巨噬细胞对内皮细胞部分生物学特性的影响  

Influence of macrophages on some biological features of endothelial cells

作　　者：刘亮[1] 王颖[1] 张晓启[2] 刘旭盛[3] 

机构地区：[1]南方医科大学南方医院烧伤科,广州510515 [2]沈阳武警辽宁总队医院烧伤科 [3]广州中山大学第一附属医院烧伤科

出　　处：《中华烧伤杂志》2008年第1期48-50,共3页Chinese Journal of Burns

摘　　要：目的建立人巨噬细胞系（U937）与人脐静脉血管内皮细胞系（ECV304）体外共培养模型，以伴刀豆球蛋白A（ConA）作为U937细胞激活剂，观察其调节血管生成的可行性。方法培养ECV304细胞至60％融合，分别加入ConA25μg／mL、U937细胞（1×10^5个）、U937细胞（1×10^5个）＋ConA25μg／mL，共同培养48h。以常规培养的ECV304细胞为对照组。用流式细胞仪检测细胞周期及细胞凋亡的变化；采用氚标记胸腺嘧啶脱氧核苷掺入法检测内皮细胞DNA合成情况；用反转录一聚合酶链反应技术检测内皮细胞同源盒HOXB2基因mRNA的表达。结果ConA＋U937细胞可使ECV304细胞S期百分比升高至（48．860±2．290）％，与对照组（41．590±2．590）％比较，差异有统计学意义（P〈0．01）；DNA合成亦明显增加[放射性荧光闪烁计数值为（5694±917）min^-1]，与对照组[（2498±1109）min^-1]比较，差异有统计学意义（P〈0．01）；内皮细胞HOXB2基因mRNA的表达水平为0．947±0．003，与对照组0．646±0．004比较，差异有统计学意义（P〈0．01）。各组细胞凋亡率相近（P〉0．05）结论经ConA活化的U937细胞可促进ECV304细胞增殖进而调节血管生成，HOXB2与内皮细胞增殖有密切关系。Objective To establish the co-culture model of human macrophage cell line （U937） with human vein umbilical cell line （ECV304） , and to explore the feasibility of using concanavalin A （ConA） as U937 cell stimulator in regulating angiogenesis. Methods ECV304 cells were cultured in vitro ,and to which were respectively added U937 cells（ 1 ± 105/） , 25μg/mL ConA , and U937 cell（ 1 ± 105 ） ± ConA （25μg/mL） after cell fusion rate reaching 60% , and then co-cultured for 48 hours. ECV 304 cells in conventional culture were used as controls. 3 H-TdR incorporation test was employed to determine the DNA synthesis of vascular endothelial cells. Flow cytometry was used to determine the changes in the cell cycle, and RT-PCR was adopted to determine the expression of homebox （HOXB2） mRNA. Results After conA stimulation to ECV 304 co-cultured with U937 cells , the percentage of cells in S phase （48. 860 ± 2. 290） , the DNA synthesis[ （5694 ±917）min^-1 ] , and the expression of HOXB2 mRNA （0.947 ±0.003） were obviously higher than those in control group[ 41. 590 ± 2. 590 vs （2498 ± 1109） min ^-1 vs 0. 646 ± 0. 004, P 〉 0.01 ]. There was no obvious difference in apoptosis among above stimulation methods （ P 〉 0. 05 ） Conclusion U937 cells activated by ConA can promote the proliferation of ECV304 cells and further regulate angiogenesis. HOXB2 gene is closely related to the endothelial proliferation.

关 键 词：巨噬细胞 内皮细胞 共同培养技术 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]