Histopathological and Immunohistochemical Study of the Effect of Sildenafil Citrate, Vitamin A, Vitamin C and Vitamin E on Wound Healing in Alloxan-induced Diabetic Rats

ABSTRACT Objective: To examine the impact of sildenafil citrate, vitamin A (Vit A), vitamin C (Vit C) and vitamin E (Vit E) on wound healing in a rat model of diabetes (n = 100). Methods: Diabetes was induced in the rats by intraperitoneal (i.p.) administration of alloxan. After anaesthesia, a standard wound was created on the back of each rat using a 10-mm sterile punch. The rats were randomly divided into 10 groups (n = 10 in each), as follows: normal saline, glibenclamide, insulin, sildenafil, Vit A, Vit C, Vit E, Vit A + sildenafil, Vit C + sildenafil, Vit E + sildenafil daily for 15 days. The rats were sacrificed after being anaesthetized 3, 7 and 15 days later. Wounded skin tissue samples were collected for histopathological and immunohistochemical analyses. Results: On the 7th day, epithelial regeneration was completed in groups 8 and 9. Angiogenesis was insufficient in group 2. In terms of connective tissue proliferation, partially matured connective tissue was observed in group 4. On the 15th day of the study, groups 8, 9 and 10 had mature connective tissue. However, group 1 still had exudate-containing neutrophils. Immunohistochemically, on the 3rd day, the level of inducible nitric oxide synthase (iNOS) reactivity was intense in macrophages and neutrophils surrounding the wound edges in groups 4, 8, 9 and 10. The level of iNOS reactivity was moderate in group 6 and less distinct in groups 1, 2, 3, 5 and 7. Conclusion: Sildenafil citrate, together with Vit A and Vit C, is beneficial in wound healing of diabetic rats.

Immunohistopathological and Biochemical Study of the Effects of Dead Nettle (Urtica Dioica) Extract on Preventing Liver Lesions Induced by Experimental Aflatoxicosis in Rats / Estudio inmunohistopatológico y bioquímico de los efectos del extracto de la ortiga mayor (Urtica dioica) en la prevención de lesiones hepáticas inducidas por aflatoxicosis experimental en ratas

ABSTRACT Objective: Aflatoxicosis is a mycotoxicosis infection with an acute or chronic course that forms due to aflatoxins (AFs) in humans and animals. Aflatoxins primarily affect the liver and can lead to histopathological necrosis, fibrosis and hepatocarcinogenesis of the organ. This paper studied the preventive effects of dead nettle leaf (Urtica dioica leaf; UDL) extract on liver lesions that were induced by experimental aflatoxicosis in rats. Methods: A total of 30 rats were separated into three groups of 10 rats each. Experimental group A (control) received normal rat food, experimental group B (AFB1) received 2 mg/kg of AF, and experimental group C (AFB1 + UDL extract) received 2 mg/kg of AF + 2 ml/rat/day of UDL extract. After three months of experimentation, blood and tissue samples were taken from the rats by necropsy to perform chemical and histopathological analyses. Results: According to the biochemical and histopathological findings, antioxidant system activity increased and lipid peroxidation and liver enzyme levels decreased in the group that received UDL extract. Conclusion: The extract of UDL had hepatoprotective effects against aflatoxicosis.

RESUMEN Objetivo: La aflatoxicosis es una infección por micotoxicosis con un curso agudo o crónico producido por aflatoxinas (AF) en seres humanos y animales. Las aflatoxinas afectan principalmente el hígado y pueden conducir a necrosis histopatológica, fibrosis o hepatocarcinogénesis del órgano. En este trabajo se estudiaron los efectos preventivos del extracto de la hoja de ortiga mayor (Urtica dioica l; UDL) sobre las lesiones hepáticas inducidas por aflatoxicosis experimental en ratas. Métodos: Un total de 30 ratas se separaron en tres grupos de 10 ratas cada una. EL grupo experimental A (control) recibió comida normal de ratas; el grupo experimental B (AFB1) recibió 2 mg/kg de AF; y el grupo experimental C (AFB1 + extracto de UDL) recibió 2 mg/kg de AF + 2 ml/rata/día de extracto de UDL. Después de tres meses de experimentación, se tomaron muestras de sangre y tejidos de las ratas en una necropsia encaminada a realizar análisis químicos e histopatológicos. Resultados: Según los hallazgos bioquímicos e histopatológicos, la actividad del sistema antioxidante aumentó, y la peroxidación del lípido y los niveles de la enzima del hígado disminuyeron en el grupo que recibió el extracto de UDL. Conclusión: El extracto de UDL tuvo efectos hepatoprotectores contra la aflatoxicosis.

