Andrologia. 2020;00:e13825. wileyonlinelibrary.com/journal/and | 1 of 8 https://doi.org/10.1111/and.13825 © 2020 Wiley-VCH GmbH 1 | INTRODUCTION Moringa oleifera (MO) belonging to the genus Moringa in the fam- ily of moringaceae is commonly known as ben oil tree, benzoil tree, drumstick tree, horseradish tree or Moringa (Fahey et al., 2019). It is native to the sub-Himalayan region of North-West India, Pakistan, Bangladesh and Afghanistan and is cultivated in Africa, Arabia, South East Asia, South America, Pacific and Caribbean Islands (Iqbal & Bhanger, 2006; Melo, Vargas, Quirino, & Calvo, 2013; Sreelatha & Padma, 2009). The leaves, seeds and pods of MO are widely used by humans for medicinal purposes to cure a variety of diseases (Gopalakrishnan, Doriya, & Kumar, 2016). The leaves of MO have significant antioxidant activity (He et al., 2018; Moyo, Oyedemi, Masika, & Muchenje, 2012) as a result of its ascorbic acid (Vitamin C), β-carotene, quercetin, kaempferol and phenolic acids content (Anwar, Latif, Ashraf, & Gilani, 2007; Gupta et al., 2012; Kumar, Pandey, Mohan, & Singh, 2012; Mahajan, Mali, & Mehta, 2007; Sreelatha, Jeyachitra, & Padma, 2011). Traditionally, MO is used to enhance male sexual functions such as libido, improve sperm quality and as an aphrodisiac in the treat- ment of sexual dysfunction (Lalas & Tsaknis, 2002; Prabsattroo, Wattanathorn, Iamsaard, Muchimapura, & Thukhammee, 2012). For instance, studies have shown increased weight of male reproductive organs, sperm parameters, luteinising hormone (LH), follicle-stimu- lating hormone (FSH), testosterone levels, as well as enhanced sex- ual activities in rats (Dafaalla, Abass, Abdoun, Hassan, & Idris, 2017; Ogunsola, Joshua, Sunday, Nl, & As, 2017; Prabsattroo et al., 2012; Zade, Dabhadkar, Thakare, & Pare, 2013). In mice, however, an Received: 20 June 2020 | Revised: 29 July 2020 | Accepted: 9 August 2020 DOI: 10.1111/and.13825 ORIGINAL ARTICLE Androgenic effect of aqueous leaf extract of Moringa oleifera on Leydig TM3 cells in vitro Chinyerum S. Opuwari 1 | Matome N. Matshipi 2 | Mantaneng K. Phaahla 2 | Mmaphulane A. Setumo 2 | Rantobeng T. Moraswi 2 | Amukelani A. Zitha 2 | Ugochukwu Offor 1 | Solomon S. R. Choma 2 1 Department of Pre-Clinical Sciences, University of Limpopo, Polokwane, South Africa 2 Department of Pathology and Medical Sciences, University of Limpopo, Polokwane, South Africa Correspondence Chinyerum S. Opuwari, Department of Pre-Clinical Sciences, University of Limpopo, University road, Polokwane, 0727, South Africa. Email: chinyerum.opuwari@ul.ac.za Funding information The University of Limpopo supported this study. Abstract Moringa oleifera (MO) is an excellent source of dietary antioxidant. MO is used tra- ditionally to enhance libido and as an aphrodisiac in the treatment of sexual dys- function. This study aimed to investigate the direct effect of aqueous leaf extract of MO on Leydig cell in vitro. Specifically, the effect of MO on viability, testosterone production, antioxidant activity and lipid peroxidation on TM3 cells were evaluated. TM3 cells seeded for 24 hr were exposed to aqueous leaf extract of MO (0, 10, 50, 100, 250, 500 and 1,000 µg/ml) for 24 hr, in the absence or presence of human chori- onic gonadotropin (hCG; 6 mIU/200 µl). Cell viability remained unchanged while tes- tosterone production significantly increased at 500 and 1,000 µg/ml of the extract under stimulatory conditions by 34 and 45% respectively. Glutathione level sub- stantially increased at 250 µg/ml, while lipid peroxidation, catalase and superoxide dismutase activity, and total antioxidant capacity remained unchanged. Our results demonstrate the androgenic effect of MO especially at high concentrations in TM3 cells. The androgenic effect may be attributed to its antioxidant enzyme activities. KEYWORDS antioxidant, fertility, Moringa oleifera, oxidative stress, testosterone