Crab shell extract improves serum biochemical markers and histological changes of pancreas in diabetic rats / El extracto de cáscara de cangrejo mejora los marcadores bioquímicos del suero y los cambios histológicos del páncreas en ratas diabética

SUMMARY: Diabetes mellitus is a common metabolic disease. There are many natural agents available to control and treat diabetes. Crab shell extract has antioxidant properties. The aim of present study was to investigate the effect of crab shell hydroalcoholic extract on blood glucose, liver enzymes, nitric oxide and antioxidant capacity of serum and histological structure of pancreas in diabetic rats. In this experimental study, thirty five male Wistar rats (180-220 g) were divided into control, diabetic and experimental groups (n=7). Diabetes was induced by intraperitoneal injection of streptozotocin (60 mg/kg). Rats were treated for 14 days by crab shell extract with 100, 200 and 400 mg/kg doses. Fasting blood glucose, serum levels of liver enzymes, nitric oxide (NO) and total antioxidant capacity were evaluated. Changes of pancreatic tissue were determined using a modified aldehyde fuchsin staining method. Data were analyzed using one-way ANOVA. Differences were considered statistically significant at P<0.05. Crab shell extract induced a significant reduction in blood glucose, serum levels of nitric oxide and ALT (P=0.033). Also, there were a significant increase in total antioxidant capacity (FRAP) (P=0.007), and insignificant decrease in serum levels of AST. The extract improved pancreatic tissue changes caused by diabetes. In conclusion, antioxidant and anti-diabetic effects of crab shell increase total antioxidant capacity of serum and decreased blood glucose, serum nitric oxide and ALT levels.

RESUMEN: La diabetes mellitus es una enfermedad metabólica común. Hay muchos agentes naturales disponibles para controlar y tratar la diabetes. El extracto de cáscara de cangrejo tiene propiedades antioxidantes. El objetivo del presente estudio fue investigar el efecto del extracto hidroalcohólico de la cáscara de cangrejo sobre la glucosa sérica, las enzimas hepáticas, el óxido nítrico y la capacidad antioxidante del suero y la estructura histológica del páncreas en ratas diabéticas. En este estudio experimental, treinta y cinco ratas Wistar machos (180220 g) se dividieron en cinco grupos: control, diabéticos y experimentales (n = 7). La diabetes se indujo por inyección intraperitoneal de estreptozotocina (60 mg / kg). Las ratas se trataron durante 14 días con extracto de cáscara de cangrejo con dosis de 100, 200 y 400 mg / kg. Se evaluaron la glucosa en sangre en ayunas, las enzimas hepáticas, el óxido nítrico sérico y la capacidad antioxidante total. Los cambios en el tejido pancreático se determinaron usando un método de tinción de aldehído fucsina modificado. Los datos se analizaron utilizando ANOVA unidireccional. Las diferencias se consideraron estadísticamente significativas a P <0,05. El extracto de cáscara de cangrejo indujo una reducción significativa en la glucosa en sangre, en los niveles séricos de óxido nítrico y ALT (P = 0,033). Además se observó un aumento significativo en la capacidad antioxidante total (FRAP) (P = 0.007), y una disminución insignificante en los niveles séricos de AST. El extracto mejoró los cambios en el tejido pancreático causados por la diabetes. En conclusión, los efectos antioxidantes y antidiabéticos de la cáscara de cangrejo aumentan la capacidad antioxidante total de suero y la disminución de la glucosa en la sangre, el óxido nítrico sérico y los niveles de ALT.

Adiponectin effect on nitric oxide secretion by normal and endometriotic human endometrial stromal cells: in vitro study / Efecto de la adiponectica sobre la secreción de óxido nítrico por células estromales de endometrio humano, normales y con endometriosis: estudio in vitro

Endometriosis is an estrogen-dependent disease in reproductive age women. Adiponectin and Nitric oxide (NO) have an important role in physiologic functions especially in human reproductive system. Levels of NO increased in the endometriosis patients but serum adiponectin levels decreased in woman with endometriosis. The aim of this study was to determine adiponectin effect on nitric oxide secretion by cultured normal and endometriotic human endometrial stromal cells. In this experimental study, normal (n= 10) and endometriotic endometrial biopsies (n= 10) were taken in sterile condition. Stromal cells isolated and cultured in in DMEM/ F12 medium and treated with adiponectin concentrations (0, 10, 100, and 200 ng/ml) for 24 and 48 hours. NO assay was done on their supernatants by Greiss method. Data was analyzed by one way ANOVA and p<0.05 was considered significant. There was significant difference between endometriosis groups in NO secretion in all dose of adiponectin and time (p<0.05). In normal groups there was significant difference in 48 hours (p<0.05) but no significant change in 24 hours (p>0.05). Adiponectin effects nitric oxide secretion of cultured human endometriotic stromal cells.

La endometriosis es una enfermedad dependiente de estrógenos que se presenta en mujeres en edad reproductiva. La adiponectina y el óxido nítrico (ON) tienen un papel importante en las funciones fisiológicas, especialmente en el sistema reproductivo humano. Los niveles de ON aumentan en los pacientes con endometriosis, pero los niveles de adiponectina en suero disminuyen. El objetivo fue determinar el efecto de la adiponectina sobre la secreción de ON por las células estromales de endometrio humano, tanto normales como con endometriosis, en medio de cultivo. En este estudio experimental, las células estromales de endometrio normales (n= 10) y las biopsias de endometrio con endometriosis (n= 10) se tomaron en condiciones de esterilidad. Las células estromales fueron aisladas y cultivadas en un medio DMEM/F12, y se sometieron a distintas concentraciones de adiponectina (0, 10, 100, y 200 ng/ml) durante 24 y 48 horas. El ensayo con ON se realizó a los sobrenadantes obtenidos por el método de Greiss. Los datos recolectados fueron analizados por ANOVA de una vía y un valor p<0,05 se consideró significativo. Entre los grupos con endometriosis, en referencia a la secreción de ON, no hubo diferencia significativa en todas las dosis de adiponectina y los tiempos estipulados (p<0,05). En los grupos normales, hubo diferencia significativa a las 48 horas (p<0,05), pero ningún cambio significativo a las 24 horas (p>0,05). La adiponectina tiene efectos sobre la secreción de óxido nítrico por las células estromales endometriales humanas en cultivo.

Neuronal differentiation of PC12 and embryonic stem cells in two- and three-dimensional in vitro culture.

The quality of neuronal differentiation and reduction in apoptosis that occurred in two-dimensional (2D) and three-dimensional (3D) culture conditions is compared. PC12 and embryonic stem cells are two commonly utilized cell lines for the study of neuronal regeneration. These cells were induced to neuronally differentiate by adding NGF and retinoic acid respectively. Total neurite length and expression of neuronal markers (MAP-2 and β3-tubulin) was assessed by morphometry and immunocytochemistry. Also, TUNEL assay was used to detect apoptosis. Upon exposure to a differentiation media in the 3D fibrin gel, PC12 and embryonic stem cells stopped dividing, had increased adhesion to the substratum, extended neurite processes and expressed neuronal markers. The same results, however, were not observed with the 2D culture. Also, the apoptosis index performed by TUNEL assay demonstrated a reduction in the degree of apoptosis in the 3D culture compared to 2D culture. Fibrin matrix supports growth and neuronal differentiation of PC12 and embryonic stem cells. In addition, the 3D culture enhanced cellular resistance to apoptosis when compared to the 2D culture. It appears as if a 3D culture system may offer a better technique for future neuronal tissue engineering investigations.