Protective effects of Urtica dioica seed extract in aflatoxicosis: histopathological and biochemical findings.

The ameliorative potential and antioxidant capacity of an extract of Urtica dioica seeds (UDS) was investigated using histopathological changes in liver and kidney, measuring serum marker enzymes, antioxidant defence systems and lipid peroxidation (malondialdehyde (MDA)) content in various tissues of broilers exposed to aflatoxin (AF). A total of 32 broilers were divided randomly into 4 groups: control, UDS extract-treated, AF-treated and AF+UDS extract-treated. Broilers in control and UDS extract-treated groups were fed on a diet without AF. The AF-treated group and AF+UDS extract-treated groups were treated with an estimated 1 mg total AF/kg feed. The AF+UDS extract groups received in addition 30 ml UDS extract/kg diet for 21 d. The AF-treated group had significantly decreased body weight gain when compared to the other groups. Biochemical analysis showed a small increase in the concentrations of serum aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase and lactate dehydrogenase in the AF-treated group compared to that of the control group, whereas concentrations of these enzymes were decreased in the AF+UDS group compared to that of the AF-treated group. Administration of supplementary UDS extract helped restore the AF-induced increase in MDA and reduced the antioxidant system towards normality, particularly in the liver, brain, kidney and heart. Hepatorenal protection by UDS extracts was further supported by the almost normal histology in AF+UDS extract-treated group as compared to the degenerative changes in the AF-treated broilers. It was concluded that UDS extract has a protective hepatorenal effect in broilers affected by aflatoxicosis, probably acting by promoting the antioxidative defence systems.

Pulmonary myxoma in a sheep.

Pulmonary myxoma is an uncommon neoplasm. A pale tan, lobulated, and well-circumscribed mass was discovered at slaughter in the left lung of a 5-year-old sheep. Histologically, the tumor was composed of spindloid to stellate cells in a myxoid matrix. Neoplastic cells were immunohistochemically positive for vimentin but did not express cytokeratins, S-100 protein, smooth muscle actin, desmin, or p53. On the basis of the histologic and immunohistochemical findings, this tumor was diagnosed as a myxoma.

Immunohistochemical detection of Mannheimia (Pasteurella) haemolytica antigens in goats with natural pneumonia.

Pneumonia is a leading cause of loss to ruminants throughout the world. Mannheimia (Pasteurella) haemolytica is one of the most important etiological agent of pneumonia in cattle, sheep, and goats. This study was carried out to determine the incidence of M.haemolytica antigens using immunohistochemistry labelling of formalin-fixed, paraffin-embedded tissues in pneumonic lungs of goats slaughtered at abattoir, and then to compare these immunohistochemistry results with the results of bacterial isolation. For these objectives, a total of 1505 goat lungs slaughtered in slaughterhouse were grossly examined and pneumonia was detected in 74 cases (4.91%). Of these, with the exception of verminous pneumonia observed in 32 cases, on 42 pneumonic lungs immunohistochemical examinations were performed. Formalin-fixed and paraffin-embedded lung tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex (ABC) procedure using polyclonal antibodies to detect M.haemolytica antigens. Pneumonic lesions were more frequently encountered in cranioventral lobes than caudal lobes, and characterized by irregular lobular foci of atelectasis or lobar pneumonia. The presence of M.haemolytica antigens was detected in 19 (45%) out of 42 pneumonic lungs. Bacterial antigens were found most frequently in the cytoplasm of bronchial and bronchiolar epithelial cells, in the swirling degenerating leukocytes in the alveoli, and in the degenerating leukocytes in the area of coagulation necrosis, less frequently in the epithelial cells of bronchial glands, and lymphoid cells. Conclusionly, immunohistochemical detection of M.haemolytica antigens in pneumonic lungs appear to be more reliable compared to bacterial isolation.

