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人胎盘是个独特的内分泌器官,在妊娠期完成着重要的代谢功能,包括分泌 hCC 和孕酮。本文旨在研究纯化的细胞滋养层是否具有腺苷酸环化酶和鸟苷酸调节蛋白(即 G 蛋白),进而以测定 hCG分泌水平弄清激活腺苷酸环化酶是否刺激细胞滋养层的内分泌功能。从妊娠38周自然阴道分娩或剖腹产的胎盘立即分离出细胞滋养层。经消化,离心获得高度浓缩的细胞滋养层,置于-70℃待用,或立即培养。培养中分别加入不同药品,如8-溴-cAMP(1.5mmol/1)、Forskolin(10～500μmol/l)、霍乱毒素(10～500ng/ml)、异丙肾上腺素(10～500μmol/l),百日咳毒素(100ng/ml)等。每次试验结束时,将滋养层细胞经清洗、离心后贮于-70℃。将纯化的滋养细胞用于测定腺苷酸环化酶及鉴定 G蛋白。
Human placenta is a unique endocrine organ that performs important metabolic functions during pregnancy, including the secretion of hCC and progesterone. This article aims to investigate whether purified cytotrophoblasts have adenylate cyclase and guanylin regulators (ie, G protein), and then determine whether hCG secretion levels activate adenylate cyclase to stimulate the cytotrophoblast Endocrine function. Cytotrophoblast cells were isolated immediately from a 38-week gestation of the natural vaginal delivery or caesarean section. After digestion, centrifugation to obtain highly concentrated cytotrophoblasts, placed at -70 ℃ stand-by, or immediately cultured. Different drugs such as 8-bromo-cAMP (1.5mmol / l), Forskolin (10-500μmol / l), cholera toxin (10-500ng / ml) and isoproterenol (10-500μmol / l) , Pertussis toxin (100 ng / ml) and the like. Trophoblast cells were washed at the end of each experiment and stored at -70 ° C after centrifugation. Purified trophoblasts were used to assay adenylate cyclase and identify G proteins.