دراسة نسجية وكيميائية مناعية وفسلجية لتقييم دور المستخلص المائي والنانوي لجذور نبات الماكا في تحسين كفاءة الجهاز التناسلي في ذكور الجرذان البيض المعاملة بعقار الأوكسي ميثلون

Histological, immunochemical and physiological study to evaluate the role of aqueous and nano extracts of Maca roots oxymethylone

The present study aimed to know the effect of aqueous and nano extract of the roots of Maca plant Lepidium meyenii on the histological changes in the testes, epididymis and prostate and some blood biochemical parameters in male white rats treated with Oxymetholone (OX). The present study showed the chemical content of the aqueous extract of the roots of Maca plant where the compounds were identified using Gas Chromatography Mass Spectrometry (GC-MS) technique and it was found that there were ten chemical compounds: Dodecane, cyclohexasiloxane -dodecamethyl, Diethyl phthalate, Cedrandiol, pathalic acid, butyl tetradecyl ester, Methyl palmitate, Methyl linoleate, Oleic acid, methyl ester, palmitic acid, methyl ester and these compounds have an important and effective role in medical treatments.
The current study was conducted in the animal house of the College of Pharmacy / University of Karbala for 55 days from December 2023 to the middle of February 2024. 42 adult male white rats were used with an average weight of (250-350) g and an age of approximately (12-14) weeks. The rats were randomly distributed into six groups with (seven animals per group). The first group (G1) was considered the negative control group, which was given water orally for 55 days. The second group (G2) was dosed with Oxymetholone at a concentration of 5 mg/kg of body weight throughout the experiment and was considered the positive control group. The third group (G3) was dosed orally with the aqueous extract of the roots of the Maca plant Lepidium meyenii at a concentration of (200) mg/kg of body weight, respectively, daily for 55 days). The fourth group (G4) was dosed orally with the extract Nano extract of maca roots at a concentration of (100 mg/kg) of body weight, the fifth group was orally dosed with the aqueous extract of maca roots at a concentration of (200 mg/kg) and treated with Oxymetholone at a concentration of (5 mg/kg), respectively, while the sixth group (G6) was orally dosed with the nano extract of maca roots at a concentration of (100 mg/kg) of body weight and treated with Oxymetholone at a concentration of (5 mg/kg) four hours after dosing with Oxymetholone.
Blood samples were collected for the above groups after 55 days of dosing. Histological sections of the testes, epididymis and prostate were taken for the purpose of studying the histological changes on them, which include: measuring the average diameters of each of the seminiferous tubules and their cavities, the average thickness of the germinal layer of the seminiferous tubules, measuring the average diameters of each of the epididymis and their cavities, and the average height of the epididymal epithelium. Blood serum was obtained for the purpose of measuring the following biochemical parameters: levels of antioxidants malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), measuring the level of testicular lipid hormone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH), as well as performing the technique Immunohistochemistry of both testicular and epididymal tissues and sperm concentration in the tail of the epididymis was measured.
Daily administration of Oxymetholone for 55 days led to histological changes in the testicles, represented by degeneration of the germinal layer and disintegration of its cells, with separation of the germinal layer from the basement membrane in most of the tubules, with interstitial spaces between the seminiferous tubules. Sperm were few or absent in the tubule cavities, with reduction and disintegration of the interstitial tissue, as well as a lack of sperm cells and a decrease in the size of the germinal layer with cell degeneration. A significant decrease (p≤0.05) was observed in the rate of elevation of the epididymal epithelium and the rate of diameters of each of the seminiferous tubules, as well as the diameter of the lumen of the seminiferous tubules, compared to the negative control group. As for the histological changes of the epididymis, there was a decrease or absence of sperm in the lumen of the tubules, as well as tissue damage due to the destruction of the epithelial cells lining the tubules and the reduced epithelium, as well as a decrease in the presence of smooth muscles surrounding the tubules, with a significant decrease (P≤0.05) in the average diameters of the epididymis and their cavities and the average height of the epididymal epithelium. As for the histological changes of the prostate, there was a disintegration of the glandular tissue of the alveoli and the disintegrated connective tissue and the disintegration of the glandular tissue cells and a decrease in the glandular secretions of the alveoli compared to the G1. A significant decrease (P<0.05) was also observed in the average levels of GSH, the level of SOD, and MDA in the average levels of the hormones (T, LH, FSH) as well as in the average sperm concentration, while a significant increase (P≤0.05) was observed in the average levels of GSH, the level of SOD, and MDA in the average levels of the hormones (T, LH, FSH). as well as the sperm concentration rate compared to G1.
