吴雨径，张晶*，赖斌，李屹，魏玉杰，刘惠亮*

    (1.武警总医院，北京100039；2.武警后勤学院，天津300309；3.湖南中医药大学第一附属医院，湖南长沙410208)

    ·基础研究·

    不同浓度的烟焦油与尼古丁对脐静脉内皮细胞增殖作用的研究

    吴雨径1，2，张晶3*，赖斌1，李屹1，魏玉杰1，刘惠亮1*

    (1.武警总医院，北京100039；2.武警后勤学院，天津300309；3.湖南中医药大学第一附属医院，湖南长沙410208)

    目的研究不同浓度烟焦油与尼古丁对人脐静脉内皮细胞增殖的影响。方法采用Ⅰ型胶原酶消化法从人脐静脉中分离人脐静脉内皮细胞，Ⅷ因子免疫细胞化学染色方法鉴定分离、培养的脐静脉内皮细胞(HUVECs)，CCK-8法、BrdU染色法检测不同浓度焦油与尼古丁对HUVECs增殖的影响。结果1、Ⅷ因子免疫细胞化学染色法显示人脐带中分离、培养的HUVECs阳性率大于95%。2、细胞培养至6 h，50 mg/L焦油组A(450)值低于对照组A(450)，具有统计学意义(P<0.05)；细胞培养至24 h，50 mg/L焦油组A(450)值低于其他各组A(450)值，具有统计学意义(P<0.05)；而不同浓度尼古丁组6 h、24 h与对照组相比随浓度的升高，A(450)值呈现逐渐升高的趋势，但无统计学意义。3、细胞培养至6 h，不同浓度焦油组BrdU阳性率无统计学意义；细胞培养至24 h，50 mg/L焦油组BrdU阳性率低于其他各组，具有统计学意义(P<0.05)；不同浓度尼古丁组6 h、24 h与对照组相比BrdU阳性率无统计学意义。结论50 mg/L焦油抑制HUVECs的增殖，尼古丁具有增强HUVECs增殖的趋势。

    焦油；尼古丁；增殖；脐静脉内皮细胞

    〔Abstract〕Objective To observe the proliferation of human umbilical vein endothelial cells(HUVECs)under different tar and nicotine conditions.M ethods HUVECs were isolated by type I collagenase digestion.After passage cultivation，we used the factorⅧrelated antibody to identify endothelial cell.CCK-8 assay and BrdU staining were used to estimate the effect of tar and nicotine on proliferation of HUVECs.Results 1.Ⅷrelated antibody staining showed that more than 95% were endothelial cells in cultured cells.2.A values at 450 nm of 50 mg/mL tar group were reduced than the control group after culturing for 6 h(P<0.05).A value of 50 mg/mL tar group were reduced compared with any other groups after culturing for 24 h(P<0.05).With the increasing of nicotine concentration ......