The visceral and skeletal manifestations can be visualized with vari-ous imaging modalities including radiography,com-puted tomography,magnetic resonance imaging(MRI)and radionuclide scanning.Prior to the development of enzyme replacement therapy,treatment was only sup-portive.However,once intravenous enzyme replace-ment therapy became available in the 1990s it quickly became the standard of care.Enzyme replacement therapy leads to improvement in all manifestations.Thevisceral and hematologic manifestations respond more quickly usually within a few months or years.The skel-etal manifestations take much longer,usually several years,to show improvement.In recent years newer treatment strategies,such as substrate reduction thera-py,have been under investigation.Imaging plays a key role in both initial diagnosis and routine monitoring of patient on treatment particularly volumetric MRI of the liver and spleen and MRI of the femora for evaluating bone marrow disease burden.

多发性骨髓瘤骨病微环境改变与治疗进展

多发性骨髓瘤骨病(MBD)是多发性骨髓瘤(MM)患者临床常见的并发症,骨相关事件的发生严重影响患者的生存质量与预后。MBD的发病机制涉及生理性骨重塑的失衡,如破骨细胞的过度激活、成骨细胞的抑制及微环境中骨细胞、骨髓基质细胞及免疫细胞等多种细胞与细胞因子的相互作用。目前MBD的治疗在标准抗骨髓瘤治疗基础上,控制肿瘤进展,同时在适应证范围内使用骨相关药物作用于骨重塑。为防止骨破坏,在抗骨吸收的同时,诱导新骨形成来修复现有病变是必不可少的,目前多种新型骨靶向药物在临床前及临床试验中表现出抗骨病作用。本文总结了骨髓微环境改变、骨重塑机制及治疗的新进展。

乙型肝炎肝硬化合并骨质疏松患者临床干预的安全性研究

目的:探究乙型肝炎肝硬化合并骨质疏松患者同时给予抗病毒治疗及抗骨质疏松治疗的安全性.方法:选取我院2020年1月—2022年12月收治的74例乙型肝炎肝硬化合并骨质疏松患者为研究对象,按照随机数字表法将其分为对照组与试验组,每组37例.对照组采用抗病毒药物治疗,试验组在对照组基础上采用抗骨质疏松药物治疗.比较2组患者的肝功能、病毒载量、肝硬度、关键位置骨密度及不良反应发生情况.结果:入院时,入院6、12个月后,2组肝功能、病毒载量、肝硬度情况比较,组间差异无统计学意义(P>0.05).入院6、12个月后,试验组腰椎L_(2~4),股骨颈骨密度均大于对照组,组间差异有统计学意义(P<0.05).2组不良反应发生率比较,差异无统计学意义(P>0.05).结论:同步给予乙型肝炎肝硬化联合骨质疏松患者抗病毒及抗骨质疏松药物治疗安全性较好,增用抗骨质疏松药物在改善患者骨质疏松病情的同时未对患者肝功能产生不利影响.

中药诱导骨髓间充质干细胞的定向分化:研究与进展

背景:近年来,对传统单味中药或提取物、复方中药及含药血清调控骨髓间充质干细胞定向分化为肌成纤维细胞、软骨细胞、成骨细胞、心肌细胞、神经细胞等研究的不断深入,使中药或其提取物对骨髓间充质干细胞增殖、调控定向分化和移植等方面成为组织工程研究领域的一大亮点。目的:综述中药或其提取物诱导骨髓间充质干细胞定向分化的最新研究进展。方法:分别以"中药,定向分化,骨髓间充质干细胞","Chinese herb,directional differentiation,mesenchymal stem cells"为检索词,由第一作者检索2010年1月至2016年1月中国期刊全文数据库、Pub Med数据库及万方数据库相关文章。排除缺乏原创性及重复性研究的文献,计算机初检到99篇文献,最终保留43篇进行归纳总结。结果与结论:骨髓间充质干细胞作为骨分化系统中最强的种子细胞,具有广泛的定向分化潜能,与传统中草药结合在临床治疗众多难治性疾病中凸显重要价值,尤其对于治疗骨代谢疾病、骨缺损、骨折不愈合及迟缓愈合等疾病更加充分发挥其临床应用价值,这既有利于进一步深层次,多角度研究中药的作用及发生机制,也使骨髓间充质干细胞向更宽、更广领域参与临床各类难治性疾病的治疗。

