再生障碍性贫血患者骨髓CD106+间充质干细胞对其骨髓血管衰竭的作用  

Effect of CD106^+ Mesenchymal Stem Cell on Bone Marrow Vascular Failure in Patients with Aplastic Anemia

作　　者：卢士红 葛美丽 郑以州 杨少光 陈芳 尤亚红 韩忠朝 LU Shihong;GE Meili;ZHENG Yizhou;YANG Shaoguang;CHEN Fang;YOU Yahong;HAN Zhongchao(State Key Laboratory of Experimental Hematology,Institute of Hematology and Blood Diseases Hospital, CAMS and PUMC,Tianjin 300020,China)

机构地区：[1]中国医学科学院,北京协和医学院,血液病医院,血液学研究所国家重点实验室,天津300020

出　　处：《中国医学科学院学报》2018年第2期178-186,共9页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(2013-81330015,2014-81300388)

摘　　要：目的探讨再生障碍性贫血(AA)患者骨髓间充质干细胞(MSCs)形成血管的能力及受影响的因素。方法扩增培养AA患者骨髓与正常人骨髓MSCs,流式及免疫组织化学鉴定检测CD106基因表达情况及CD106^+MSCs含量;测定新鲜AA骨髓中CD105^+CD106^+MSCs含量,检测AA MSCs和正常人MSCs被诱导成内皮细胞能力,检测封闭CD106抗原的正常人MSCs被诱导成内皮细胞能力及在基质胶中形成管状结构的能力。结果 AA骨髓MSCs CD106基因表达较正常人缺失(降低12.13倍),CD106^+MSCs含量降低[(28.03±17.71)%比(59.61±12.26)%,P=0.000],新鲜骨髓中CD105^+CD106^+MSCs含量降低[(0.33±0.10)%比(2.98±0.46)%,P=0.0005]。AA MSCs被诱导成CD31+内皮细胞能力明显迟缓[(13.67±1.50)%比(43.24±0.96)%,P=0.0004],而将正常人MSCs封闭CD106抗原后其被诱导成CD31+细胞能力[(26.00±2.65)%比(91.78±2.44)%,P=0.000]及管状结构形成能力[(13.81±1.98)mm比(68.12±6.78)mm,P=0.0015]也均显著下降。结论 AA患者骨髓CD106基因表达缺失、CD106^+MSCs含量降低可促进患者骨髓血管衰竭。Objective To investigate the vascularization ability of mesenchymal stem cells(MSCs)and explore its influencing factors in aplastic anemia(AA)patients.Methods MSCs were isolated from the bone marrow of AA patients(AA MSCs)and normal controls(N MSCs)were cultured and then evaluated by flow cytometry and immunofluorescene staining technique.The expression level of vascular cell adhesion molecule-1(CD106)was detected by gene sequencing,and the content and fluorescene intensity of CD106+MSCs was determined by fluorescence-activated cell sorting.The content of CD105+CD106+MSCs in fresh AA bone marrow was measured,followed by the determination of the capability of endothelial differentiation from AA MSCs and N MSCs with immunofluorescene analysis;finally,the capability of CD31+cell differentiation from CD106-blocking N MSCs and its tubular structures formation in matrigel were tested.Results The expression of CD106 in AA patients was defective(decreased by 12.13 times when compared with N MSCs)and the concentration and fluorescene degree of CD106+MSCs was also decreased in AA patients[(28.03±17.71)%vs.(59.61±12.26)%,P=0.000].The content of CD105+CD106+MSCs decreased significantly in the fresh bone marrow[(0.33±0.10)%vs.(2.98±0.46)%,P=0.0005].Besides,the capability of CD31+cell differentiation from AA MSCs was significantly delayed[(13.67±1.50)%vs.(43.24±0.96)%,P=0.0004].Also,the capability of CD31+cell differentiation and tubular structures formation of CD106-blocking N MSCs was also obviously decreased[(26.00±2.65)%vs.(91.78±2.44)%,P=0.000;(13.81±1.98)mm vs.(68.12±6.78)mm,P=0.0015].Conclusion The deficient or decreased expression of CD106+MSCs accelerate the bone marrow vascularization failure in AA patients.

关 键 词：骨髓间充质干细胞 血管细胞黏附分子-1 管状结构 血管生成 

分 类 号：R552[医药卫生—血液循环系统疾病] R556[医药卫生—内科学]