Comparison of fibrin glue and suture in the healing of teat incisions in lactating goats.

The aims of this study were to investigate whether fibrin glue can be used to close experimentally induced incisions of the teat (mammary papillae) in lactating goats and to compare the healing of the glued with the sutured incisions. Four clinically healthy lactating dairy goats, namely 8 mammary papillae were used. After surgical preparation of the papillae, a 3.5 cm long incision of each papilla was made through skin, muscular layer and mucosa into the papillary sinus. The wounds in the right papillae in all goats were closed with U-shaped uninterrupted 00 chromic catgut sutures. The wounds in the left papillae in all goats were closed, using fibrin glue. One incision was seen to be dehisced and fistulous one day after in fibrin glued teats. The animals were slaughtered 8 days after surgical manipulation. The mammary papillae were removed and examined in the viewpoint on gross and microscopic findings. The healing of wounds was slower and feeble in glued mammary papillary incisions, however faster and stronger in sutured incisions on day 8 after operations. But, available outcomes like less tissue thickness and positive cosmetic results could be obtained byfibrin glue used on mammary papillary incisions, which are very important for teats to be milked by hand and milking machine. Results suggest that it is advisable to use only one or two simple interrupted sutures in teat incisions glued with fibrin to prevent the dehiscence but with a more reliable healing than the sutured incisions.

Immunohistochemical detection of parainfluenza type 3 virus antigens in paraffin sections of pneumonic caprine lungs.

Pneumonia is a leading cause of loss to ruminants throughout the world. Parainfluenza type-3 virus (PI-3) is one of the most important respiratory pathogens of bovine and ovine. In this study, prevalence of PI-3 virus infection as causative agent of pneumonia in goats was investigated. For this purpose, a total of 1505 goat lungs slaughtered in Bitlis and Van slaughterhouses were grossly examined and pneumonia was detected in 74 cases (4.91%). Lesions were more frequently encountered in anteroventral lobes than caudal lobes. With the exception of verminous pneumonia observed in 32 cases, immunohistochemical examinations were performed on 42 pneumonic lungs. Formalin-fixed and paraffin-embedded lung tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex procedure using polyclonal antibodies to detect PI-3 viral antigens. The presence of PI-3 viral antigens was detected in 28 (66.6%) of 42 pneumonic lungs. Viral antigens were found most frequently in the cytoplasm of bronchiolar epithelial cells, type II pneumocytes, and less frequently in the epithelial cells of bronchial glands, syncytial cells, alveolar macrophages, and lymphocytes and plasma cells. In conclusion, it was found that there was a close relationship between the pneumonia in goats and the presence of PI-3 viral antigens. Incidence of PI-3 virus in pneumonic lungs of goats was detected to be very high in the present study performed in the region of Bitlis and Van, Turkey.

The role of Urtica dioica and Nigella sativa in the prevention of carbon tetrachloride-induced hepatotoxicity in rats.

The role of Nigella sativa L. (Ranunculaceae) (NS) and Urtica dioica L. was investigated (UD) in the prevention of carbon tetrachloride (CCl4) induced liver fibrosis and cirrhosis. Fifty Sprague-Dawley rats were allocated into fi ve groups (I, IIA and B, IIIA and B) and CCl4 was injected biweekly to all groups. Group I (control, CCl4 only), group IIA and B (NS fixed oil and volatile oil), group IIIA and B (UD fixed oil and UD decoction extract) rats were killed at the end of week 12 and histopathological and immunohistochemical examinations of liver tissues were performed. In the control group, coagulation necrosis and hydropic degeneration were marked in the periacinar regions (zone 3) associated with fibrosis in the periacinar regions and in the portal tracts. In groups IIA-B and IIIA-B (NS and UD), none of the serious histopathological findings were detected except for sparse coagulation necrosis in the periacinar regions. ASMA-positive perisinusoidal cells with myo fibroblastic transformation and lysosomal enzyme activity suggesting fibrogenesis were also significantly more common in the control group than in the NS and UD groups. UD and NS seem to be significantly effective in the prevention of carbon tetrachloride induced hepatotoxicity in rats.

