Testicular Dysfunction Ameliorative Effect of the Methanolic Roots Extracts of Maytenus Procumbens and Ozoroa Paniculosa

Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2017, Article ID 8204816, 7 pages https://doi.org/10.1155/2017/8204816 Research Article Testicular Dysfunction Ameliorative Effect of the Methanolic Roots Extracts of Maytenus procumbens and Ozoroa paniculosa Nkosinathi David Cele,1 Nonhlakanipho Felicia Sangweni,1 Rebamang Anthony Mosa,1 Dambudzo Penduka,1 Geraldine Genevive Lazarus,2 Moganavelli Singh,2 Godfrey Elijah Zharare,3 and Andy Rowland Opoku1 1 Department of Biochemistry and Microbiology, University of Zululand, Private Bag X1001, KwaDlangezwa 3886, South Africa 2School of Life Sciences, Biochemistry Department, University of KwaZulu-Natal, Durban 4000, South Africa 3Department of Agriculture, University of Zululand, Private Bag X1001, KwaDlangezwa 3886, South Africa Correspondence should be addressed to Rebamang Anthony Mosa; [email protected] Received 12 July 2017; Revised 16 October 2017; Accepted 1 November 2017; Published 19 November 2017 Academic Editor: Carmen Mannucci Copyright © 2017 Nkosinathi David Cele et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The traditional use of medicinal plants in the management of ualsex dysfunctions has a long history. This study investigated testicular dysfunction ameliorative effect of the methanolic roots extracts of Maytenus procumbens and Ozoroa paniculosa in a butanol-induced testicular dysfunction rat model. The rats in respective experimental groups were orally administered with the extract at 50 and 250 mg/kg bw, daily for 28 days. The cytotoxicity of the extracts was evaluated against HEK293, MCF-7, and HT29 cell lines. The extracts exhibited moderate (LC50 30.3–330.2 g/mL) to weak (LC50 200.8–438.4 g/mL) cytotoxicity level on the cancer and normal cells, respectively. While relatively lower serum testosterone levels and total sperm count along with decreased numbers of spermatogonia were noted in the untreated group, all these parameters were improved in the groups treated with the extracts at 250 mg/kg. Improved histomorphological changes of the testes were also observed when compared to the untreated group. While the extracts (at 250 mg/kg) increased serum reduced glutathione content and decreased malondialdehyde content, a relatively higher serum creatinine level was also observed in the treated animals group. The results indicate that the two plant extracts have potential to ameliorate testicular dysfunction. 1. Introduction Despite an increasing number of men’s health clinics worldwide and the use of pharmaceutical agents to manage The maintenance of normal testicular function is crucial to complications of testicular dysfunction, substantial number male’s health. Testicular dysfunction is the main underlying of men, particularly living in rural areas, continue to consult cause of male developmental and sexual dysfunctions. It may traditional healers to improve their sexual functioning [7]. lead to a decline in fertility and androgen (testosterone) The use of medicinal plants, by traditional healers against deficiency in males. Testosterone deficiency has debilitating sexual dysfunctions, has a long and extensive history [8–10]. effects (hypogonadism, infertility, low libido, poor muscle Ethnopharmacological survey of traditional healers in North- strength, and erectile dysfunction) in the quality of life of men ern Kwa-Zulu Natal (South Africa) indicates that powdered [1, 2]. Various factors, either external or internal, are known to roots of Maytenus procumbens and Ozoroa paniculosa are compromise testicular functions. Such factors include expo- commonly used by traditional healers in the management of sure to toxic substances, diabetes mellitus, oxidative stress, testicular dysfunctions (Personal communication). and aging [3, 4]. The testicles are also vulnerable to oxidative Maytenus procumbens (L.F.) Loes (Celastraceae), com- stress due to their higher level of polyunsaturated fatty acids monly known as Dune Koko (English) and Umakhweni (PUFA) and presence of potential reactive oxygen species (Zulu), is a densely bush plant (about 6 m in height) native to (ROS) generating system [5, 6]. South Africa and predominant in the KwaZulu-Natal (KZN) 2 Evidence-Based Complementary and Alternative Medicine province.ItisalsofoundinSouthAmerica,NorthAfrica,and breast cancer (MCF-7) cell lines, using MTT assay [16]. The East Asia [11]. The leaf extract of M. procumbens has been cells were all obtained from the American Tissue Culture 4 reported to possess anticancer activity [11]. Ozoroa panicu- Collection (ATCC). The proliferating cells (1.8 × 10 cells/mL) ∘ losa (Sond.) R & A Femandes, (Anacardiaceae), commonly