激活素A和卵泡休止素对人滋养层细胞增殖和激素分泌的调节作用

Regulatory Effects of Activin A and Follistatin on the Growth and Hormone Production of Human Cytotrophoblast Cell

目的: 探讨激活素A和卵泡休止素对人早孕绒毛滋养层细胞的增殖和人绒毛膜促性腺激素及孕酮分泌的调节作用。方法: 早孕绒毛用胰蛋白酶与胶原酶联合消化后,再用小牛血清白蛋白密度梯度离心,分离纯化后所得的滋养层细胞进行体外原代培养。将不同浓度激活素A加入细胞培养液中作用48 h,观察其对滋养细胞生长和人绒毛膜促性腺激素及孕酮分泌的影响。向滋养层细胞培养液中加入激活素A和不同浓度的卵泡休止素,培养24 h,观察相互作用。结果: 激活素A和卵泡休止素都不影响滋养层细胞的增殖。激活素A以剂量依赖的方式促进滋养层细胞绒毛膜促性腺激素的分泌和孕酮的分泌,滋养细胞分别经30 ng/mL,100 ng/mL激活素A处理后,培液中绒毛膜促性腺激素和孕酮水平达到高峰(P均<0.01)。激活素A对滋养层细胞激素分泌的影响,可以被其特异结合蛋白卵泡休止素以剂量依赖方式阻断。结论: 激活素A和卵泡休止素以自分泌方式调节滋养层细胞绒毛膜促性腺激素和孕酮分泌,但并不影响细胞增殖,激活素A与卵泡休止素在人早期妊娠中起重要的生理作用。

Objective: To investigate the role of activin A and follistatin in basal hCG, progesterone productionand cell growth in first human trophblast cells. Methods: Trophoblast cells were collected for primary culture bytrypsin-collagenase digestion and bovine serum albumin (BSA) gradient centrifugation. The roles of activin A andfollistatin on trophoblast cell proliferation were determined by the MTT assay. Human chorionic gonadotropin , andprogesterone were determined by electrochemiluminescence immunoassay. For dose-response study, trophoblastcells were treated with various dose (0.1 to 100 ng/mL) of activin A for 48 hours. To determine the interactionbetween activin A and follistatin, cells were treated with activin A (10 ng/mL) alone, or activin A with differentconcentrations of follistatin (10, 20, 50 ng/mL) for 24 hours. Results: Neither activin A nor follistatin affectedhuman trophoblast cell growth. Human chorionic gonadotropin and progesterone levels in cultured media increasedin a dose-dependent manner by activin A with the maximal effect observed at the dose of 30 ng/mL, 100 ng/mL. Theeffects of activin A on hCG and progesterone productions were blocked by follistatin, an activin A binding protein,in a dose-dependent manner. Conclusion: As autocrine factors, activin A and follistatin are involved in the regulationof hCG and progesterone productions in human trophoblast cells but they do not have influence on cell proliferation.The finding provides evidence that activin A and follistatin may play important role in early pregnancy.