【作者】 杜雪寒；

【导师】 蒋犁；

【作者基本信息】 东南大学 ， 儿科学， 2004， 硕士

【摘要】 目的:探讨ICP对围生儿的不良影响以及胎儿宫内缺氧的可能机制,探讨丹参活血化淤改善胎儿血供的可能机制。方法:临床收集ICP和正常妊娠各30例,采集外周静脉血,分离血清;比较新生儿体重、身长,胎盘重量,羊水粪染和Apgar评分情况。随机选取正常妊娠脐带,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,并与正常妊娠血清组比较,不同浓度作用不同的时间,MTT法检测细胞活力,ELISA法和免疫细胞化学法检测分泌型和膜结合型VEGF蛋白表达。结果:①ICP患者娩出的新生儿体重和胎盘重量明显低于正常妊娠组,羊水粪染率明显高于正常组,有统计学差异。②ICP患者血清与HUVECs共培养24h,MTT法发现浓度在(0、5、10、15、20%)范围内,随着浓度的增加OD值逐渐降低,以 20%浓度组OD值为最低;且OD值低于相应浓度的正常血清组,有统计学差异。③通过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;分别刺激 12h、24h、36h,20%浓度组的VEGF蛋白表达均低于10%浓度组;24h、36h的VEGF蛋白量均低于相应浓度的12h组;20%ICP血清刺激24h,VEGF可降至最低。④与ICP血清组相比,丹参干预组随着浓度由1mg/ml增加到8mg/ml,吸光度值和VEGF蛋白表达逐渐升高,以8mg/ml组作用最佳,与正常孕妇血清组无差别。⑤应用免疫细胞化学方法:ICP患者血清刺激后HUVECs呈淡棕黄色着色,弱阳性,着色明显比正常孕妇血清组浅,丹参干预后,细胞着色变深,以8mg/ml组着色最深,呈强阳性,与正常孕妇血清刺激组HUVECs着色无统计学差异。结论:①ICP对围生儿产生不良影响。②ICP患者血清对HUVECs具有损伤作用,使细胞活力降低。丹参对内皮细胞具有保护作用。③ICP患者血清对内皮细胞的损伤,可能与VEGF的分泌减少有关。丹参能增加HUVECs VEGF蛋白的表达。④丹参可能通过增加VEGF蛋白的表达,具有活血化淤作用,改善胎盘和胎儿的血供而改善围生儿预后。更多还原

【Abstract】 University: Southeast universityObjective:To explore poor perinatal outcome in ICP and it′s mechanisms responsible for fetal intrauterine asphyxia;To study the mechanism of Salvia Miltiorrhiza Bge(丹参)in improving fetal blood microcirculation in ICP.Methods: Thirty patients with ICP and thirty normal pregnancy women take part in this study.Their periblood is collected and centrifugated.Sixty newborns’ weight,stature,Apgar score,placental weight and meconium staining of amniotic fluid are compared.Endothelial cells are isolated from human umbilical veins 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.Salvia Miltiorrhiza Bge of 1,2,4,8mg/ml is used to intervene HUVECs in ICP group.MTT method is applied to measuring the vitality of HUVECs.The expression value of vascular endothelial growth factor(VEGF)of different groups is detected by immunocytochemistry and Enzyme-linked immunosorbent assay (ELISA) respectively.Results: ①Compared with normal pregnancy,neonatal weight and placental weight are markedly lower,the incidence rate of meconium staining of amniotic fluid is distinctly higher in ICP group.②The value of MTT in ICP serum groups is significantly lower than that in the normal pregnancy serum group.With the increase of serum concentration,the value of MTT decreases.③Compared with the normal group,the value of sVEGF in ICP serum group is lower.Furtheremore,the value of sVEGF decreases at most if HUVECS were cocultured with 20%ICP serum for 24 hours.④In the Salvia Miltiorrhiza Bge therapeutic groups,the value of MTT and VEGF increases with the concentration increase,8mg/ml of alvia Miltiorrhiza Bge is the optical concentration and being no significantly different from the normal serum group.⑤By immunocytochemistry,the expression of VEGF in HUVECs decreases in the ICP serum group,8mg/ml of Salvia Miltiorrhiza Bge enhances the expression of VEGF at most. Conclusion: ①ICP had bad effects on the fetus.②Serum of ICP patients impairs HUVECs in vitro, but Salvia Miltiorrhiza Bge could protect HUVECs against the injury of ICP serum.③The serum of ICP patients decreases the expression of VEGF in HUVECs.While Salvia Miltiorrhiza Bge promotes the expression of VEGF.④Salvia Miltiorrhiza Bge improves fetal blood microcirculation in ICP group by increasing the value of VEGF .更多还原