四物汤对血虚证小鼠骨髓细胞CD34抗原表达的影响

目的探讨四物汤对60 Coγ射线或环磷酰胺诱导的血虚证小鼠骨髓细胞CD 34抗原分子表达的影响。方法用60 Coγ射线全身照射小鼠或腹腔注射环磷酰胺造成小鼠血虚证模型 ,实验分正常组、模型组、小剂量组、中剂量组、大剂量组 ,荧光标记小鼠骨髓细胞并用流式细胞仪测定CD 34+ 细胞在骨髓有核细胞中的比例 ,同时做小鼠骨髓细胞集落培养。结果与结论四物汤可促进血虚小鼠骨髓中干祖细胞增生 ,增加CD 34抗原分子的表达 。

芪胶升白胶囊对预防鼻咽癌患者同步放化疗后骨髓抑制疗效观察

目的观察芪胶升白胶囊在鼻咽癌患者同步放化疗所致骨髓抑制中的治疗作用。方法 90例同步放化疗过程中出现骨髓抑制的局部晚期鼻咽癌患者随机分为3组,实验组:芪胶升白胶囊+重组人粒细胞集落刺激因子(recombinant human granulocyte colony-stimulating factor,G-CSF);对照1组:利血生+G-CSF;对照2组:单独使用G-CSF,治疗同步放化疗所致骨髓抑制,比较其疗效及不良反应。结果治疗结束时,实验组与对照1、2组在骨髓保护方面,差异有统计学意义(P<0.05);同步放化疗结束后,及结束后1、2周,实验组较对照1、2组骨髓抑制发生率明显降低,差异均有统计学意义(P<0.05),且无不良反应。结论芪胶升白胶囊对于同步放化疗所致的白细胞下降治疗作用显著,同时可以起到较好的骨髓保护作用,且无明显不良反应,价格低廉,可作为放化疗骨髓抑制时的常规口服用药。

升血方剂对结直肠癌化疗患者免疫功能及骨髓抑制的影响

目的探讨升血方剂对结直肠癌化疗患者免疫功能及骨髓抑制的影响。方法以2011年2月至2015年6月在贵州省中医院肛肠科住院治疗的94例转移性结直肠癌患者为研究对象,根据治疗方案的不同分成观察组和对照组,每组各47例,对照组采用常规的XELON方案进行化疗,观察组在对照组化疗方案的基础上给予升血方剂治疗,治疗两个周期后检测两组治疗后的免疫功能及骨髓抑制情况。结果观察组治疗后稳定率为63.83%,明显高于对照组42.55%(P<0.05);观察组治疗后CD4^+、CD4^+/CD8^+及NK细胞活性明显高于对照组(P<0.05),而对照组治疗后CD4^+、CD4^+/CD8^+及NK细胞活性于治疗前相比差异无统计学意义(P>0.05);观察组治疗后CD4^+、CD4^+/CD8^+及NK细胞活性明显高于对照组治疗后CD4^+、CD4^+/CD8^+及NK细胞活性(P<0.05);观察组治疗后白细胞、中性粒细胞、血小板、血红蛋白减少的发生率分别是70.21%、70.21%、17.02%、14.89%,低于对照组对照组白细胞、中性粒细胞、血小板、血红蛋白减少的发生率,分别是85.11%、87.23%、25.53%、23.40%,观察组白细胞、中性粒细胞减少发生率明显低于对照组(P<0.05);观察组和对照组治疗后生活质量KPS评分均高于治疗前(P<0.05);观察组治疗后生活质量KPS评分明显高于对照组治疗后(P<0.05),观察组G-CSF使用率低于对照组(P<0.05)。结论升血方剂能有效提高晚期转移性结直肠癌患者的免疫功能,减轻骨髓抑制作用,提高生活质量。

乳腺癌骨转移化疗后骨髓抑制的临床分析

目的观察乳腺癌骨转移患者化疗后的骨髓抑制程度。方法收集183例伴有骨转移的晚期乳腺癌患者为研究组,86例晚期无骨转移的乳腺癌患者为对照组。平均化疗2周期后,比较两组的骨髓抑制情况和其它不良反应。结果研究组化疗后白细胞、粒细胞、血小板及红细胞下降Ⅲ～Ⅳ度的发生率分别为46.6%、48.1%、37.2%和24.0%,对照组为31.4%、26.7%、16.3%和11.6%,差异有统计学意义(P<0.05)。研究组FEC/CEF方案白细胞、粒细胞、血小板下降Ⅲ～Ⅳ度的发生率分别为60.8%、62.7%、37.3%,对照组为32.0%、36.0%、12.0%,差异有统计学意义(P<0.05),Ⅲ～Ⅳ度红细胞下降的发生率两组分别为27.5%和8.0%,差异无统计学意义(P>0.05)。AT方案白细胞、粒细胞、血小板和红细胞下降Ⅲ～Ⅳ度的发生率研究组为65.1%、69.8%、36.5%和26.9%,对照组为39.3%、32.1%、14.2%和7.1%,差异有统计学意义(P<0.05)。XT方案白细胞、粒细胞、血小板和红细胞下降Ⅲ～Ⅳ度的发生率研究组为16.7%、14.3%、21.4%和14.3%,对照组为14.3%、9.5%、9.5%和14.3%,差异无统计学意义(P>0.05)。白细胞、血小板、红细胞的骨髓抑制最低点出现时间研究组分别为第8、12、16天,对照组为第12、16、18天,差异有统计学意义(P<0.05)。研究组Ⅲ～Ⅳ度神经毒性发生率为16.9%,对照组为7.0%,差异有统计学意义(P<0.05)。研究组Ⅲ～Ⅳ度感染发生率为9.8%,对照组为2.7%,差异有统计学意义(P<0.05)。结论乳腺癌骨转移患者化疗后总体的骨髓抑制较无骨转移的晚期乳腺癌严重,神经毒性和感染的发生率较高,应注意药物的选择和剂量调整。

