Kataria Hardeep, Gupta Muskan, Lakhman Sukhwinder, Kaur Gurcharan
Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab 143005, India.
Department of Pharmaceutical, Social and Administrative Sciences, D'Youville College, School of Pharmacy, Buffalo, NY 14201, USA.
Neurochem Int. 2015 Oct;89:111-9. doi: 10.1016/j.neuint.2015.08.001. Epub 2015 Aug 6.
Ashwagandha (Withania somnifera) has a long history in traditional medicines as an aphrodisiac. It has been known to influence sexual behaviour in animal models but mechanism of action is still unknown. The present study was aimed to investigate the mechanisms by which Ashwagandha extract exert its gonadotropic activities. Due to the complexity of neuroendocrine pathways, there are limited in vitro models available despite the strong demand for such systems to study and predict neuroendocrine effects of chemicals or natural products. Immortalized rat hypothalamic GnV-3 cell line was investigated as a model to screen for neuroendocrine effects of Ashwagandha extract. GnV-3 cells were cultured under different media conditions and evaluated after treatment with Ashwagandha water extract, for GnRH expression and release by immunostaining and ELISA respectively. These cells acquired differentiated morphology, characteristic shape displayed by preoptic GnRH neurons in vivo. In addition, GnV-3 cells exhibited upregulation of plasticity related polysialylated neural cell adhesion molecule (PSA-NCAM) and mature dendrite marker microtubule associated protein (MAP2) as well as GnRH expression and release. Chloroform fraction of the extract proved to exhibit all the bioactive properties as it induced differentiation and upregulated GnRH and MAP2 expression in GnV-3 cells, similar to Ashwagandha extract. Withanone and Withaferin A were found to be present in ASH-WEX and chloroform fraction while Withanone came out to be the major constituent of chloroform fraction. The preliminary in vivo studies in adult male animals showed that ASH-WEX was able to upregulate the GnRH levels although non-significantly. Taken together, this data demonstrate significant morphological and physiological changes in GnV-3 cells after treatment with Ashwagandha extract and may suggest the potential beneficial effects of Ashwagandha on reproductive functions in vivo.
南非醉茄（Withania somnifera）作为一种壮阳药在传统医学中有着悠久的历史。已知它会影响动物模型的性行为，但其作用机制仍不清楚。本研究旨在探究南非醉茄提取物发挥促性腺活性的机制。由于神经内分泌途径的复杂性，尽管对研究和预测化学物质或天然产物的神经内分泌效应的此类系统有强烈需求，但可用的体外模型有限。研究了永生化大鼠下丘脑GnV - 3细胞系作为筛选南非醉茄提取物神经内分泌效应的模型。GnV - 3细胞在不同培养基条件下培养，并在用南非醉茄水提取物处理后分别通过免疫染色和酶联免疫吸附测定（ELISA）评估促性腺激素释放激素（GnRH）的表达和释放。这些细胞呈现出分化的形态，即体内视前区GnRH神经元所显示的特征形状。此外，GnV - 3细胞表现出可塑性相关的多唾液酸神经细胞黏附分子（PSA - NCAM）和成熟树突标记物微管相关蛋白（MAP2）的上调以及GnRH的表达和释放。提取物的氯仿部分被证明具有所有生物活性特性，因为它诱导了GnV - 3细胞的分化并上调了GnRH和MAP2的表达，类似于南非醉茄提取物。在南非醉茄水提取物（ASH - WEX）和氯仿部分中发现了睡茄酮和沃替西汀A，而睡茄酮是氯仿部分的主要成分。对成年雄性动物的初步体内研究表明，ASH - WEX能够上调GnRH水平，尽管不显著。综上所述，这些数据表明用南非醉茄提取物处理后GnV - 3细胞发生了显著的形态和生理变化，并可能暗示南非醉茄对体内生殖功能的潜在有益作用。
