تقييم الدور الوقائي للمستخلص الكحولي لنبات Alpinia officinarum ضد السمية التكاثرية المستحثة بـ Bisphenol في ذكور الجرذان البيض

Evaluation of the protective role of alcoholic extract of Alpinia officinarum plant against Bisphenol-induced reproductive toxicity in male

This study was conducted at the Department of Biology/College of Education for Pure Sciences/University of Karbala for the period from February 2022 to January 2023. It included the phytochemical analysis of the alcoholic extract of the rhizomes of Lesser Galangal plant (Alpinia officinarum) and the evaluation of its protective role against reproductive toxicity induced by BPA in male albino rats.
The study included two experiments, the first aimed at determining the most effective concentration out of three safe concentrations, in which 24 adult male rats of the Ratus ratus strain, aged 14 weeks, weighed between 200-220 g , were used. They were randomly distributed into four groups (6 rats per group): the first was left without treatment and counted as a control group, while the other three groups were adminstrated orally with one of the safe concentrations of alcoholic extract (100, 200, 400 mg/kg/day), then after one hour they were given a single dose of BPA (50 mg/kg) for a period of 30 days, After that, blood samples were taken to estimate SOD activity, which was adopted as a criterion for selecting the most effective concentration.
The second experiment was designed to evaluate the protective role of the extract at the concentration that was determined according to the results of the first experiment, as it used 30 adult male rats (of the same ages and weights above) distributed randomly into five groups (6 rats per group): the first was left without treatment and was counted as a control group, while The second, third and fourth groups were dosed orally for 60 days with olive oil, galangal extract (400 mg/kg), bisphenol A (50 mg/kg) respectively, while the fifth group was given galangal extract and then adminstrated after one hour with bisphenol A and was counted as a protection group.
The weights of the animals were recorded at the beginning and end of the experiment, and the animals were sacrificed 24 hours after the last dose. Blood samples were collected for the purpose of conducting biochemical tests (Serum levels of testosterone (T), interstitial cell stimulating hormone (ICSH), follicle-stimulating hormone (FSH), malondialdehyde (MDA), glutathione (GSH), and catalase (CAT) activity) and the testes and epididymis were removed for use in the study of other criteria (sperm parameters, histological changes, assessment of deoxyribonucleic acid (DNA) damage) after they were cleaned and weighed.
The inferential results showed that the alcoholic extract of of lesser galangal rhizomes contains alkaloids, steroids, terpenes, phenols, flavonoids, saponins and tannins, while the results of quantitative assessment showed that it has high concentrations of phenols and flavonoids. The results of the identification and separation of the bioactive compounds using GC-MS revealed the presence of a number of bioactive compounds that differed in their surface area and retention time.
The results of the first experiment revealed that there was a significant decrease (p<0.05) in the activity of the SOD enzyme in the animals of the two groups treated with the two doses of 100 and 200 mg/kg of the extract compared to the control group, which did not show a significant difference between them and the treatment group with the concentration of 400 mg/kg of the extract. These results led to adopting a concentration of 400 mg/kg in the second experiment to evaluate its protective role against reproductive toxicity induced by BPA.
The results of the second experiment showed a significant decrease (p<0.05) in the average absolute weight of testes and epididymis, sperm concentration in testis and epididymis cauda, percentage of sperm motility and viability, level of reproductive hormones (T, ICSH, FSH), level of GSH and CAT activity, numbers of germ cells and sertoli cells, diameters of seminiferous tubules and the thickness of the germinal layer, diameters of epididymis cauda tubules and the thickness of the epithelial tissue lining them, and a significant increase (p<0.05) in the percentage of abnormal sperm, level of MDA, number of sperms that showed the shape of comet and the percentage of caudate content of fragmented deoxyribonucleic acid in animals of the BPA-treated group compared to the control group, while no significant difference was observed (p<0.05) in the amount of body weight gain between the two groups.
On the other hand, the results showed a significant increase (p<0.05) in the average absolute weight of testes and epididymis, sperm concentration in testis and epididymis cauda, percentage of sperm motility and viability, T, ICSH and FSH levels, GSH level and CAT enzyme activity. Number of spermatogenic cells and Sertoli cells, diameters of seminiferous tubules and thickness of germinal layer, diameters of tubules of the caudal epididymis and thickness of epithelial tissue lining them, and a significant decrease (p<0.05) in the percentage of abnormal sperms, MDA level, numbers of sperms that showed cometary shape and the percentage of tail content of fragmented DNA in the animals of the prevention group (the fifth group) compared to the control group, and there was no significant difference (p>0.05) in the amount of body weight gain between the two groups.
The results also revealed a significant increase (p<0.05) in the amount of body weight gain, testes and epididymis weights, sperm concentration in the testis and epididymis cauda, the level of T and FSH, the number of spermatogenic cells and Sertoli cells in the animals of the third group treated with the extract alone compared to a control group, while there was no significant difference (p>0.05) in the other criteria studied between the two groups.
changes in the tissues of the testis and epididymis cauda in the animals of the BPA-treated group compared with the control group, including: widening of the spaces between the seminiferous tubules, separation or sloughing of the germinal layer from the basement membrane with its disintegration significantly, the state of association between spermogenic cell and sertoli cells was decreased , reduction of the amount of sperm in the lumens of most of the seminiferous tubules and some of them being completely empty, it also included irregular distribution of epididymal tubules with disintegration and widening of the connective tissue between them, separation of the epithelial tissue lining the epididymal tubules from the basement membrane in some areas, reduction of sperm numbers in most epididymal tubules and some of them are empty. On the other hand, the histological sections of the animals of the prevention group showed a significant regression in the histological changes observed in the BPA-treated group.
It was concluded from the current study that the alcoholic extract of the rhizomes of Lesser Galangal plant at a concentration of 400 mg/kg has the ability to reduce the harmful effects on the male reproductive system resulting from exposure to BPA at a concentration of 50 mg/kg and provide the necessary protection to carry out the required functions through its direct effect and antioxidant activity.