医药学论文：VEGF在妊娠肝内胆汁淤积症中的作用以及丹参治疗机制的研究

【摘要】 目的 探讨VEGF在妊娠肝内胆汁淤积症中的作用以及丹参活血化瘀改善胎儿血供的可能机制。方法 采集妊娠肝内胆汁淤积症患者血清，Jaffe法分离培养脐静脉内皮细胞，用ICP患者血清刺激培养的内皮细胞，模拟ICP的体内环境，行丹参干预，与正常妊娠血清组比较，不同含量作用不同的时间，MTT法检测细胞活力，ELISA法检测分泌型VEGF蛋白表达。结果 ICP患者血清与HUVECs共培养：（1）MTT法发现随着血清含量的增加A值逐渐降低（0～20%），且低于相应浓度的正常妊娠血清组，有统计学差异；（2）ELISA法发现：同正常孕妇血清组相比，ICP患者血清组HUVECs表达分泌型VEGF蛋白降低，且随着血清含量的增加和培养时间的延长，VEGF表达逐渐降低。不同浓度的丹参干预后随着含量的增加（1～8mg/ml），细胞活力和VEGF蛋白表达逐渐升高，以8mg/ml组作用最佳。结论 ICP患者血清对HUVECs具有损伤作用，使细胞活力降低。其损伤作用可能与VEGF的分泌减少有关。丹参对内皮细胞具有保护作用并能增加 VEGF蛋白的表达。丹参可能通过增加VEGF蛋白的表达具有活血化瘀作用，改善胎盘和胎儿的血供而改善围生儿预后。

 

　　【关键词】 妊娠肝内胆汁淤积症；脐静脉；血管内皮细胞；血管内皮生长因子；丹参
Study of the effects of vascular endothelial growth factor and the therapeutic mechanisms of Salvia Miltiorrhiza Bge in intrahepatic cholestasis of pregnancy

　　【Abstract】 Objective To sudy the effects of vascular endothelial growth factor and the therapeutic mechanisms of Salvia Miltiorrhiza Bge in intrahepatic cholestasis of pregnancy(ICP). Methods Twelve patients with ICP and twelve normal pregnancy women take part in this study,their serum is collected before therapy when in hospital. Endothelial cells are isolated from human umbilical veins choosed among the 10 normal pregnancy according to Jaffe's method, serum of different concentration of ICP groups and normal pregnancy groups is cocultured with HUVECs for different time point respectively. MTT method is applied to measuring the vitality of HUVECs.The expression value of vascular endothelial growth factor(VEGF) is detected by enzyme-linked immunosorbent assay (ELISA) . Results Compared with normal pregnancy , the value of MTT in ICP serum groups,the A value is significantly lower,P< 0.05. The value of soluble VEGF in ICP serum group is lower than that in the nomal pregnancy serum group.With the serum concetration increases, the value of sVEGF decreases in the ICP groups . Compared with ICP group, the value of MTT and sVEGF in the Salvia Miltiorrhiza Bge therapeutic group increases with the concentration increase,8mg/ml of Salvia Miltiorrhiza Bge is the optical concentration. Conclusion Serum of ICP patients impairs HUVECs in vitro and decreases the expression of VEGF. The mechanism of serum of ICP patients impairs HUVECs maybe by decreasing the expression of VEGF. Salvia Miltiorrhiza Bge could protect HUVECs against the injury of ICP serum and promotes the expression of VEGF. Salvia Miltiorrhiza Bge improves HUVECs by increasing the value of VEGF.

 

　　【Key words】 intrahepatic cholestasis of pregnancy; umbilical vein; vascular endothelial cell(VEC);vascular endothelial growth factor; Salvia Miltiorrhiza Bge

 

　　妊娠肝内胆汁淤积症（intrahepatic cholestasis of pregnancy，ICP）是一种妊娠特异性的疾病［1］，以孕妇出现全身或局部皮肤瘙痒、黄疸及胆酸增高为特征。ICP可引起严重的围生儿并发症，围生儿发病率和死亡率明显增加。关于ICP胎儿宫内缺氧的机制尚不清楚。胎盘-胎儿血供受到一系列生长因子的调节，血管内皮生长因子（vascular endothelial growth factor， VEGF）是一种直接而特异作用于血管内皮细胞的高效促有丝分裂因子，在刺激新血管发生与生长及维持血管壁的完整性和通透性方面均有重要意义。VEGF对于受精、着床、胎盘血管形成及胎儿生长发育起着重要的作用。表达异常则可能导致胎儿宫内氧供不良，对围生儿产生不良影响。具有行气活血作用的中药-丹参，临床上被用于治疗ICP以改善围生儿预后。本文以ICP患者血清与正常脐静脉内皮细胞共培养并用不同含量的丹参注射液进行干预，探讨VEGF在妊娠肝内胆汁淤积症中的作用，以及丹参活血化瘀改善胎儿血供的可能机制。