Involvement of SIRT1 and SIRT3 in Effects of Hypoxic Hypobaric Preconditioning on Hippocampal Damage Induced by Chronic Cerebral Hypoperfusion in C57/6j and a7nAchRs(-/-) Mice / Participación de SIRT1 y SIRT3 en los Efectos del Precondicionamiento Hipobárico Hipóxico sobre el Daño del Hipocampo Inducido por Hipoperfusión Cerebral Crónica en Ratones C57/6j y a7nAchRs (-/-)

SUMMARY: Hypoxic preconditioning is known to induce neuroprotection, but its effects and pathways in chronic brain pathology still unknown. The aim was to establish an involvement of a7 subunit of nicotinic acetylcholine receptors (a7nAchRs), and sirtuins of 1 (SIRT1) and 3 (SIRT3) types in the effects of hypoxic hypobaric preconditioning on brain damage in mice with chronic cerebral hypoperfusion caused by the left common carotid artery occlusion. The male C57/6j (C57, wild type) and a7nAchRs(-/-) mice were divided to six experimental groups (10 mice per group): sham-operated C57, C57 with chronic cerebral hypoperfusion, C57 with hypoxic hypobaric preconditioning and chronic cerebral hypoperfusion, sham-operated a7nAchRs(-/-) mice, a7nAchRs(-/-) with chronic cerebral hypoperfusion, a7nAchRs(-/-) with hypoxic hypobaric preconditioning and chronic cerebral hypoperfusion. For preconditioning, mice were exposed to hypoxia by "lifting" in barochamber to simulated altitude of 5600 m a.s.l. for 1 h/day on 3 consecutive days before surgical manipulation. Expressions of SIRT1, SIRT3 in brain tissue, and histopathological changes of the hippocampi were examined. It was shown that 8-week chronic hypoperfusion of the brain, caused by unilateral occlusion of the common carotid artery, was accompanied by injury to the neurons of the hippocampi of both hemispheres, which was more pronounced on the side of the occlusion. This damage, as well as the mechanisms of neuroprotection induced by hypoxic preconditioning, were maintained for at least 8 weeks by mechanisms mediated through a7nAChRs. Deficite of a7nAChRs was accompanied with reduction of neuronal damage caused CCH in 8 weeks, as well as preconditioning effects, and lead to compensatory activation of regulatory and protective mechanisms mediated by SIRT1, in normal conditions and in CCH. In wild-type (C57) mice, protective mechanisms in CCH were realized to a greater extent by increased expression of SIRT3 in both hemispheres of the brain.

Se sabe que el precondicionamiento hipóxico induce neuroprotección, pero aún se desconocen sus efectos y vías en la patología cerebral crónica. El objetivo fue establecer la participación de la subunidad a7 de los receptores nicotínicos de acetilcolina (a7nAchR) y las sirtuinas de tipo 1 (SIRT1) y 3 (SIRT3) en los efectos del precondicionamiento hipóxico hipobárico sobre el daño cerebral en ratones con hipoperfusión cerebral crónica causada por la oclusión de la arteria carótida común izquierda. Los ratones macho C57/6j (C57, tipo salvaje) y a7nAchRs(-/-) se dividieron en seis grupos experimentales (10 ratones por grupo): C57 con operación simulada, C57 con hipoperfusión cerebral crónica, C57 con precondicionamiento hipobárico hipóxico y crónica. hipoperfusión cerebral, ratones a7nAchRs(-/-) operados de forma simulada, a7nAchRs(-/-) con hipoperfusión cerebral crónica, a7nAchRs(-/-) con precondicionamiento hipobárico hipóxico e hipoperfusión cerebral crónica. Para el preacondicionamiento, los ratones fueron expuestos a hipoxia "levantándolos" en una cámara de barro a una altitud simulada de 5600 m s.n.m. durante 1 h/día durante 3 días consecutivos antes de la manipulación quirúrgica. Se examinaron las expresiones de SIRT1, SIRT3 en tejido cerebral y los cambios histopatológicos de los hipocampos. Se demostró que la hipoperfusión cerebral crónica de 8 semanas, causada por la oclusión unilateral de la arteria carótida común, se acompañaba de lesión de las neuronas del hipocampo de ambos hemisferios y que era más pronunciada en el lado de la oclusión. Este daño, así como los mecanismos de neuroprotección inducidos por el precondicionamiento hipóxico, se mantuvieron durante al menos 8 semanas mediante mecanismos mediados por a7nAChR. El déficit de a7nAChR se acompañó de una reducción del daño neuronal causado por CCH en 8 semanas, así como de efectos de precondicionamiento, y condujo a una activación compensatoria de mecanismos reguladores y protectores mediados por SIRT1, en condiciones normales y en CCH. En ratones de tipo salvaje (C57), los mecanismos de protección en CCH se realizaron en mayor medida mediante una mayor expresión de SIRT3 en ambos hemisfe- rios del cerebro.

Anticonvulsant effects of Paeonia daurica subsp. macrophylla root extracts in pentylenetetrazol-induced seizure models in mice.