Undifferentiated ganglioneuroblastoma in a sheep.

Ganglioneuroblastoma, presumably originating from the adrenal medulla, was found in an 18-month-old sheep. Histologically, the tumour was mainly composed of undifferentiated neuroblasts with neuropil and occasional ganglion cells. The morphological and immunohistochemical features with respect to intermediate filament proteins and the neuroendocrine nature of the tumour are described. The neoplastic cells were positive for neuron-specific enolase, synaptophysin and vimentin.

Immunoperoxidase and histopathological examinations of leptospiral nephritis in cattle.

This study was designed to investigate the role of leptospirae in interstitial nephritis. Sixty-eight white-spotted kidneys and 30 grossly normal kidneys from slaughtered cattle were examined histologically and immunohistochemically for the presence of Leptospira interrogans antigens. The presence of L. interrogans antigens was found in 21 of 68 white-spotted kidneys and in four of 30 normal kidneys. In conclusion, the detected incidence of infection with L. interrogans was not high, but there was a relationship between the presence of interstitial nephritis and leptospiral antigens.

Alveolar rhabdomyosarcoma of the limb in a sheep.

Tumor mass was detected in the skeletal musculature of the tibia in a sheep slaughtered at abattoir. Histologically, the tumor cells were in a typical alveolar pattern, and showed a cartilaginous tissue differentiation in some areas of the tumor mass. This tumor examined by avidin-biotin-peroxidase method was found to be positive for myoglobin. The diagnosis of undifferentiated alveolar rhabdomyosarcoma was based on the results of histopathology and immunohistochemistry.

The role of antioxidant vitamins (C and E), selenium and Nigella sativa in the prevention of liver fibrosis and cirrhosis in rabbits: new hopes.

This experiment was carried out to investigate the role of antioxidants such as vitamin C and E, selenium and Nigella sativa (NS) on the prevention of carbon tetrachloride (CCl4)-induced liver fibrosis in rabbits. It was found that superoxide dismutase (SOD) values in all of the treated groups were significantly lower than those of the control at 12th week of experiment (p < 0.05), while at 6th week and 12th week of experiment glutathione peroxidase (GSH-Px) values in the vitamin C treated group were significantly different from the control (p < 0.05). Histopathologically, hepatocellular necrosis, degeneration and advanced fibrosis were found in the control group. Lesions were minor and only confined to midzonal regions without centrilobular necrosis and fibrosis in the NS treated animals (group B). The lesions observed in the vitamin C treated animals (group C) were similar to that of the control group. Parenchymal changes with fibrosis were less in selenium and vitamin E treated animals (group D) than in those of the control group, but more obvious than in NS group. Histopathological findings demonstrate that NS might, at least partly, be successful in the prevention of liver fibrosis in rabbits. Vitamin E plus selenium had little therapeutic effect and vitamin C seemed to be ineffective, as far as the results of this study are concerned.

Effect of Nigella sativa on glucose concentration, lipid peroxidation, anti-oxidant defence system and liver damage in experimentally-induced diabetic rabbits.

This study was carried out to investigate whether Nigella sativa could decrease the lipid peroxidation, increase the anti-oxidant defence system and also prevent the lipid-peroxidation-induced liver damage in experimentally induced diabetic rabbits. Fifteen New Zealand male rabbits were divided into three experimental groups: control, diabetic and diabetic and N. sativa-treated. The diabetes mellitus (DMI) was induced in the rabbits using 150 mg/kg of 10% alloxan. The diabetic + N. sativa-treated group was given extract of N. sativa seeds orally every day for 2 months after induction of DM. At the end of the 2-month experiment, blood samples were collected to measure malondialdehyde (MDA), glutathione (GSH), ceruloplasmin and glucose concentration, and livers were harvested for histopathological analysis. Treatment with N. sativa decreased the elevated glucose and MDA concentrations, increased the lowered GSH and ceruloplasmin concentrations, and prevented lipid-peroxidation-induced liver damage in diabetic rabbits. It was concluded that N. sativa might be used in diabetic patients to prevent lipid peroxidation, increase anti-oxidant defence system activity and also prevent liver damage.