Effect of Thymoquinone on skeletal muscle regeneration via assessment of Pax-7 and Myo-D expression in the DMBA-treated hamster pouch / Efeito da Timoquinona na regeneração muscular esquelética através da avaliação da expressão das proteínas Pax-7 e Myo-D em bolsa jugal de hamsters tratados com DMBA

Objective: Pax-7 and Myo-D regulate satellite cells' activation and differentiation, thus muscle regeneration following damage. This research aimed to investigate the effect of Thymoquinone (TQ) on skeletal muscle regeneration following 7,12-dimethylbenz-(a)-anthracene (DMBA)-induced injury in the hamster buccal pouch via immunohistochemical assessment of Pax-7 and Myo-D expression. Material and Methods: 65 male golden Syrian hamsters were divided into 3 groups: Group 1: (n=5) received no treatment. Group 2: (n=20) served as a positive control. The left buccal pouches were painted with the carcinogen 3/week/ 6weeks. Group 3: (n=40) were subdivided into two equal sub-groups as follows: Group 3a: (n=20) were given one i.p. TQ injection. Group 3b: (n=20) were given two i.p. TQ injections. Five animals from each group (2 and 3) were euthanized at 24, 48 hrs, one, and two weeks after the last injection. A blood sample (2 ml) was withdrawn for assessment of TNF-α levels in serum. Serial sections of the pouches were examined histologically (H&E), and immunohistochemically (IHC) for the detection of Pax-7 and Myo-D proteins. Results: double i.p injections of TQ resulted in a significant elevation in the level of TNF-α from the second-day post-injection with a progressive formation of the muscle fibers (MFs) and mononuclear cells (MNCs) around the deeper blood vessels. At 14 days, no statistically significant difference was found between this group and group '2', while the difference remained significant compared to groups '1' and '3a'. The muscle fibers were more mature and compact. IHC results showed positive expression of the perivascular mononuclear cells (MNCs) to both Pax-7 and Myo-D with positive reactivity of the peripheral nuclei of muscle fibers to Pax-7 compared to the negative reaction in the positive control group. Conclusion: early and two TQ injections had a promising effect on the induction of striated muscle regeneration, mainly by non-myogenic stem cells (AU)

Objetivo: Pax-7 e Myo-D regulam a ativação e diferenciação de células satélites durante a regeneração muscular pós-trauma. Assim, objetivamos investigar o efeito da timoquinona (TQ) na regeneração muscular esquelética após injúria causada por 7,12 dimetilbenzantraceno (DMBA) em bolsa jugal de hamsters, através da análise imuno-histoquímica de Pax-7 e Myo-D. Material e Métodos: 65 hamsters-sírios machos foram divididos em 3 grupos: Grupo 1: (n=5) controle negativo, sem tratamento. Grupo 2: (n=20) controle positivo. A bolsa jugal do lado esquerdo recebeu aplicação do DMBA por 3 e 6 semanas. Grupo 3: (n=40) receberam aplicação de DMBA e foram então subdivididos em: Grupo 3a: (n=20) que recebeu 1 injeção intraperitoneal (ip) de TQ e Grupo 3b: (n=20) que recebeu duas injeções ip de TQ. Cinco animais dos grupos 2 e 3 foram eutanasiados em 24 horas, 48 horas, 7 dias e 14 dias após a administração de DMBA e da última injeção de TQ. Amostras de sangue (2 ml) foram coletadas para avaliação dos níveis séricos de TNF-α. Cortes seriados da bolsa jugal dos animais foram analisados histologicamente (H&E), e através de imunohistoquimica (IHC) para avaliação das proteínas Pax-7 e Myo-D. Resultados: duas injeções ip de TQ aumentaram os níveis séricos TNF-α à partir do segundo dia pós-administração com formação progressiva de fibras musculares (MFs) e células mononucleares (MNCs) ao redor dos vasos sanguíneos. No dia 14, não houve diferença estatística entre o grupo 3b e o grupo 2, enquanto a diferença permaneceu entre o grupo 1 e 3a. As MFs apresentavam-se mais maduras e compactas. A IHC mostrou expressão de Pax-7 e Myo-D nas MNCs ao redor dos vasos, e houve expressão nuclear de Pax-7 nas MFs no grupo 2. Conclusão: ambos regimes de administração do TQ, 1 ou 2 aplicações ip, apresentaram efeito promissor na indução da regeneração muscular esquelética, principalmente nas células não-miogênicas.(AU)

Prevention of doxorubicin-induced experimental cardiotoxicity by Nigella sativa in rats.

