Ameliorations of Nigella sativa seed oil against endocrine glands histopathologies of Bifenthrin exposure in mice

Abstract

Histopathological and micrometric alterations of mice endocrine glands (hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas) against bifenthrin (Bf2.5 and Bf5) exposure were investigated. Additionally, the rescuing potential of Nigella sativa seed oil (Ns) were also investigated. The study comprise on 60 male albino mice (27-30g) - randomly distributed in 6 groups (n=10). The vehicle control group was maintained and treated with 0.1ml of corn oil only for 1 – 14 days. The positive control group was treated with 0.1ml of corn oil for 1- 7 days and 0.1ml of 10% Nigella sativa seed oil mixed in corn oil for next 8-14 days, whereas the (Bf2.5) and (Bf5) groups were treated with 2.5mg/kg and 5mg/kg Bf solutions in corn oil of the respective doses of insecticide for 1- 7 days followed by 0.1ml of corn oil on 8-14 days. The BfNs2.5 and BfNs5 group animals were treated with 0.1 ml of respective doses of Bf on 1-7 days followed by 10% Nigella sativa seed oil solution as in the positive control group. All animals were sacrificed on day 15 to recover the major endocrine glands (hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas). Results show various histopathological alterations in endocrine glands (hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas) related to Bf exposure. The characteristic pathologies of hypothalamus discovered in this study include focal degenerations in neuro-secretary tracts of neuroglial cells, enlargement of the neuro-secretary neuronal cell bodies primarily with the appearance of large vacuolations in the neuronal cytoplasm in Bf2.5 and Bf5 groups.The micrometric results support the histopathological findings and show significant variations in various micrometric parameters among the six groups. For example, Mean number of neurons per (2500 µm2 ) in hypothalamus remained significantly (p≤0.05) lower in Bf2.5 (4.14± 0.22), Bf5 (3.30±0.18),BfNs2.5 (5.04±0.20)and BfNs (54.22±0.23) as compared to control (6.56±0.34) and Ns (6.93±0.39) groups.Contrarily mean cross sectional area of neuroglial cells in hypothalamus was significantly (2) (p≥0.05)higher in Bf5 (86.43±3.53) as compared toNs (67.75±2.22) and control (68.89±2.07) group. In pituitary gland Bf exposure was found to induce histopathological changes i.e. cellular vacuolations causing hypertrophy finally leading to secondary atrophy and adenohypophyseal tissue damage in terms of focal degenerations in the typically arranged nest and cords of the endocrine cells. The cell nuclei were slightly elliptical instead of typical rounded appearance having less densely stained sinusoid.The supportive micrometric results also indicate significant variations in various parameters studied. Like, mean number of acidophils per unit area (2500µm2 ) of adenohypophysis were significantly lower(p≤0.05) in Bf2.5 (5.33±0.47), Bf5 (4.6±0.36) and BfNs5 (7.33±0.88) as compared to control (12.73±0.51) group.Similarly, number of chromophobes have shown significant decline (p≤0.05) inBf5 (0.00±0.00), and BfNs5 (0.00±0.00) as compared to control (1.46±0.13). Contrarily, the numbers of basophilic cells were significantly higher (p≥0.05) in Bf2.5(8.87±0.60), Bf5 (9.8±0.37), and BfNs5 (8.86±0.68) as compared to control (20.6±0.73). Likewise total no. of pituicytes/unit area were significantly lower(p≤0.05) in Bf2.5(15.26± 0.63), Bf5 (14.4±0.51) and BfNs5 (14.4±0.51) as compared to the control (20.6±0.73) group. In Bf treated groups dose dependent effect of thyroparathyroid histotoxicosis were observed that include depletion of thyroglobulin, fluid filled thyroid follicles, cellular necrosis, follicular atresia and fibrosis, degeneration of chief cells of parathyroid and parathyroid vacuolations.The micrometric data of thyroparathyroid for mean CSA of medial follicles indicated significant decline (p≤0.05)in Bf2.5(411.33±48.50), Bf5 (261.33±24.30), and BfNs5 (364±27.78) groups as compared to control (770.94±93.17). Similarly mean CSA of marginal follicles of thyroid was significantly lower(p≤0.05) in Bf5 (3850.2±171.4) group as compared to control (3803.74±179.6) group. Likewise, mean CSA of follicular