were exposed to the extract for 48 h at 37 C. The cells viability known as resin tree (English) and Umfico (Zulu), is an ever- was tested with the tetrazolium salt and after 4 h incubation ∘ green, semideciduous, small to medium sized single stemmed at 37 C, the formed formazan crystals were solubilised with tree (up to 9 m). The powdered stem bark of O. paniculosa is dimethyl sulfoxide (DMSO). The optical density of the used by Zulu traditional healers to treat acute inflammatory solutions was measured at 570 nm using a Mindray-96A condition of the chest [12]. This study is aimed at evaluating microplate reader. Percentage inhibition of cell viability was effect of methanolic roots extracts of M. procumbens and O. calculated using the formula paniculosa against testicular dysfunction in male rats. Effect (Ac − At) of the extracts on fertility is not covered in the current study. %celldeath=[ × 100] , (1) Ac 2. Materials and Methods whereAcistheabsorbanceofcontrolandAtistheabsorb- ance in the presence of extract. 2.1. Plant Material Preparation and Extraction. Fresh roots of M. procumbens and O. paniculosa were collected from 2.4. Effect of the Extract on Testicular Dysfunction in Rats. Ndumo and Mhlabayalingana, KZN, South Africa, in col- Approval for use of laboratory science animals and proce- laboration with a local traditional healer. The botanical dures was issued by the University of Zululand Research identification of the plants was confirmed by Dr. N.R Ntuli Ethics Committee (UZREC 171110-030 PGM 2015/198). and Dr. T.H.C Mostert at the Botany Department, Zululand Sprague-Dawley rats (150–200 g) of either sex were obtained University. The plants materials were air-dried and ground to from the animal unit of the Biochemistry Department, Uni- powder. Methanol was used to separately extract (1 : 5 w/v; versity of Zululand. Rats were kept under standard laboratory ∘ 48 h on a shaker at 150 rpm; room temperature) the powdered conditions (at 23 ± 2 C, 50 ± 5% humidity, and day-night plants materials. The filtrates were concentrated in vacuo ∘ cycle). The animals had access to fresh food and drinking (at 40 C). The obtained methanol-free crude extracts were ∘ water at leisure for the duration of the experiment. The rats safely stored at 4 Cuntiluse.Freshworkingsolutionswere were separated according to gender (maximum of four rats prepared by reconstituting the crude extract in distilled water per cage) and were allowed to acclimatize for at least five days just before use. before the experiment commenced. The effect of the extracts on testicular dysfunction was 2.2. Phytochemical Screening. Phytochemical analysis (Qual- evaluatedusingthebutanol-inducedtesticulardysfunctionin itative) of the plants material was performed following the rat model [17], with some modifications. The sexually active standard procedures described by Odebiyi and Sofowora [13]. male Sprague-Dawley rats received an intraperitoneal injec- The plants’ materials were screened for basic phytochemicals tion of n-butanolat25mg/kgbodyweightforfourdaysat such as tannins, alkaloids, saponins, steroids, flavonoids, and two-day intervals to induce testicular dysfunction. Four days terpernoids. after the induction of testicular dysfunction, the animals were mated with estrous female rats to establish their baseline sex- 2.2.1. Total Phenolic Content (TPC). The phenolic content of ual performance. The animals were then randomly divided the crude extracts was colourimetrically quantified follow- into seven groups (A–G) of five rats per group and put on ing the Folin-Ciocalteu reagent method [14]. Crude extract their respective treatments as shown as follows: (0.5 mg/mL) was well mixed with 1.5 mL of Folin-Ciocalteu reagent and 7.5% v/w sodium carbonate (1.2 mL). Following Group A: normal control group, received water (a 30 mins incubation, absorbance was read at 765 nm. The carrier solvent). total phenolic content of the plant extract was determined as Group B: testicular dysfunction induced (untreated gallic acid equivalent from a gallic acid calibration curve and group), received water only. expressed as mg/g dry plant material. Group C: testicular dysfunction induced, received sildenafil at 20 mg/kg. 2.2.2. Flavonoid Content (FC). The method described by Ordonez´ et al. [15] was followed to colourimetrically deter- Group D: testicular dysfunction induced, received mine the FC of the extracts. Crude extract (0.5 mg/mL) and 50 mg/kg M. procumbens. 0.5 ml of alcoholic aluminium chloride (2%) were well mixed Group E: testicular dysfunction induced, received ∘ together. After incubation (1 h at 25 C), absorbance was read 250 mg/kg M. procumbens. at 420 nm. The total flavonoid content of the extract was

Load more