Histological changes of extract cumin cyminum on the leydig cells, seminiferous tubules and epididymis in adult rabbits (Oryctolagus cuniculus)

1. This study focuses on the most significant histological alterations connected to seminiferous tubules.
2. Frequent cumin use inhibits Leydig cell activity.
3. Insufficient spermatogenesis as a result of consuming too much cumin in the diet.

Introduction

Materials and methods

Experimental design

In the current study, ten adult male local rabbits weighing approximately 1150-1300 g have always been separated into two groups (5 group). The control group received a typicaliet and water, whereas the treatment group received cumin extract 250 mg/kg b.w. through stomach tube feeding (24). For all groups, the experiment lasted for six weeks. By administering intramuscular injections of xylazine 0.5 ml and ketamine, all study rabbits were killed 0.5 ml.

Cumin extraction

Cumin powders were purchased from the Kerbala city market. Each 25 g of dry cumin powder was used to prepare an extract, after which it was treated with 10 ml of boiling distilled water. The extraction process required thirty minutes. Each extract had been filtered, and the content was modified so that each ml of sample was equal to 250 mg/ml. Those solutions are subsequently stored in a tightly sealed container (25).

Histological preparations

The testis served as the source of the anatomical samples. The samples, which were about 0.5 cm in size, were then preserved in 10% Neutral buffered formalin for 48 hours. Hematoxylin and eosin satin were used with conventional histological methods to distinguish distinct tissue components (26). The germinal layer of the seminiferous tubules and the interstitial space occupied by Leydig cells and all tissue sections have been incorporated into the histological measurements. The histological characteristics were analyzed with the help of a digital USB microscopic camera (Canon 550D, 18 Megapixel, Japan), which was attached to a Novel microscope. A stage micrometer has been employed to correlate the objective lenses with the program (27).

Ethical approve

Under the reference number UOK.VET.AN.2022.052, this research was carried out in the anatomical laboratory of the College of Veterinary Medicine at the University of Kerbala.

Results

Histological features

The illustration shows the typical characteristics of the testes in the control group, including regular histological features with well-developed spermatids and a germ cell lining performing cell proliferation. Furthermore, we observed Sertoli cell activity and their presence in their usual location in the basement membrane of seminiferous tubules, which revealed the arrangement of spermatogonial stages. The mean diameters of seminiferous tubules were approximately 98.14 µm (Figure 1). In the control group, Leydig cells were observed to be active; these cells appeared as a massive regular, with non-granular and spherical cytoplasm and abundant nuclei with visible nucleoli. The majority of the nuclei are located in the cytoplasm's middle. The mean thickness of the Leydig area was 152.21 µm (Figure 2). Moreover, the histological features of the rabbit epididymis duct in the control group were lined by pseudostratified columnar epithelium and a large number of spermatozoa in the epididymis lumen (Figure 3).

When the histological testis characteristics of the extract-cumin group were examined under the microscope, numerous changes were observed, including the destruction of the seminiferous tubules, a significant number of damaged cells, the absence of spermatozoa in the tubule lumen output, spermatogonia detaching from the basal membrane in some areas, and an increase in the space between the seminiferous tubules, the mean diameters of seminiferous tubule about 50.22 µm (Figure 4). In this Group, we observed the decrease in the number of Leydig cells an appearance of inactivity, limited cytoplasm, and psychotic nucleus the mean thickness of Leydig area was 92.45 µm (Figure 5). The epididymis duct appears empty in this group due to spermatozoa in the epididymis lumen (Figure 6).

Figure 1: Photomicrograph of control testis in rabbits showing the normal histological features in semiferoius tubules (yellow arrows), and activity secretion in interstitial space (blue arrows). H&E stain 10x.

Figure 2: Photomicrograph of control testis in rabbits showing the crowded and activity in leydig cells (black arrow). H&E stain 40x.

Figure 3: Photomicrograph of control epididymis in rabbits showing a lot of sperms in ducts of epididymis (black arrow), and activity lining epithelia (blue arrow). H&E stain 10x.

Figure 4: Photomicrograph of extract cumin testis in rabbits showing the damage in semiferoius tubules (black arrows). H&E stain 10x.

Figure 5: Photomicrograph of extract cumin testis in rabbits showing the leydig cells in active and smaller area size (black arrow). H&E stain 40x.

Figure 6: Photomicrograph of extract cumin epididymis in rabbits showing of ducts of epididymis from sperms (black arrow), and inactivity lining epithelia (blue arrow). H&E stain 10x.

Discussion

The current study shows the typical appearances of the testis in the control group, including regular histological features with very well-developed germ cell lining performing cell proliferation. In contrast, the current study found numerous modifications in the extract-cumin group, including destruction to the seminiferous tubules, a significant number of damaged cells, the absence of spermatozoa in the tubule lumen output, spermatogonia detaching from the basal membrane in some areas, and an increase in the space between the seminiferous tubules. This outcome is similar to that of Abdi et al. (28) and Taha (29), which mentions that in a healthy condition, spermatogonia and the basement membrane are nearby in the seminiferous tubules, where germ cells are alive and appear active. Unnoticed changes also included the degradation of specific spermatogenic cells and an accumulation in the tubular hollow. These significances might have resulted from excessive consumption of cumin extract, which is widely used as a body slimming agent and may have caused weight loss.

The control group observed regular histological features with the activity of Sertoli cells in the control group; additionally, the Leydig cells had nuclei located in the cytoplasm's middle, a wide interstitial area that was occupied by Leydig cells, whereas cumin describes a decrease in the number of Leydig cells, inactivity appearance, limited cytoplasm, and psychotic nuclei in the extract group. Findings are akin to Tripepi et al. (30) and Malhi et al. (31) those that describe the Leydig cells. They were found in groups or cords of various shapes and sizes with round nuclei and granular cytoplasm. These cells seem to be distributed randomly, some being perivascular, others related to vessels. The testicular hormone was markedly lower in these results compared to the control group, comparable to those of Willatgamuwa et al. (32), which evaluated the therapy with a water extract of the cumin plant. These studies, however, contradict Smith et al. (33), Hipler et al. (34) and Fonseca et al. (35) thatclaimed that the Cumin therapy was ineffective when administered alone in sperm characteristics and had a detrimental impact. Gupta et al. (36), Sharifi et al. (37) and Al-Hafedh et al. (38). The testicular tissue may have acquired changes due to all these histological changes caused by hormone changes. Consequently, these variations need the cells responsible for adapting these mechanisms to maintain the new metabolic balance.

The present study explains why the epididymis duct in the extract cumin team appeared to be vacant of spermatozoa, in contrast to the morphological features of the rabbit epididymis duct in the comparison group, which were lined by a significant number of spermatozoa in the epididymis lumen and had a regular appearance of pseudostratified columnar epithelium. Our findings are consistent with those of others who believed that some variables determined by sperm concentration are elements of sperm parameters that significantly affect male fertility Roshankhah et al. (39), Al-Allaf et al. (40) and Amin et al. (41). Herbal medicine may be widely used in many cultures to treat conditions like obesity and to enhance sperm quality. Hence, excessive use always has a harmful impact on the amount and quality of sperm production.

Conclusion

The College of Veterinary Medicine at the University of Kerbala is acknowledged for facilitating this investigation.

Conflict of interest

There is absolutely no apparent conflict, according to the author.