Effect of benzene extract of Ocimum sanctum leaves on cauda epididymal spermatozoa of rats.

BACKGROUND: Recent studies have shown that benzene extract of Ocimum sanctum (O. sanctum) leaves induces the ultrastructural changes in the epithelial cells of the cauda epididymis, its subsequent recovery in the seminiferous epithelium and fertility of male albino rats. OBJECTIVE: Our aim was to investigate the effect of benzene extract of O.sanctum leaves on the cauda epididymal sperm parameters, morphology and their organelles at the ultrastructural level in albino rats. MATERIALS AND METHODS: Wistar male rats (n=20) were allocated into two groups of control (n=10) and test group (n=10). The test group received benzene extract of O.sanctum leaves (250mg/kg/day) for 48 consequence days. Five animals from each group were used for fertility test. Twenty-four hours after the last dose, the rest of the control (n=5) and treated (n=5) animals were sacrificed by cervical dislocation and then the cauda epididymal plasma was used for sperm analysis, scanning electron microscopy (SEM) and transmission electron microscopic (TEM) studies. RESULTS: Sperm analysis of test group exhibited significant (p≤0.001) decrease in the sperm count, motility, speed and increase in sperm anomalies when compare to control group. SEM and TEM observation in treated animals indicated the morphological changes in plasma membrane as well as in the acrosomal membrane of spermatozoa, formation of a balloon-like cytoplasmic droplet in the mid-region of abnormal tail and disorganization or degeneration of mitochondria of sperm mitochondrial sheaths. CONCLUSION: The effects observed in this study may have resulted from a general alteration in the cauda epididymal milieu, probably due to androgen deficiency consequent to the anti-androgenic property of O.sanctum leaves.

Ultrastructural changes in Leydig cells and cauda epididymal spermatozoa induced by Azadirachta indica leaves in albino rats.

The effects of Azadirachta indica leaves (500 mg/kg body weight, orally/day), testosterone (0.25 mg/kg body weight/day, intramuscularly) and a combination of these two were investigated as to whether Azadirachta indica leaves affect the morphology of Leydig cells and epididymal sperm and their organelles at the ultrastructural level. Azadirachta indica treated rats showed atrophic seminiferous tubules with widening intercellular spaces. Leydig cells exhibited characteristics of degeneration such as indented nuclei; the commencement of degeneration was evident from less chromatin; the reduced amount of a marked decrease in organelle content and scarcity of other cell types of the interstitium in highly vacuolated cytoplasm, which were seem from both light and electron microscopic observations. The number of Leydig cells and their nuclear diameter were reduced significantly. Pathological changes in the spermatozoa of the cauda epididymis were observed and the spermatozoa retained cytoplasmic droplets. It is suggested that regression of Leydig cells and the absence of interstitium cell types indicates an androgen deficiency, which affects the spermatozoa in the epididymis by disturbing the internal epididymal milieu. The antiandrogenic and antispermatogenic properties of Azadirachta indica leaves appear to lead to a reduced fertilizing ability of the sperm.

Effect of graded doses of Momordica charantia seed extract on rat sperm: scanning electron microscope study.

Morphological changes in sperm of albino rats observed under scanning electron microscopy illustrate the disturbance in the plasma membrane as well as in the acrosomal membrane on treatment with effect of graded doses of alcohol seed extract from Momordica charantia. Considerable changes in the shape and size of the sperm head were observed, with the middle region of the sperm head being slightly constricted dorsoventrally. Most sperm appeared morphologically abnormal in the mid-region of the tail, with formation of a balloon-like cytoplasmic droplet. The results of this study suggest that such effects may have resulted from a general disturbance in proteins and an alteration in the cauda epididymal milieu, probably due to androgen deficiency consequent to the anti-androgenic property of Momordica charantia seeds.

Changes in Sertoli cells of albino rats induced by Azadirachta indica A. Juss leaves.