INTRODUCTION: Doxorubicin (DOX) is an anthracycline cytotoxic chemotherapeutic drug that is commonly used in cancer treatment. A major side effect limiting the clinical use of DOX is cardiotoxicity due to oxidative injury. Nigella sativa (NS) is an annual flowering plant with antioxidant properties. Its seeds contain several bioactive constituents such as saturated and unsaturated fatty acids, thymoquinone, dithymoquinone, thymohydroquinone, and thymol. In this study, we investigated the effect of NS extract on DOX-induced cardiotoxicity. METHODS: The experimental study animals consisted of 28 male Sprague Dawley rats weighing between 300 and 400 g. Four study groups each of seven rats were defined: controls; NS extract; DOX; and DOX+NS. Control and DOX rats received standard food, while each rat in the NS and DOX+NS groups also received 100 mg/kg NS extract orally. At day 28 of follow-up, rats in the DOX groups were administered a single 10 mg/kg intraperitoneal dose of DOX, while rats in the control and NS groups received a single 10 mg/kg dose of physiological saline solution. All animals were monitored for 35 days. On day 35, the rats were decapitated and serum and cardiac tissue samples were obtained. Troponin and NT-proBNP levels were measured in blood sera, while malondialdehyde (MDA), nitric oxide, total antioxidant capacity (TAC), and total oxidative stress (TOS) levels were quantified in sera and tissue samples. Histological alterations that were assessed in cardiac tissue included myocyte disarray, small vessel disease, myocyte hypertrophy, and fibrosis. RESULTS: The DOX group had significantly higher NT-proBNP, TOS, and MDA, with greater histopathological derangement. TAC was significantly elevated in the DOX+NS group, which also exhibited significantly lower troponin, TOS, and MDA, as well as significantly higher TAC compared to the DOX group. Histopathological examination showed that the significant structural derangement observed in DOX rats was markedly and significantly reduced in DOX+NS rats. CONCLUSION: Our results suggest that NS extract may prevent DOX-induced cardiotoxicity and thus represents a promising cardioprotective agent.

Investigation of the protective and therapeutic effects of ß-aminoisobutyric acid (BAIBA) and Thymoquinone in the diabetic nephropathy / Investigación de los efectos protectores y terapéuticos del ácido ß-aminoisobutírico (BAIBA) y la timoquinona en la nefropatía diabética

In this study, against streptozotocin (STZ) induced diapetic nephropathy (DN); it is aimed to investigate the use of thymoquinone (TQ) and ß-aminoisobutyric acid (BAIBA) and to compare the effects of these agents. With random selection of 35 male rats, five groups (seven rats in each group) were constituted as follows: Control, STZ, STZ + TQ, STZ + BAIBA, STZ + TQ + BAIBA. In the STZ group; body weight, glutathione (GSH) and insulin levels decreased, relative kidney weight, malondialdehyde (MDA), glucose, blood urea nitrogen (BUN) and creatinine (Cr) levels were increased. Also, in kidney tissue; histopathological changes (such as thickening of the capsular, glomerular and tubular basement membranes, increased mesangial matrix amount, increased cytoplasmic vacuolization in some of the tubular epithelial cells, increased tumor necrosis factor-alpha (TNF-α) expression, and inflammatory cell infiltrations in interstitial tissue) were detected. It was observed that these changes occurring after diabetes mellitus (DM) reversed significantly in TQ, BAIBA and TQ + BAIBA groups.

En este estudio, contra la nefropatía diapética (ND) inducida por estreptozotocina (STZ); tiene como objetivo investigar el uso de timoquinona (TQ) y ácido ß-aminoisobutírico (BAIBA) y comparar los efectos de estos agentes. Con la selección aleatoria de 35 ratas macho, se constituyeron cinco grupos (siete ratas en cada grupo) como sigue: Control, STZ, STZ + TQ, STZ + BAIBA, STZ + TQ + BAIBA. En el grupo STZ; el peso corporal, los niveles de glutatión (GSH) y de insulina disminuyeron, el peso relativo de los riñones, el malondialdehído (MDA), la glucosa, el nitrógeno ureico en sangre (BUN) y los niveles de creatinina (Cr) aumentaron. Además, en tejido renal; se detectaron cambios histopatológicos (como engrosamiento de las membranas basales capsular, glomerular y tubular, aumento de la cantidad de matriz mesangial, aumento de la vacuolización citoplasmática en algunas de las células epiteliales tubulares, aumento de la expresión del factor de necrosis tumoral alfa (TNF-α) e infiltraciones de células inflamatorias en tejido intersticial). Se observó que estos cambios que ocurren después de la diabetes mellitus (DM) se revirtieron significativamente en los grupos TQ, BAIBA y TQ + BAIBA.

Thymoquinone ameliorates acute kidney injury induced by renal ischemia-reperfusion / La timoquinona mejora la lesión renal aguda inducida por isquemia-reperfusión renal

SUMMARY: Renal ischemia-reperfusion injury (IRI)is an unavoidable consequence in renal transplantation and multiple clinical settings. A debate has been raised about the particular role of hypoxia-inducible factor (HF-1α) in the renal injury pathogenesis and the renal cortex ultrastructural alterations. Also, we investigated the antioxidant/anti-inflammatory effect of thymoquinone and its modulatory role on HIF-1α in protection against renal IRI.Adult male Wister albino rats were assigned into 3 groups (n=16); 1) Sham-operated, 2) IRI model and 3) renal IRI pre-treated with thymoquinone 10 mg.kg-1.day-1 (TQ-IRI) for 10 days and at the reperfusion onset. Following the operation, 8 rats from each group were euthanized after 3 hours and the remaining 8 rats at 24 hours. Renal injury was assessed by the increased blood urea nitrogen, creatinine level, and the EGTI histological injury scoreat both 3 and 24h. HIF-1α was upregulated (p<0.01) and was correlated with renal tissue reactive oxygen species (ROS) production and total oxidant capacity (TAC) consumption. Elevated inflammatory markers (NFkB, MCP-1 and VCAM-1) were associated with renal IRI.Thymoquinone treatment inhibited the accumulation of HIF-1α (p<0.01), reduced renal oxidation/inflammation process and markedly diminished renal injury.