恶性婴儿型石骨症相关眼部病变的临床特征分析

背景恶性婴儿型石骨症患者临床上非常少见，合并眼部症状的病例罕见。由于该病延误治疗时预后差，因此受到临床医师的高度关注，而恶性婴儿型石骨症合并眼部症状的患儿治疗后眼部症状是否改善也是值得研究的问题。目的分析恶性婴儿型石骨症合并眼部症状患者的发病特点、眼部表现、治疗及预后。方法收集在首都医科大学附属北京儿童医院确诊的恶性婴儿型石骨症合并眼部症状的患儿2例，对患儿进行全身临床检查和眼部检查，包括一般体格检查、实验室血液学检查、腹部B型超声检查、全身骨骼x线摄片、外眼检查、闪光视觉诱发电位（F—VEP）检查和眼部CT检查。对患儿行骨髓造血干细胞移植术并进行5年的随访，观察患儿的症状改善情况。结果患儿白细胞总数增加、血小板数量减少，进行性贫血，腹部B型超声检查提示肝脏、脾脏肿大，全身骨骼系统均有密度增高、增厚等特征性改变。治疗前1例女性患儿同时存在视神经管狭窄、视盘苍白、右侧面部肌肉麻痹和麻痹性内斜视；而另1例男性患儿存在视神经管狭窄和交替性外斜视，2例患儿均存在F—VEPP，波潜伏期延长和振幅下降。2例患儿均成功进行骨髓造血干细胞移植术，术后全身症状均明显改善，女性患儿F-VEP结果稍有改善，术后1年视神经管直径为1．9mm，随访至5年视神经管直径增宽到3．2mm，但斜视和视神经萎缩症状无改善；而男性患儿除视神经管增宽外，双眼共转性斜视症状消失。结论斜视和眼部症状是恶性婴儿型石骨症重要临床表现的一部分，是中枢神经系统受损的反映，发病机制尚不明确，应该引起重视。恶性婴儿型石骨症的早期治疗可有效改善全身症状和部分眼部症状，挽救患者的视功能。

血液病骨髓移植后卵巢早衰育龄女性激素补充治疗临床疗效

目的:探讨血液病骨髓移植(BMT)后卵巢早衰(POF)患者行激素补充治疗(HRT)的临床疗效。方法:选择2015年4月至2017年3月于苏州大学附属第一医院妇科门诊就诊的血液病BMT后POF行HRT的44例育龄女性的病例资料,比较其HRT前后月经情况、血管舒缩症状、症状自评量表(SCL-90)、卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E_2)、抗苗勒管激素(AMH)、子宫大小、窦卵泡数量(AFC)、乳房钼靶或B超检查以及骨密度的变化情况,初步探讨HRT对这类患者的临床疗效。结果:血液病BMT后发生POF的患者行HRT后,月经复潮率95. 5%,治疗前有血管舒缩症状的患者治疗后症状均缓解(100%),治疗前后症状自评量表(SCL-90)总分和各因子分数差异有统计学意义(P<0. 05)。治疗6个月后AMH数值增加、FSH和LH明显降低、子宫体积增大(P<0. 05),但是AFC无明显变化。治疗1年后复查骨密度的9例患者,L1～L4骨密度无明显变化,双股骨骨密度数值增大(P<0. 05);治疗1年以上的患者有11例随访乳房情况未发现明显变化。结论:血液病BMT后POF的育龄女性行HRT可有效缓解其雌激素缺乏相关症状,建议对这类患者行HRT。

