脐静脉内皮细胞及其培养液中血栓素测定的初步探讨

MEASUREMENT OF THROMBOXANE IN MEDIUM HUMEN UMBILICAL VEIN ENDOTHELIAL CELLS AND ITS CULTURE MEDIUM

目的研究人脐静脉内皮细胞（ＨＵＶＥＣ）能否分泌血栓素及妊高征患者血清对其分泌活动的影响。方法体外原代培养５例妊高征患者ＨＵＶＥＣ和５例正常妊娠ＨＵＶＥＣ，每例分为３组，当其融合时，分别与妊高征患者（第１组）和正常孕妇（第２组）的产前血清以及培养液（第３组）共同孵育２４ｈ。采用放免法测定培养液中血栓素含量，以第３组为基础分泌量。并用免疫荧光染色鉴定ＨＵＶＥＣ内的血栓素颗粒。结果ＨＵＶＥＣ胞质内有大量血栓素颗粒；妊高征ＨＵＶＥＣ和正常妊娠ＨＵＶＥＣ的ＴＸＢ２基础分泌量无统计学差异。与妊高征ＨＵＶＥＣ的基础分泌量相比，在妊高征ＨＵＶＥＣ的培养液中加入妊高征患者产前血清，血栓素分泌量无变化［（２１６．０３±７８．９６）ｐｇ／０．１ｍｌ，Ｐ＞０．０５］，而正常孕妇产前血清可使其明显下降［（１９１．３７±２５．８０）ｐｇ／０．１ｍｌ，Ｐ＜０．０５］；与正常妊娠ＨＵＶＥＣ的基础分泌量相比，在正常ＨＵＶＥＣ的培养液中加入妊高征患者产前血清，ＴＸＢ２产量下降为（２４３．２５±２４．１１）ｐｇ／０．１ｍｌ，Ｐ＜０．０５，而加入正常妊娠产前血清，正常妊娠ＨＵＶＥＣ的ＴＸＢ２量降低为（２１８．３±２０．０８）ｐｇ／０．１ｍｌ，Ｐ＜?

URPOSE To study whether or not the umbilical vein endothelial cell (HUVEC) can secrete thromboxane and the effect of sera from patients with pregnancy-induced hypertension (PIH) on its secretion.METHODS HUVEC were culture from 5 patients with PIH and 5 normal pregnancies. When the cells are confluent, Sera of PIH and normal pregnancy were added respectively, after 24 hours incubation, the medium were taken to measure the thromboxane products by radioimmunoassay. Besides, immunohistochemical method has been used to judge the thromboxane granule in HUVEC.RESULTS There are a lot of thromboxane granule in the cytoplasma of HUVEC. The basal production of thromboxane in PIH and normal pregnancy is (325.03±63.92)pg/0.1ml and (339.2±4.24)pg/0.1ml respectively, P>0.05. Comparing to the basal production of thromboxane in PIH, when the sera of PIH added, the thromboxane production in medium of HUVEC from PIH has no change [(216.03±78.96)pg/0.1ml], but when the sera of normal pregnancy added, it decreased obviously [(191.37±25.80)pg/0.1ml], P<0.05; Comparing to the basal production of thromboxane in normal pregnancy, when the sera of PIH added, the thromboxane production in medium of HUVEC from normal pregnancy decreased to (243.25±24.11)pg/0.1ml, P<0.05; when the sera of normal pregnancy added, it cut down to (218.3±20.08)pg/0.1ml, P<0.01.CONCLUSIONS This is the first report on that HUVEC may secrete thromboxance, moreover, there are probably some cytotoxic factors existed in the sera of PIH, affecting thromboxance secretion, therefore play an important role in the etiology of PIH.