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

Study of Cigarette Tar and Nicotine with Different Concentrations on the Proliferation of Human Umbilical Vein Endothelial Cells

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、Brd U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Brd U阳性率无统计学意义;细胞培养至24 h,50 mg/L焦油组Brd U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Brd U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。

Objective To observe the proliferation of human umbilical vein endothelial cells （HUVECs） under different tar and nicotine conditions. Methods HUVECs were isolated by type I eollagenase digestion. After passage cultivation, we used the factor VIK related 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. Ⅷ related 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, A values were gradually increased in nicotine groups compared with control group. However, there were no significant differences. 3. BrdU positive rates of HUVECs had no significant difference between tar groups compared with control group after cultivation for 6 h. The BrdU positive rate of 50 mg/mL tar group were reduced comparing with any other groups after culturation for 24 hours （P〈0.05）. There was no significant difference of BrdU positive rate between nicotine groups and control group after cultivation for 6 h and 24 h. Conclusion The 50 mg/L Tar can inhibit the proliferation of HUVECs, however nicotine has the tendeney of stimulate the proliferation of HUVECs.