袁氏生脉成骨胶囊对激素诱导骨髓基质细胞成脂分化的拮抗作用

【目的】观察中药袁氏生脉成骨胶囊(主要由丹参、川芎、鸡血藤、黄芪组成)对激素诱导骨髓基质细胞成脂分化的拮抗作用,探讨袁氏生脉成骨胶囊防治激素性股骨头坏死的病理机制。【方法】采用骨髓细胞体外培养技术和血清药理学方法,分离培养大鼠骨髓基质细胞;地塞米松诱导骨髓基质细胞的成脂分化,以含中药血清作为拮抗剂。检测培养细胞的甘油三酯含量和碱性磷酸酶活性。【结果】分离细胞培养4d后可以观察到大量成骨细胞、成纤维细胞、破骨细胞;模型组加入地塞米松造模;成骨胶囊药物组地塞米松用法同模型组,同时加入含药血清。6d后,光镜下观察成骨胶囊药物组见少量脂肪细胞,模型组脂肪细胞明显增多。成骨胶囊药物组碱性磷酸酶活性高于模型组(P<0.05);甘油三酯含量低于模型组(P<0.01)。【结论】中药袁氏生脉成骨胶囊能够有效拮抗激素诱导骨髓基质细胞成脂分化。提示活血化瘀类中药防治股骨头坏死的机理不仅是改善了股骨头的微循环,而且抑制了骨髓基质细胞成脂分化,增加了成骨细胞的表达。

芪胶升白胶囊联合化疗对肿瘤患者白细胞减少及化疗效果的临床观察

目的观察芪胶升白胶囊联合化疗对肿瘤患者白细胞及化疗效果的疗效。方法 83例肿瘤化疗患者随机分为联合组(化疗联合口服芪胶升白胶囊)42例和对照组(化疗)41例。联合组患者口服芪胶升白胶囊2 g/次,3次/d,服用6天为1个疗程,对照组仅予常规化疗,5个疗程后观察检测白细胞水平(WBS)、血红蛋白(Hb)和血小板(Plt)检测值及身体指标,并观察不良反应。结果 5个疗程后,联合组WBC、Hb、Plt均较治疗前升高,而对照组较治疗前降低,其明显低于联合组(P<0.05);通过临床症状观察,联合组缓解率和未进展率分别为59.5%和73.8%,而对照组的缓解率和未进展率分别为41.5%和58.5%,两组比较差异有统计学意义(P<0.05)。结论芪胶升白胶囊对肿瘤患者骨髓功能有保护作用,并能减轻不良反应,加强治疗效果。

化疗药物对骨髓间充质干细胞的损伤作用

背景:多种化疗药物在杀灭肿瘤细胞的同时会产生骨髓抑制的不良反应,不仅仅损伤造血细胞,也对骨髓间充质干细胞造成损伤,明确各种化疗药物对骨髓间充质干细胞的作用有利于合理选择、搭配化疗药物,实现高效、低毒的临床疗效。目的:探讨不同化疗药物对骨髓间充质干细胞增殖、分化和支持造血能力的影响。方法:获取正常成人骨髓间充质干细胞(n=8),用不同剂量化疗药物(环磷酰胺、马利兰、阿糖胞苷、柔红霉素、长春新碱、足叶乙甙、甲氨蝶呤、地塞米松)对骨髓间充质干细胞进行处理。检测化疗药物对骨髓间充质干细胞的增殖、凋亡作用及化疗药物撤除后骨髓间充质干细胞的恢复能力;体外诱导药物处理后骨髓间充质干细胞向脂肪细胞和骨细胞分化,通过油红O和Von Kossa染色鉴定;RT-PCR检测药物处理后骨髓间充质干细胞造血因子的表达;集落形成实验检测药物处理后骨髓间充质干细胞支持造血的功能。结果与结论:两种剂量的长春新碱、柔红霉素、足叶乙甙、阿糖胞苷和地塞米松不同程度诱导骨髓间充质干细胞凋亡,并且抑制骨髓间充质干细胞的增殖,其中长春新碱和地塞米松的抑制作用可以逆转,而柔红霉素、足叶乙甙和阿糖胞苷对间充质干细胞的抑制作用持续存在。化疗药物处理后骨髓间充质干细胞仍具有多向分化能力,可以在体外分化为脂肪细胞和成骨细胞。化疗药物处理后骨髓间充质干细胞同正常细胞一样表达造血相关因子:干细胞因子、单核细胞集落刺激因子、白细胞介素6和粒细胞集落刺激因子。柔红霉素、长春新碱、足叶乙甙和阿糖胞苷处理后骨髓间充质干细胞支持造血能力明显下降。结果显示临床常用的一些化疗药物(长春新碱、柔红霉素、足叶乙甙、阿糖胞苷)可以影响骨髓间充质干细胞的增殖和支持造血能力,而不影响骨髓间充质干细胞表达造血相关因子和多向分化能力;部分化疗药物(环磷酰胺、甲氨蝶呤和马利兰)在使用浓度下不影响骨髓间充质干细胞的上述生物学特性。