Blohberger J, Buck T, Berg D, Berg U, Kunz L, Mayerhofer A
Biomedical Center (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), Grosshaderner Strasse 9, D-82152, Planegg, Germany.
A.R.T. Bogenhausen, D-81675, Munich, Germany.
J Ovarian Res. 2016 Sep 29;9(1):62. doi: 10.1186/s13048-016-0269-0.
A previous study showed that dopamine (DA), which is contained in follicular fluid (FF) from IVF patients, strongly increased the production of reactive oxygen species (ROS) by cultured human granulosa cells (GCs). ROS, including HO, are assumed to play roles in ovarian physiology and pathology. Ovarian DA could be derived from the circulation, ovarian innervation and/or unknown ovarian sources. L-DOPA is the direct precursor of DA in its synthetic pathway. It was not yet described in FF. We examined L-DOPA levels in FF from IVF patients. As it may exert anti-oxidative and ROS-scavenging functions, we studied whether it exerts such actions in human GCs and whether DOPA-decarboxylase (DDC), the enzyme converting L-DOPA to DA, is expressed in the human ovary.
ELISA measurements revealed that human IVF-derived FF contains L-DOPA. In cultured human GCs automated confluence analyses showed that L-DOPA enhanced their survival. This is in contrast to the actions of DA, which reduced cell survival. A dose-dependent mode of action of L-DOPA was identified using a fluorescent ROS indicator. The results showed that it antagonized intracellular ROS accumulation induced by exogenous HO. DDC was absent in follicular GCs, but immunohistochemistry identified it in theca cells (TCs) of large follicles in the human ovary. Laser micro-dissection followed by RT-PCR corroborated the expression. DDC was also identified in the steroidogenic cells of the corpus luteum.
L-DOPA in FF is an antioxidant factor and exerts positive influences on GCs. Ovarian DA is derived from L-DOPA and has opposite actions. Exogenous L-DOPA is a standard therapy for Parkinson's disease, and the results raise the possibility that it may be able to exert positive actions as an antioxidant in ovarian conditions, as well.
先前的一项研究表明，体外受精（IVF）患者卵泡液（FF）中所含的多巴胺（DA）可显著增加培养的人颗粒细胞（GCs）的活性氧（ROS）生成。包括羟基自由基（HO）在内的ROS被认为在卵巢生理和病理过程中发挥作用。卵巢DA可能来源于循环系统、卵巢神经支配和/或未知的卵巢来源。左旋多巴（L-DOPA）是DA合成途径中的直接前体。此前尚未在卵泡液中发现它。我们检测了IVF患者卵泡液中的L-DOPA水平。由于它可能具有抗氧化和清除ROS的功能，我们研究了它是否在人颗粒细胞中发挥此类作用，以及将L-DOPA转化为DA的酶——多巴脱羧酶（DDC）是否在人卵巢中表达。
酶联免疫吸附测定（ELISA）显示，人IVF来源的卵泡液中含有L-DOPA。在培养的人颗粒细胞中，自动汇合分析表明L-DOPA可提高其存活率。这与DA的作用相反，DA会降低细胞存活率。使用荧光ROS指示剂确定了L-DOPA的剂量依赖性作用模式。结果表明，它可拮抗外源性HO诱导的细胞内ROS积累。卵泡颗粒细胞中不存在DDC，但免疫组织化学在人卵巢大卵泡的卵泡膜细胞（TCs）中发现了它。激光显微切割后进行逆转录聚合酶链反应（RT-PCR）证实了其表达。在黄体的类固醇生成细胞中也发现了DDC。
卵泡液中的L-DOPA是一种抗氧化因子，对颗粒细胞有积极影响。卵巢DA来源于L-DOPA，作用相反。外源性L-DOPA是帕金森病的标准治疗药物，这些结果增加了它在卵巢疾病中作为抗氧化剂也能发挥积极作用的可能性。
