摘要
目的探讨盐酸贝那普利(洛汀新,血管紧张素转换酶抑制剂)对不同系别造血祖细胞分化的影响及其分子机制。方法由正常脐血分离出单个核细胞,在含有不同浓度盐酸贝那普利与其他因子包括干细胞因子(SCF)、白细胞介素-3(IL-3)、粒-巨噬细胞集落刺激因子(GM-CSF)、红细胞生成素(EPO)、血管紧张素Ⅱ(AngⅡ)的培养基中悬浮培养,然后转到半固体培养基,对粒-巨噬细胞集落形成单位(CFU-GM)和红细胞样爆发形成单位(BFU-E)集落进行观察并计数,同时应用流式细胞仪检测红系和粒系所占比例;巢式反转录聚合酶链反应(RT-PCR)监测AngⅡ1型受体(AT1R)mRNA表达。结果①红系祖细胞在悬浮培养6、9d时均可检测到AT1R的表达,两时间点各组AT1R的表达量差异均无统计学意义;在上述两时间点各对应组之间AT1R的表达量差异无统计学意义;②半固体培养14d后,各组的粒系集落明显减少,而各组之间比较差异无统计学意义;③各组红系集落形成丰富,但各组间差异无统计学意义;④各组红系和粒系所占的百分比差异有统计学意义(85.85%vs13.15%,85.78%vs13.21%,85.77%vs13.20%,85.79%vs13.18%,85.81%vs13.23%,P<0.05)。结论在体外细胞培养过程中AngⅡ能促进红系祖细胞的扩增,而盐酸贝那普利不能阻断AngⅡ对红系造血祖细胞的优势分化作用。
Objective To investigate the effect and molecular mechanism of lotensin （benazepfil,an angiotensin converting enzyme inhibitor, ACEI） on the expansion of hematopoietic progenitor cells in vitro. Method Mononuclear cells from normal umbilical cord blood were suspansion cultured in medium containing lotensin of different concentrations and stimulating factors（IL-3,SCF,GM-CSF,EPO,Ang Ⅱ ）. Then, cells were transferred to methylcellulose semisolid medium and cultured for fourteen days. Floweytometry was used to detect the proportion of crythroid and myeloid. The numbers of colony-forming unit-granulocyte-macrophage（CFU-GM） and burst-forming uniterythroid（BFU-E） colonies were counted. Nested RT-PCR was used to determine the expression of angiotensin Ⅱ type 1 receptor （ATIR）. Results （1） AT1R mRNA was detectable at day 6 and 9. At each time point. The expressions of AT1R showed no significant difference;.At both time point, AT1R expressions exhibited no marked difference; （2）When ceils were transferred to methylcellulose semisolid medium and cultured for 14 days later, the numbers of granulocyte decreased significantly in every group, there is no significant difference among them; At the same time, erythroid were marked increased, but the differences among them were not remarkable;（3）The proportion of granulocyte and erythmid were markedly different among the groups（85.85% vs 13.15%, 85.78% vs 13.21%, 85.77% vs 13.20%, 85.79%vs 13.18%, 85.81% vs13.23%,P〈0.05）. Conclusion Angiotensin II had stimulatory effect on erythroid progenitors in cultured ceils, while Lotensin has no effect on it.
出处
《中国药物与临床》
CAS
2006年第1期30-33,共4页
Chinese Remedies & Clinics
基金
山西省自然科学基金资助项目(20051101)