Azadirachta indica leaf powder, 0.05 mg testosterone, or combined leaf powder + testosterone was administered for 48 days to different groups of 3-month-old albino rats. Twenty-four hours after the final dose, the treated animals received mild ether anesthesia and the testes were dissected out and processed for light and electron microscope studies. Animals treated with A. indica leaf powder showed damaged tubules and exhibited an abundance of vacuoles, including inter-cellular spaces and intra-epithelial vacuoles of varying size in the cytoplasm of Sertoli cells. Upon treatment with A. indica leaf powder, bridges between Sertoli cells-Sertoli cells and Sertoli cell-germ cells were disturbed, coupled with changes in the Sertoli cells and cytoplasm along with its organelles. We suggest that the anti-androgenic property of A. indica leaves probably affects Sertoli cells, followed by the degeneration of germ cells, resulting in exfoliation.

Ultrastructural changes in cauda epididymidal epithelial cell types of Azadirachta indica leaf treated rats.

To assess if cauda epididymis is a target for the effect of A. indica leaves, Wistar strain male albino rats were administered (po) A. indica leaves (100 mg/rat/day for 24 days). Transmission electron microscopic analysis revealed that in the cauda epididymal epithelium the nuclei of principal cells were enlarged and the number of coated micropinocytotic vesicles of the apical cytoplasm decreased. Microvilli were missing and mitochondrial cristae and Golgi complex were highly disrupted. The cytoplasm was abounding with lysosomal bodies. The clear cells increased in perimeter and their nuclei increased in size and contained lesser chromatin. The nuclear membrane bulged out. The cytoplasm was vacuolized. Further, there was decrease in size of the lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and there was accumulation of lysosomal bodies. The changes in the principal and clear cells appear to be due to the effect of the hypoandrogen status caused by treatment with A. indica leaves and a direct action on the epididymal epithelium.

Azadirachta indica adversely affects sperm parameters and fructose levels in vas deferens fluid of albino rats.

Azadirachta indica treatment for 24 days in albino rats resulted in a decrease in the total sperm count, sperm motility, and forward velocity in vas deferens fluid. The percentage of abnormal sperm increased and the fructose content decreased. As diminished levels of fructose parallel androgen deficiency, we conclude that reduced androgen levels resulting from the anti-androgenic property of A. indica leaves probably influences the physiological maturation of sperm.

Reversible anti-fertility effect of benzene extract of Ocimum sanctum leaves on sperm parameters and fructose content in rats.

Treatment of albino rats with a benzene extract of Ocimum sanctum leaves (250 mg/kg body weight) for 48 d decreased total sperm count, sperm motility, and forward velocity. The percentage of abnormal sperm increased in caudal epididymal fluid, and the fructose content decreased in the caudal plasma of the epididymis and the seminal vesicles. The results suggest that such effects are due to androgen deprivation, caused by the anti-androgenic property of O. sanctum leaves. The effect was reversible because all parameters returned to normal 2 wk after the withdrawal of treatment.

Ultrastructural changes induced by leaves of Azadirachta indica (Neem) in the testis of albino rats.

The present work was designed to study the effect of Azadirachta indica (Neem) powder on rat testis using the electron microscope. Male albino rats received 100 mg each A. indica leaf powder orally (by gavage). On alternate days, a second group of rats received 0.125 mg testosterone dipropionate intramuscularly. A third group received both A. indica leaf powder by gavage and testosterone dipropionate intramuscularly. Suitable controls were maintained. After autopsy, ultrastructural analysis of the testis revealed that animals treated with testosterone dipropionate showed well-developed Sertoli cells and germ cells with well-developed cytoplasmic organelles. By contrast, in A. indica-treated rats, intracellular spaces and vacuolization were observed in Sertoli cells; whereas in Leydig cells, cytoplasmic inclusions appeared diminished, and the configuration of granular endoplasmic reticulum appeared as a single unbranched tubule. In late spermatids, defects were observed in the mitochondrial sheath. The ultrastructural changes seen in the A. indica-treated group provide a clue that A. indica leaves might affect spermatogenesis through antispermatogenic and antiandrogenic properties.