تقييم فعالية المستخلص الكحولي والنانوي لنبات الهليون Asparagus officinalis في بعض المعايير الفسلجية والنسجية للجهاز التكاثري في ذكور الجرذان البيض المعاملة بـ Doxorubicin

Evaluation of the Efficacy of the Alcoholic and Nano Extract of Asparagus officinalis Plant on Some Physiological and Histological Parameters of the Reproductive System in Male Albino Rats Treated with Doxorubicin

The current study aimed to determine the protective efficacy of Asparagus officinalis plant, which was prepared as both an alcoholic and a nano extract synthesized zinc oxide against doxorubicin-induced reproductive toxicity in male albino rats by studying some physiological parameters and histological changes in the testes and epididymis.
The study was conducted the laboratories of the College of Education for Pure Sciences – Department of biology / University of Karbala and the animal house of the College of Pharmacy / University of Karbala and the laboratories of Al-Fadhil Foundation / Babylon and Al-Amin Center/ Al Najaf Al-Ashraf for the period from 25/3/2024 to 25/11/2024.
The study included (84) adult rats with average weights ranging between (225-250) grams and ages of (12-14) weeks, which were used in three experiments: two preliminary experiments and one main experiment. The first experiment aimed to determine the most effective concentration among three safe concentrations of the alcoholic extract of the Asparagus plant. It lasted 30 days, and 24 adult male rats were used,. They were randomly distributed into four groups of six rats each. The first group was considered the control group and was given ordinary water. As for the other three groups, each group was orally dosed with one of the three concentrations, respectively, of the alcoholic extract of the Asparagus plant (400, 500, 600 ) mg/kg of body weight After the experiment was completed, blood samples were drawn to estimate the activity of the enzyme superoxide dismutase (SOD), which was adopted as a standard to determine the most effective concentration of the alcoholic extract of the Asparagus plant ,the concentration of (500 ) mg/kg of body weight proved to be the most effective.
The second experiment was designed after the most effective concentration of the alcoholic extract of the Asparagus plant was determined according to the results of the first experiment. Then it was synthesized using zinc oxide, the nano extract was placed in three concentrations, respectively (125, 250, 375) mg/kg of body weight, using the same mechanism as the first experiment, and the activity of the enzyme (SOD) was measured, which was adopted as a standard to determine the most effective concentration of the nano extract of the Asparagus plant, the concentration of (250 ) mg/kg of body weight proved to be the most effective.
The main experiment was designed to study the effectiveness of the alcoholic and nano extract of a the Asparagus plant based on the results of the first and second experiments. It took (60) days, and (36) rats of the same ages and weights above were used. They were randomly divided into six groups, with six animals for each. The first group (G1) was given ordinary water and was considered the negative control group. The second group (G2) was injected intraperitoneally with doxorubicin at a dose of 2 mg/kg of body weight once every week. The third group (G3) was given alcoholic extract of Asparagus plant orally daily (500 mg/kg) of body weight, the fourth group (G4) was given alcoholic extract of Asparagus plant orally daily (500 mg/kg) and after 2-4 hours was injected with doxorubicin (2 mg/kg) of body weight once every week, the fifth group (G5) given nano extract of the Asparagus plant orally daily (250(mg/kg of body weight, the sixth group (G6) given nano extract of Asparagus plant orally daily (250 )mg/kg of body weight, and after (2-4) hours it was injected with doxorubicin (2) mg/kg of body weight once every week.
The weights of the animals were recorded at the beginning and end of the experiment for the purpose of calculating the rate of weight gain. The animals were sacrificed, and blood samples were collected to measure the following parameters: Malondialdehyde (MDA), Glutathione (GSH ) Catalase (CAT) , Testosterone hormone(T) , Follicle stimulating hormone (FSH) ,Interstitial cell stimulating hormone (ICSH), Total cholesterol (TC), Triglycerides (TG) , High- density lipoprotein( HDL) , and low – density lipoprotein ( LDL), The testes and epididymis were also removed for the purpose of calculating their average weights and were also used in the study of sperm parameters which included measuring sperm concentration and the percentages of sperm motility, vitality, and abnormal sperm, measuring the average diameters of the seminiferous tubules and their cavities, the average height of the germinal layer, the average diameters of the spermatogenic cells, which included : spermatogonia, primary spermatocytes, spermatids, and Sertoli cells , the average diameters of the epididymis and their cavities, and the average height of the epididymis epithelium.
The results of the first experiment indicated a significant increase (p≤0.05) in the level of SOD in G3 (500 ) mg/kg of body weight and G4(600 ) mg/kg of body weight Compared with G1, G3 was the best, While in the G2 (400 ) mg/kg of body weight there were no significant differences (p≥0.05) in the level of SOD compared with G1. The results of the second experiment showed a significant increase (p≤0.05) in the level of SOD in G2 (125 ) mg/kg of body weight and G3(250) mg/kg of body weight compared to G1, G3 had the best effect on the level of SOD, while in G4 (375) mg/kg of body weight there was a significant decrease (p≤0.05) in the level of SOD compared to G1.
The results of the main experiment showed a significant decrease (p≤0.05) in the weight gain rate, average weight of testes and epididymis, GSH level, CAT, reproductive hormone levels (T, LH, FSH), sperm concentration, sperm vitality, sperm motility, and HDL level , Average diameters of spermatocytes (spermatogonia, primary spermatocytes, spermatoblasts, Sertoli cells), average diameters of seminiferous tubules and epithelial cell layers and average diameters of epididymis and epididymis epithelium in the drug-treated group when compared to G1. As for the groups that were given the alcoholic and nano extract of the Asparagus plant , G3 and G5 and G4, G6, which were given the alcoholic and nano extract, respectively, with the drug, a significant increase (p≤0.05) was observed in the above parameters when compared with the positive control group G2. A significant increase (p≤0.05) was also observed in the levels of MDA, TG, TC, LDL, aneuploidy sperm, seminiferous tubule lumen diameter, and epididymis lumen diameter in the positive control group G2 compared to the negative control group G1. and a significant decrease (p≤0.05) in the same parameters in G3, G5, G4, and G6 compared to G2.
Histological examination results showed pathological changes in the testicular tissues of the G2 drug group, represented by the destruction of the basement membrane of the seminiferous tubule, atrophy of the seminiferous tubules, and the lining inside the seminal cavity being dilated and devoid of sperm. As for the epididymis tissues, the changes were represented by atrophy of the epididymis tubules and their absence or decrease in sperm, destruction of the cilia, and Presence of large interstitial spaces between the epididymis tubules, While groups G3 and G5 showed cohesion in the cells of the germinal layer and were more dense and differentiated from each other, the cavities of the seminiferous tubules appeared filled with spermatozoa. The nanometric extract showed the best positive effects on the testicular and epididymal tissues, while groups (G4, G6) showed an effective protective effect against the drug’s toxicity ,and the severe changes of the drug decreased, especially in G6.
We conclude from the above that the concentration (500 ( mg/kg of body weight of the alcoholic extract and the concentration (250) mg/kg of body weight of the nano-extract of the Asparagus plant were the best in their effectiveness against the toxic effects caused by the drug at a concentration of (2 ) mg/kg of body weight on the male reproductive system. Nanotechnology has also proven effective in improving the bioavailability of active compounds, thus increasing their antioxidant and anti-inflammatory effectiveness to perform the required functions and provide the necessary protection.