Wang S, Panter K E, Gardner D R, Evans R C, Bunch T D
USDA-ARS, US Sheep Experiment Station, Dubois, ID 83423, USA.
Anim Reprod Sci. 2004 Apr;81(3-4):237-44. doi: 10.1016/j.anireprosci.2003.10.008.
Isocupressic acid (ICA) [15-hydroxylabda-8 (17), 13E-dien-19-oic acid], a labdane diterpene acid, isolated from ponderosa pine (Pinus ponderosa), Lodgepole pine (Pinus contorta), common juniper (Juniperus communis) and Monterey cypress (Cupressus macrocarpa), induces abortion in pregnant cows when ingested primarily during the last trimester. The objective of this study was to investigate the effects of isocupressic acid on bovine oocyte maturation (in vitro maturation (IVM)-Experiment I) and preimplantation embryo development (in vitro culture (IVC)-Experiment II) using in vitro embryo production techniques and to subsequently evaluate viability and developmental competence of ICA-cultured embryos via embryo transfer to recipient heifers (Experiment III). A complete randomized block experimental design was used. In Experiment I and II, isocupressic acid was added to IVM or IVC media at 0 (TRT1, control), 1.3 (TRT2), and 2.6 microg/ml (TRT3) Results from Experiment I and II indicated that ICA did not inhibit oocyte maturation and did not adversely affect preinpiantation embryo development. Furthermore, results from Experiment II demonstrated that isocupressic acid enhanced bovine preimplantation embryo development in vitro in a dose dependent manner. Subsequently, Day 8 (Day 0 = IVF) blastocysts cultured in vitro in the medium containing 2.6 microg/ml ICA were transferred to recipient heifers and resulted in normal pregnancies as determined by ultrasound imaging. Subsequently, all but two births were normal as evaluated by post natal veterinary examination. In conclusion, ICA showed no adverse effects on oocyte maturation and preimplantation embryo development in vitro or subsequent viability in vivo using the ICA concentrations and in vitro culture parameters of this study.
异海松酸（ICA）[15 - 羟基赖百当 - 8（17），13E - 二烯 - 19 - 酸]，一种从美国黄松（Pinus ponderosa）、扭叶松（Pinus contorta）、欧洲刺柏（Juniperus communis）和大果柏木（Cupressus macrocarpa）中分离得到的赖百当二萜酸，在怀孕母牛妊娠晚期主要摄入时会导致流产。本研究的目的是使用体外胚胎生产技术研究异海松酸对牛卵母细胞成熟（体外成熟（IVM） - 实验I）和植入前胚胎发育（体外培养（IVC） - 实验II）的影响，并随后通过将ICA培养的胚胎移植到受体小母牛中来评估其活力和发育能力（实验III）。采用完全随机区组实验设计。在实验I和II中，将异海松酸以0（TRT1，对照）、1.3（TRT2）和2.6微克/毫升（TRT3）的浓度添加到IVM或IVC培养基中。实验I和II的结果表明，ICA不抑制卵母细胞成熟，也不会对植入前胚胎发育产生不利影响。此外，实验II的结果表明，异海松酸以剂量依赖的方式增强了牛体外植入前胚胎的发育。随后，将在含有2.6微克/毫升ICA的培养基中体外培养至第8天（第0天 = 体外受精）的囊胚移植到受体小母牛中，通过超声成像确定妊娠正常。随后，产后兽医检查显示，除两头外，所有出生均正常。总之，使用本研究中的ICA浓度和体外培养参数，ICA对体外卵母细胞成熟、植入前胚胎发育或随后的体内活力均无不利影响。
