BioFood Centre, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Nitra, Slovak Republic.
Physiol Res. 2021 Aug 31;70(4):615-625. doi: 10.33549/physiolres.934675. Epub 2021 Jun 1.
Several plants have the potential to protect essential reproductive processes such as spermatogenesis or steroidogenesis, however, effective concentrations and main mechanisms of action are still unknown. This in vitro study was aimed to assess the effects of Apium graveolens L., Levisticum officinale, and Calendula officinalis L. extracts on the structural integrity, functional activity and gap junctional intercellular communication (GJIC) in mice Leydig cells. TM3 cells were grown in the presence of experimental extracts (37.5; 75; 150 and 300 µg/ml) for 24 h. For the present study, high-performance liquid chromatography analysis was used to quantify flavonoids or phenolic acids. Subsequently, Leydig cell viability was assessed by alamarBlue assay, while the cell membrane integrity was detected by 5-carboxyfluorescein diacetate-acetoxymethyl ester. The level of steroid hormones production was determined by enzyme-linked immunosorbent assay. Additionally, GJIC was assessed by scalpel loading/dye transfer assay. According to our results, Apium graveolens L. significantly increased the viability and cell membrane integrity at 75 µg/ml (109.0±4.3%) followed by a decline at 300 µg/ml (89.4±2.3%). In case of Levisticum officinale and Calendula officinalis L. was observed significant decrease at 150 µg/ml (88.8±11.66%; 87.4±6.0%) and 300 µg/ml (86.2±9.3%; 84.1±4.6%). Furthermore, Apium graveolens L. significantly increased the progesterone and testosterone production (75 and 150 µg/ml) however, Levisticum officinale and Calendula officinalis L. significantly reduced steroid hormones synthesis at 150 and 300 µg/ml. Finally, the disturbance of GJIC was significantly affected at 300 µg/ml of Levisticum officinale (82.5±7.7%) and Calendula officinalis L. (79.8±7.0%). The balanced concentration ratio may support the Leydig cell function, steroidogenesis as well as all essential parameters that may significantly improve reproductive functions.
几种植物具有保护精子发生或类固醇生成等重要生殖过程的潜力，但有效浓度和主要作用机制尚不清楚。本体外研究旨在评估葛缕子、独活和金盏花提取物对小鼠睾丸间质细胞结构完整性、功能活性和缝隙连接细胞间通讯（GJIC）的影响。TM3 细胞在实验提取物（37.5；75；150 和 300μg/ml）存在的情况下培养 24 小时。目前的研究使用高效液相色谱分析来定量黄酮类化合物或酚酸。随后，通过 alamarBlue 测定法评估睾丸间质细胞活力，而通过 5-羧基荧光素二乙酸酯-乙酰氧基甲酯检测细胞膜完整性。通过酶联免疫吸附试验测定类固醇激素的产生水平。此外，通过手术刀加载/染料转移试验评估 GJIC。根据我们的结果，葛缕子在 75μg/ml 时显著增加了活力和细胞膜完整性（109.0±4.3%），随后在 300μg/ml 时下降（89.4±2.3%）。独活和金盏花在 150μg/ml（88.8±11.66%；87.4±6.0%）和 300μg/ml（86.2±9.3%；84.1±4.6%）时观察到显著下降。此外，葛缕子显著增加了孕酮和睾酮的产生（75 和 150μg/ml），然而，独活和金盏花在 150 和 300μg/ml 时显著降低了类固醇激素的合成。最后，在 300μg/ml 时，独活（82.5±7.7%）和金盏花（79.8±7.0%）显著影响了 GJIC 的干扰。平衡的浓度比可能支持睾丸间质细胞功能、类固醇生成以及所有重要参数，这些参数可能显著改善生殖功能。
