肝素钠对氟尿嘧啶诱导的人脐静脉内皮细胞损伤的影响探讨

The influence of heparin sodium for fluorouracil-based induction of human umbilical vein endothelial cells damage

目的通过观察不同浓度肝素钠对氟尿嘧啶诱导的人脐静脉内皮细胞（HUVECs）损伤的影响,探讨不同浓度肝素钠稀释液及不同作用时相点对氟尿嘧啶致血管内皮细胞损伤的影响及可能的作用机制。方法将体外培养的2~3代HUVECs随机分为对照组、氟尿嘧啶组、肝素钠组,其中对照组只用培养基培养;氟尿嘧啶组用终浓度为25mg/L的氟尿嘧啶为刺激物;肝素钠组分别以终浓度为12.5U/mL、25U/mL、50U/mL肝素钠稀释液联合终浓度为25mg/L的氟尿嘧啶为刺激物。酶联免疫吸附实验（ELISA）检测细胞白介素-6（IL-6）、白介素-1（IL-1）及组织纤维溶酶原激活物（t-PA）含量。结果氟尿嘧啶组IL-6、IL-1及t-PA蛋白表达较对照组升高（P〈0.05）;与氟尿嘧啶组相比,肝素钠组可抑制氟尿嘧啶诱导的HUVECs释放IL-6、IL-1及t-PA（P〈0.05）,以50U/mL肝素钠组作用最为显著,但IL-6、IL-1及t-PA蛋白表达仍高于对照组（P〈0.05）。不同浓度肝素钠组间差异无统计学意义（P〉0.05）。同一药物浓度组,与药物作用1d后相比,第4天、第7天、第14天均有统计学差异（P〈0.05）,其中IL-6蛋白表达呈时间依赖性升高。结论肝素钠可抑制内皮细胞释放IL-1、IL-6及t-PA蛋白,对内皮细胞具有一定保护作用。

Objective To detect the mechanism on the effects of different level of heparin sodium on fluorouracil induced expression of interleukin 6 （IL-6） and interleukin 1 （IL-1）, tissue fiber dissolving enzyme activators （t- PA）, in induction of human umbilical vein endothelial cells （HUVECs）. Methods Cultured HUVECs at passage 3 to 6 was randomly divided into control group, fluorouracil group and heparin sodium group, with a final concentration of fluorouracil group is 25 mg/L of fluorouracil for stimulus, heparin sodium group on final concentration of 12.5, 25, 50 μ/ml sodium heparin dilution and the final concentration of 25 mg/L of fluorouracil for stimulus. The protein level of IL-6, IL-1 and t-PA in the cultured medium were measured by enzyme-linked immunosorbent （ELISA）. Re- suits Compared to control group, the expression of 1L-6, IL-1, t-PA protein increased in fluorouracil group （all P〈 0.05）. Compared to fluorouracil group,heparin can inhibit HUVECs induced by fluorouracil release IL-6, IL-1 and t-PA （P〈C0.05）, with 50 u/ml role for the heparin group, but IL-6, IL-1 and t-PA protein expression is still higher than that of the control group （P〈0.05）. There was no statistical difference between different concentrations of so- dium heparin group （P〈0.05）. The same drug concentration group, compared with the drug effect after 1 days, 4 days, 7 days, 14 days are statistically significant difference （P〈0.05）. IL-6 protein expression in time dependence. Conclusion Heparin group inhibits endothelial cells release, IL-6 and IL-1 t-PA protein, possesses certain protective effects on endothelial cells.