丹参酮ⅡA对经子痫前期患者血清刺激的人脐静脉内皮细胞的影响

Impact of Tanshinone ⅡA on Human Umbilical Vein Endothe lial Cells Injured by Serum of Preeclampsia

目的观察丹参酮(Tan)ⅡA对经子痫前期患者血清刺激的人脐静脉内皮细胞(HUVEC)的影响。方法选取2010年12月至2011年4月于哈尔滨医科大学附属第一医院住院分娩的20例子痫前期患者的60份血清标本(每例患者采集3份)为研究对象。根据是否采用TanⅡA对HUVEC进行预处理,将其分为子痫前期组(n=20)、1.0mg/LTanⅡA处理组(n=20)和2.0mg/LTanⅡA处理组(n=20)。选取同期在相同医院就诊的20例正常晚孕期妇女的血清标本(每例孕妇采集1份)作为空白对照组(n=20)(本研究遵循的程序符合本院人体试验委员会所制定的伦理学标准,得到该委员会批准,分组征得受试对象本人的知情同意,并与之签署临床研究知情同意书)。20例子痫前期患者与20例正常晚孕期妇女的年龄、孕龄等比较,差异无统计学意义(P>0.05)。倒置显微镜下观察4组HUVEC形态。采用CCK-8法测定4组HUVEC光密度(OD)值(HUVEC活力),并对结果进行统计学分析。结果倒置显微镜下见:子痫前期组HUVEC呈损伤改变,细胞稀疏,细胞质与细胞核界限模糊,细胞质中有暗色颗粒;1.0mg/LTanⅡA处理组和2.0mg/LTanⅡA处理组HUVEC细胞形态呈多角形、椭圆形,排列紧密,细胞边界清楚。对4组血清标本继续培养24h和48h所测OD值,子痫前期组与空白对照组比较,差异有统计学意义(P<0.05);1.0mg/LTanⅡA处理组、2.0mg/LTanⅡA处理组分别与子痫前期组比较,差异均有统计学意义(P<0.05);1.0mg/LTanⅡA处理组与2.0mg/LTanⅡA处理组比较,差异亦有统计学意义(P<0.05)。结论子痫前期患者血管内皮细胞受损,导致内皮细胞活力下降,而TanⅡA可减轻该损伤作用。

Objective To explore the impact of tanshinone Ⅱ A（Tan ⅡA）on the proliferation of human umbilical vein endothelial cells（HUVEC） effected by serum of preeclampsia.Methods From December 2010 t o March 2011,a total of 60 serum samples which took from 20 patients with preec l ampsia（3 serum samples each patient） were divided into preeclampsia group（n= 20）,1.0 mg/L TanⅡA group（n=20） and 2.0 mg/L TanⅡA group（n=20） accord in g to the usage of TanⅡA.Meanwhile,20 serum samples taken from 20 normal late pregnancy women were chose in the same hospital for treatment（1 serum sam ple each patient） as blank control group.The study protocol was approved by the Ethical Review Board of Investigation in Human Being of First Hospital of Ha rbin Medical University.Informed consent was obtained from each patients.There was no significant difference in age and gestational age between preeclampsia p ati ents and normal late pregnancy women（P0.05）.The morphologic changes of HU VEC were observed under inverted microscope.Cell counting kit-8（CCK-8） assay was applied to detect HUVEC optical density（OD） value（HUVEC vitality） i n each group,and analyzed that statistically.Results Under in verted microscope,cells of preeclampsia group was injured,showed that cell was sparse,cytoplasm and nucleus became blurred,and dark particles was found in cy toplasm.For 1.0 mg/L TanⅡA group and 2.0 mg/L TanⅡA group,The morphologi c ch anges of HUVEC were polygonal,oval,closely arranged,and clear cell boundaries under the microscope.OD value in four serum samples were cultured at 24 h and 48 h,there was significant difference between preeclampsia group an d blank control group（P0.05）;OD value in both 1.0 mg/L TanⅡA treatme nt g roup and 2.0 mg/L TanⅡA group were obviously higher than those in pre eclampsia group,respectively（P0.05）.There also had significant diffe renc e between 1.0 mg/L TanⅡA treatment group with 2.0 mg/L TanⅡA treatment group（P0.05）.Conclusions The stimulation of preeclampsia serum might cause the decrease viability of endothelial HUVEC.Tan ⅡA can protect HUVEC from dam age induced by preeclampsia serum.