RESUMEN: La lesión por isquemia-reperfusión renal (IRR) es una consecuencia inevitable en el trasplante renal como también en múltiples contextos clínicos. Se ha suscitado una discusión referente a la relación particular del factor inducible por hipoxia (HF- 1α) en la patogénesis de la lesión renal y las alteraciones ultraestructurales de la corteza renal. Además, investigamos el efecto antioxidante / antiinflamatorio de la timoquinona y su papel modulador sobre HIF-1α en la protección contra IRR. Se utilizaron ratas albinas Wister macho adultas divididas en 3 grupos (n = 16); 1) Intervención simulada, 2) modelo IRR y 3) IRR pretratado con timoquinona 10 mg/kg-1. día-1 (TQ-IRR) durante 10 días y al inicio de la reperfusión. Posterior a la operación, 8 ratas de cada grupo fueron sacrificadas después de 3 horas y las 8 ratas restantes a las 24 horas. La lesión renal se evaluó por el aumento de nitrógeno ureico en sangre, el nivel de creatinina y la puntuación de lesión histológica EGTI tanto a las 3 como a las 24 horas. HIF-1α se incrementó (p <0,01) y se correlacionó con la producción de especies de oxígeno reactivo (ROS) del tejido renal y el consumo de capacidad oxidante total. Los marcadores inflamatorios elevados (NFkB, MCP-1 y VCAM-1) se asociaron con IRR. El tratamiento con timoquinona inhibió la acumulación de HIF-1α (p <0,01), redujo el proceso de oxidación / inflamación renal y disminuyó notablemente la lesión renal.

Therapeutic effect of thymoquinone against methotrexate- induced damage on sperm parameters in mice / Efecto terapéutico de la tiroyoquinona contra el daño inducido por metotrexato en los parámetros espermáticos en ratones

Methotrexate drug is commonly used to treat cancer; it is known to cause reproductive damage. Thymoquinone, as a natural component of herbs has many healthy benefits shown in researches. The present study aimed to investigate probable therapeutic effects of Thymoquinone against Methotrexate-induced damage on sperm parameters in mice. In this experimental study, 30 male mice (25-30 g) were divided into five groups of six in each group. The mice were received normal saline (control group), Methotrexate (20 mg/kg), Methotrexate (20 mg/kg) + Thymoquinone (2, 10 and 20 mg/kg) by intraperitoneal injection. On the day after the last injection, the sperm parameters including motility, viability and count of sperms were assessed. Data analysis was performed using one-way ANOVA followed by Tukey test. Methotrexate alone showed a significant reduction in sperm parameters compared to the control group (P=0.00). In groups treated with Methotrexate and Thymoquinone, sperm parameters (motility ,viability, count sperm) did not show any significant differences with control group (P=0.00). Thymoquinone, as a potent antioxidant, could compensate for the toxicity induced by Methotrexate. These medical trends may be useful for diminishing the side effects of Methotrexate on the male reproductive system.

El metotrexato es un fármaco utilizado comúnmente para tratar el cáncer pero además causa daño en los órganos reproductivos. Durante las investigaciones se ha demostrado que la timoquinona, un componente natural de las hierbas, tiene numerosos beneficios. El objetivo del estudio fue investigar el probable efecto terapéutico de la timoquinona contra el daño inducido por metotrexato, en los parámetros espermáticos en ratones. En este estudio experimental, se dividieron 30 ratones machos (25-30 g) en cinco grupos de seis en cada uno. Los ratones recibieron solución salina normal (grupo control), metotrexato (20 mg / kg), metotrexato (20 mg / kg) + timoquinona (2, 10 y 20 mg / kg) por inyección intraperitoneal. El día después de la última inyección, se evaluaron los parámetros espermáticos, incluida la motilidad, la viabilidad y el recuento de espermatozoides. El análisis de los datos se realizó utilizando test de ANOVA seguido de la prueba de Tukey. Durante el uso exclusivo de metotrexato se observó una reducción significativa en los parámetros espermáticos en comparación con el grupo control (P = 0.00). En los grupos tratados con metotrexato y timoquinona, los parámetros espermáticos (motilidad, viabilidad, conteo de espermatozoides) no mostraron diferencias significativas con el grupo control (P = 0.00). Como potente antioxidante, la timoquinona podría compensar la toxicidad inducida por metotrexato. Estas tendencias médicas pueden ser útiles para disminuir los efectos secundarios de metotrexato en el sistema reproductivo masculino.