cells has shown significant decline (p≤0.05) in Bf2.5(20.04±0.89), Bf5 (13.57±0.71) and BfNs5 (18.63±0.79) as compared to the control group.The micrometric data of parathyroid sections (3) also indicate significant (p<0.05) decline of CSA of chief cells inBf2.5(16.64±1.23), Bf5 (13.34±0.74), BfNs2.5 (23.70±0.69)and BfNs5 (19.42±0.67) as compared to control (29.59±0.67) and Ns (31.68±1.1) groups. The histopathological land marks of adrenal gland observed in the Bf treated groups include randomly located enlarged follicles with hypertrophy of the chromaffin cells; marks of discrete medullary endocrine cells apoptosis were also distinguishable. The zona glomerulosa remained more or less entirely unchanged yet the zona fasciculata exhibited cellular hypertrophy, moreover the breached ×-zone allowing profusion of blood in zona fasciculata caused interruption of the cortical fascicles at numerous places.The micrometric results support the histopathological findings and show significant enlargement (p<0.05) in the meancross-sectional area (CSA) of the chromaffin cells in the Bf2.5 (185±2.6) and Bf5 (192±2.6) groups compared to the BfNs2.5 (167±1.9) and BfNs5 (158±2.6 μm2) groups. Similarly, thenumber of cells per fascicle in the Bf2.5, Bf5, BfNs2.5, and BfNs5 groups (18.6±0.4,19.4±0.3, 16.3±0.5, and 16.6±0.4 respectively) were significantly higher (p<0.05) than inthe control and Ns groups (14.9±0.3 and 15.5±0.4, respectively). In contrast, the mean CSAof the fascicular cells was significantly higher (p<0.05) in the Ns (101±2) and control (103±2.8) groups than in the Bf treated groups (75±1.9, 66±2.5, 95±1.8, and91±2, respectively). In Bftreated groups, periphery of pancreatic islets give an impression of disassociation of the adjacent cells with focal endocrine cells degeneration. The islets seems to show enlargement of individual endocrine cells as their nuclei were seem more distantly placed as compared to the control group. Some apoptotic signs in the form of nuclear distortions and dissolutions were also seen.The micrometry estimations have likewise showed a significantly (p<0.05) higher mean Cross-Sectional Area (CSA) of the pancreatic islets in Ns (16521±1744), BfNs2.5 (16930±1700) and Bf2.5 (13703±1572) than the Bf5 (6382±1405), Vc (9464.75±955) and Bf Ns5 (11451±1099). The mean CSA of endocrine cells of islets in Bf2.5 (30.1±1.39µ2 ) remained significantly higher than rest of all the groups (4) (Vc: 23.1±0.8; Ns: 21.6±0.85; Bf5: 24.4±1.03; BfNs2.5: 25.2±0.95 and BfNs5: 22.1±0.71) and the mean number of cells in the islets/unit area (289µ2 ) in Bf2.5 and Bf5 (141.37±6.3 and 132.03±6.5 respectively) remained significantly lower than Ns (150.89±6.7), Vc (174.43±6.6), BfNs2.5 (172.97±6.4) and BfNs5 (150.61±6.5) groups. The architectural distribution and histometery of Ns group did not differ much with that of Vc group. However in BfNs2.5 and BfNs5 the pace of rehabilitation of histoarchitecture of endocrine glands were much higher than that of Bf2.5 and Bf5 groups, clearly indicating the significance of Ns seed oil in rehabilitation of histological land marks of intoxicated mice endocrine glands. The Nigella sativa seed oil post treated group has shown rehabilitative effects against histopathologies. On the basis of these findings, it is concluded that insecticide harbor endocrine gland histopathological potentials that are at least partially rescue-able by Nigella sativa oil treatment. These findings thus indicate effective neutraceutical potential of Ns oil for possible human consumption in similar accidental exposure to the insecticide. Histopathological and micrometric alterations of mice endocrine glands( hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas) against bifenthrin (Bf2.5 and Bf5) exposure were investigated, additionally, the rescuing potential of Nigella sativa seed oil (Ns) were also investigated. The study comprise on 60 male albino mice (28-30g) - randomly distributed in 6 groups (n=10). The vehicle control group was maintained and treated with 0.1ml of corn oil only for 1 – 14 days. The positive control group was treated with 0.1ml of corn oil for 1- 7 days and 0.1ml of 10% Nigella sativa seed oil mixed in corn oil for next 8-14 days, whereas the (Bf2.5) and (Bf5) groups were treated with 2.5mg/kg and 5mg/kg Bf solutions in corn oil of the respective doses of insecticide for 1- 7 days followed by 0.1ml of corn oil on 8-14 days. The BfNs2.5 and BfNs5 group animals were treated with 0.1 ml of respective doses of Bf on 1-7 days followed by 10% Nigella sativa seed oil solution as in the positive control group. All animals were sacrificed on day 15 to recover the major (5) endocrine glands (hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas). Results show various histopathological alterations in endocrine glands (hypothalamus, pituitary, thyroid, parathyroid, adrenal and pancreas) related to Bf exposure. The characteristic pathologies of hypothalamus discovered in this study include focal degenerations in neuro-secretary tracts of neuroglial cells, enlargement of the neuro-secretary neuronal cell bodies primarily with the appearance of large vacuolations in the neuronal cytoplasm in Bf2.5 and Bf5 groups. In pituitary gland Bf exposure was found to induce histopathological changes i.e. cellular vacuolations causing hypertrophy finally leading to secondary atrophy and adenohypophyseal tissue damage in terms of focal degenerations in the typically arranged nest and cords of the endocrine cells. The cell nuclei were slightly elliptical instead of typical rounded appearance having less densely stained sinusoid. In Bf treated groups dose dependent effect of thyroparathyroid histotoxicosis were observed that include depletion of thyroglobulin, fluid filled thyroid follicles, cellular necrosis, follicular atresia and fibrosis, degeneration of chief cells of parathyroid and parathyroid vacuolations.The histopathological land marks of adrenal gland observed in the Bf treated groups include randomly located enlarged follicles with hypertrophy of the chromaffin cells; marks of discrete medullary endocrine cells apoptosis were also distinguishable. The zona glomerulosa remained more or less entirely unchanged yet the zona fasciculata exhibited cellular hypertrophy, moreover the breached ×-zone allowing profusion of blood in zona fasciculata caused interruption of the cortical fascicles at numerous places.In Bftreated groups, periphery of pancreatic islets give an impression of disassociation of the adjacent cells with focal endocrine cells degeneration. The islets seems to show enlargement of individual endocrine cells as their nuclei were seem more distantly placed as compared to the control group. Some apoptotic signs in the form of nuclear distortions and dissolutions were also seen. Additionally significantalterations (p≤0.05) were observed in micrometric parameters of hypothalamus and pituitary include theCSA and number of neuroglial cells per (6) unit area of hypothalamus, number of acidophils, basophils, chromophobes, total number of pituicytes per unit area, CSA of pituicytes and relative area occupied by pituicytes. Similar alterations were observed in micrometric parameters of thyroparathyroid, adrenal and pancreas including the CSA of the medial and marginal follicles, follicular cells nuclei of thyroid and chief cells in parathyroid; mean CSA of cells in zona glomerulosa, zona fasciculate, chromaffin cells, chromaffin cell’s nuclei, cytoplasmic granules per chromaffin cells and non-granular cytoplasmic area of chromaffin cells of adrenal gland; and no. of endocrine cells/unit area and CSA of the Islets of Langerhans. The architectural distribution and histometery of Ns group did not differ much with that of Vc group. However in BfNs2.5 and BfNs5 the pace of rehabilitation of histoarchitecture of endocrine glands were much higher than that of Bf2.5 and Bf5 groups, clearly indicating the significance of Ns seed oil in rehabilitation of histological land marks of intoxicated mice endocrine glands.The Nigella sativa seed oil post treated group has shown rehabilitative effects against histopathologies. On the basis of these findings, it is concluded that insecticide harbor endocrine gland histopathological potentials that are at least partially rescue-able by Nigella sativa oil treatment. These findings thus indicate effective neutraceutical potential of Ns oil for possible human consumption in similar accidental exposure to the insecticide.