论文部分内容阅读
背景:有研究指出氯吡格雷、噻氯匹定可引起粒细胞减少,阿司匹林可抑制内皮祖细胞增殖。目前缺乏这些抗血小板药物会对移植入心肌的骨髓间充质干细胞产生何种作用的报道。目的:探讨氯吡格雷、噻氯匹定和阿司匹林对人骨髓间充质干细胞增殖及分泌功能的影响。设计、时间和地点:细胞学体外观察,于2006-03/12在上海市疾病预防控制中心毒理学实验室完成。材料:第2代人骨髓间充质干细胞由上海第九人民医院组织工程研究中心提供。氯吡格雷、噻氯匹定为杭州赛诺菲圣德拉堡民生制药有限公司产品,批号分别为国药准字J20040006和国药准字H19980186。阿司匹林由拜耳医药有限公司生产,批号为国药准字20050059。方法:骨髓间充质干细胞解冻后,加入含100U/mL青霉素、100mg/L链霉素、10%胎牛血清的低糖DMEM体外扩增培养,传至第6代,分别用不同浓度的氯吡格雷、噻氯匹定、阿司匹林进行干预处理,并设立空白对照。主要观察指标:MTT比色法检测细胞增殖情况,流式细胞仪检测细胞表面抗原表达,ELISA法测定细胞培养液中血管内皮生长因子浓度。结果:反映骨髓间充质干细胞增殖变化的A570值在0.02,0.1,0.4,2,10,40μmol/L氯吡格雷或噻氯匹定作用后均明显高于空白对照(P<0.01);经60,600,2000μmol/L阿司匹林处理后则低于空白对照(P<0.05)。细胞表面抗原CD34、CD105、CD106阳性细胞百分率经3种抗血小板药物作用后与空白对照基本相似。与空白对照比较,经0.02μmol/L氯吡格雷及5,2000μmol/L阿司匹林处理后血管内皮生长因子含量无明显变化(P>0.05);40μmol/L氯吡格雷或噻氯匹定作用后血管内皮生长因子含量均显著降低;0.02μmol/L噻氯匹定作用后血管内皮生长因子含量显著升高(P<0.01)。结论:氯吡格雷和噻氯匹定对人骨髓间充质干细胞具有明显的促增殖作用,阿司匹林可抑制其增殖。高浓度氯吡格雷和噻氯匹定能够抑制人骨髓间充质干细胞分泌血管内皮生长因子,低浓度噻氯匹定可促进其分泌。此3种抗血小板药物对人骨髓间充质干细胞的进一步分化无明显影响。
Background: Some studies have pointed out that clopidogrel and ticlopidine can cause neutropenia, and aspirin can inhibit the proliferation of endothelial progenitor cells. The current lack of these anti-platelet drugs will be transplanted into the heart of bone marrow mesenchymal stem cells what kind of effect has been reported. Objective: To investigate the effects of clopidogrel, ticlopidine and aspirin on the proliferation and secretion of human bone marrow mesenchymal stem cells. DESIGN, TIME AND SETTING: The cytology in vitro observation was performed at the Toxicology Laboratory of Shanghai Center for Disease Control and Prevention from March to December 2006. Materials: The second generation of human bone marrow mesenchymal stem cells provided by the Shanghai Ninth People’s Hospital Tissue Engineering Research Center. Clopidogrel, ticlopidine Hangzhou Sanofi Sandela Fort Minsheng Pharmaceutical Co., Ltd. products, batch numbers were Zhunzi J20040006 and Zhunzi H19980186. Aspirin Bayer Pharmaceutical Co., Ltd. production, batch number for the country Yaozhun 20050059. Methods: Bone marrow mesenchymal stem cells were thawed and cultured in low glucose DMEM containing 100 U / mL penicillin, 100 mg / L streptomycin and 10% fetal bovine serum for 6 generations. The cells were treated with different concentrations of clopidogrel Gray, ticlopidine, aspirin intervention, and the establishment of a blank control. MAIN OUTCOME MEASURES: Cell proliferation was detected by MTT colorimetric assay, cell surface antigen expression was detected by flow cytometry, and vascular endothelial growth factor (VEGF) concentration was measured by ELISA. Results: A570, which reflects the proliferation and proliferation of BMSCs, was significantly higher than that of blank control (P <0.01) after treated with 0.02, 0.1, 0.4, 2, 10 and 40 micromol / L clopidogrel or ticlopidine 60,600,2000μmol / L aspirin treatment was lower than the blank control (P <0.05). Cell surface antigen CD34, CD105, CD106 positive cells by the three anti-platelet drugs after the role of the blank control is basically similar. Compared with the blank control group, there was no significant change in the content of vascular endothelial growth factor after treated with 0.02μmol / L clopidogrel and 5,2000μmol / L aspirin (P> 0.05). After 40μmol / L clopidogrel or ticlopidine treatment, The content of VEGF was significantly decreased. The content of VEGF was significantly increased after 0.02μmol / L ticlopidine treatment (P <0.01). CONCLUSION: Clopidogrel and ticlopidine have a significant proliferative effect on human bone marrow mesenchymal stem cells, and aspirin can inhibit its proliferation. High concentrations of clopidogrel and ticlopidine can inhibit human bone marrow mesenchymal stem cells secrete vascular endothelial growth factor, low concentrations of ticlopidine can promote its secretion. The three kinds of anti-platelet drugs on human bone marrow mesenchymal stem cells further differentiation had no significant effect.