Oral administration of the aqueous extract of the roots of the Maca plant Lepidium meyenii at a concentration of (200 mg/kg) of body weight in G3 did not show any histological changes in the tissues of the testicle, epididymis and prostate, and there were no significant differences(p≥0.05) in the average diameters of each of the testicle and epididymis in G3 compared with G1. It was also noted that there was a significant increase (p≤0.05) in the average diameters of the seminiferous tubules transporting sperm, the diameter of the cavity and the thickness of the germinal layer of the testicular tissue of the G3 compared with G2, and also a significant increase (p≤0.05) in the concentration of (GSH, SOD) and the level of hormones including (T, LH, FSH) hormones, as well as in the concentration of sperm, with a significant decrease (p≤0.05) in the concentration of MDA compared with the G2.
Oral administration of the nano extract of the roots of the Maca plant Lepidium meyenii at a concentration of (100 mg/kg) of body weight did not cause any histological changes in the tissues of the testicle, epididymis and prostate, and there were no significant differences in the average diameters of each testicle. It was also noted that there was a significant increase (P≤0.05) in the average diameter of the epididymis and no significant differences in the average diameter of the epididymis cavity and the average thickness of the germinal layer compared to the G1. It was also noted that there was a significant increase (P≤0.05) in the level of GSH, and no significant differences in the level of (MDA, SOD) as well as in the level of the hormone (T, LH, FSH) as well as in the concentration of sperm compared to the G1.
The results of histological examination of the testicular tissue in the preventive groups treated with the aqueous and nano extract of Maca roots at a concentration of (100-200 mg/kg) and treated with the drug Oxymetholone at a concentration of (5 mg/kg) showed that the layers of cells forming sperm were observed in addition to the regularity of the seminiferous tubules and the presence of sperm in some of them with the presence of Leydig cells in the interstitial tissue and an increase in the thickness of the germinal epithelial layer forming sperm as well as a decrease in the diameter of the middle cavity and the cells that generate sperm inside the seminiferous tubules and sperm progenitors were observed in it, as the tissue appeared closer to the G1. As for the average diameters of the testicular tissue, a significant increase (P≤0.05) was observed in the average diameters of each of the seminiferous tubule, the diameter of the cavity and the thickness of the germinal layer compared to the G2. As for the epididymis tissue, the histological changes in the preventive groups were observed in it, where the normal structure of the epididymis with the tubules was observed. Regular epididymis and also filling the cavities with mature sperm and lack of smooth muscle cells around the tubules and the presence of interstitial spaces between the tubules of the epididymis compared to the G2. As for the average diameters of the epididymal tissue in the preventive groups (G5, G6), a significant increase (P≤0.05) was observed in the average diameters of each of the epididymis, the diameter of the lumen and the thickness of the germinal layer compared to the G2 . As for the histological changes of the prostate tissue in the preventive groups, it is noted that the connective tissue is regular and cohesive and the secretions are normal between the alveoli and the alveoli and glandular tissue appear normal.
The results of the current study in the preventive groups treated with the aqueous and nano extract of the roots of the Maca plant Lepidium meyenii at a concentration of (100,200 mg/kg) with the drug Oxymetholone showed a significant increase (p≤0.05) in the level of GSH, the level of SOD, and the level of each of the hormones (T, FSH, LH) as well as the concentration of sperm and a significant decrease (p≤0.05) in the level of MDA compared to the G2. The immunohistochemical study reinforced the results of the study and showed the brown color of the tissues indicating the presence of immune cytokines represented by tumor necrosis factor alpha (TNF_ἁ Tumar necrosis factor). We conclude from the current study the effectiveness of the nano extract of Maca roots at a concentration of (100) mg/kg, which had the strongest role in inhibiting the activity of free radicals and reducing oxidative stress induced by the drug Oxymetholone in the tissue of the testicle, epididymis, prostate and some functional parameters in male white rats.