INTRODUCTION: In the present study, anticonvulsant effects of aqueous extract (AE), hydro-alcoholic crude extract (HE), and its fractions (F-CHCl3, F-EtOAc, F-MeOH) of Paeonia daurica subsp. macrophylla (P. daurica ssp. macrophylla) root examined by using a pentylenetetrazol-induced model (PTZ) on mice. METHODS: HE and its fractions as well as AE, in concentrations of (100, 200 and 400mg/kg), valproate (Val) (100 and 200mg/kg), and saline (negative control) (10mg/kg) were injected intraperitoneally (i.p.) 30min before PTZ (80mg/kg, i.p.). The time taken before the onset of myoclonic convulsions (MC), MC duration, time taken before the onset of generalized tonic-clonic seizures (GTCS), the duration of GTCS, and the percentage of GTCS and mortality protection recorded. The plant's anticonvulsant mechanisms were assessed using flumazenil (5mg/kg, i.p.) before AE (100, 200, and 400mg/kg, i.p.) injection. GraphPad Prism software was used to compare the differences between various treatment groups with one-way analysis of variance (ANOVA) followed by Tukey-Krammer multiple comparison tests. RESULTS: All the plant samples except F-EtOAc significantly delayed the onset and decreased the duration of PTZ-induced MCS and GTCS, and significantly reduced the GTCS and mortality rate. Pretreatment with flumazenil diminished the significant anticonvulsant effects of AE against PTZ-induced seizures. CONCLUSIONS: It can report that extract of P. daurica ssp. macrophylla might be a helpful guide for future studies in the treatment of epilepsy.

Assessing cell chimerism in an acute graft-versus-host disease model established in TLR4 knockout mice.

BACKGROUND: Graft-versus-host disease (GVHD) is a major complication after allogeneic hematopoietic stem cell transplantation. OBJECTIVE: To elucidate the role of Toll-like receptor 4 (TLR4), the major receptor for bacterial lipopolysaccharide, in the development of GVHD, we constructed a GVHD model in TLR4 knockout (TLR4-/-) mice and monitored the cell chimerism. METHODS: In this study, we used polymerase chain reaction to identify whether TLR4 knockout (TLR4-/-) mice were established. Before transplantation, we pretreated mice with irradiation so as to obtain an appropriate irradiation dose. Flow cytometry was applied to measure the chimerism status, the distributions of antigen-presenting cells (APCs), and T-cells in TLR4+/+ and TLR4-/- recipient mice. RESULTS: The general condition of TLR4-/- recipients was better than that of TLR4+/+ recipients, and the TLR4-/- recipient mice showed less severe GVHD manifestations than the TLR4+/+ recipient mice. Most of the APCs and T-cells in the host mouse spleen were derived from donor cells, and CD4+ T-cells, including memory T-cells, were in the majority in host mice. CONCLUSION: In general, our data show that TLR4 deletion attenuated GVHD development, which suggests that TLR4 could be used as a novel target and therapeutic paradigm in GVHD therapies.

ANTECEDENTES: La enfermedad de injerto contra huésped (EICH) es una complicación importante después del trasplante alogénico de células madre hematopoyéticas. OBJETIVOS: Para dilucidar el papel de TLR4, el principal receptor de LPS bacteriano, en el desarrollo de GVHD, construimos un modelo de GVHD en ratones knockout para TLR4 (TLR4-/-) y monitoreamos el quimerismo celular. MÉTODOS: En este estudio, usamos PCR para identificar si se establecieron ratones knockout para TLR4 (TLR4-/-). Antes del trasplante, pretratamos a los ratones con irradiación para obtener la dosis de irradiación adecuada. Se aplicó citometría de flujo para medir el estado de quimerismo, las distribuciones de APC y células T en ratones receptores TLR4+/+ y TLR4-/-. RESULTADOS: El estado general de los receptores de TLR4-/- fue mejor que el de los receptores de TLR4+/+, y los ratones receptores de TLR4-/- mostraron manifestaciones de GVHD menos graves que los ratones receptores de TLR4+/+. La mayoría de las APC y las células T en el bazo del ratón huésped se derivaron de las células del donante, y las células T CD4+, incluidas las células T de memoria, se encontraban en su mayoría en los ratones huéspedes. CONCLUSIÓN: En general, nuestros datos muestran que la eliminación de TLR4 atenuó el desarrollo de GVHD, lo que sugiere que TLR4 podría usarse como un nuevo objetivo y paradigma terapéutico en las terapias de GVHD.

Study of Cerebellar Tissue Following Induction of Aging in Mice By Di-Galactose / Estudio del Tejido Cerebeloso tras la Inducción del Envejecimiento en Ratones por D-Galactosa

SUMMARY: The cerebellum is a crucial area of the hindbrain that plays an essential role in balancing, excitement control, and subtle and accurate functions. Studies have shown that long-term use of D-galactose in mice, as with the symptoms of aging, causes morphological and functional disorders in the brain. This study was performed to evaluate the changes in the cerebellum cortex tissue and the measurement of reactive oxygen species (ROS) in the cerebellum following the induction of aging in mice by D-galactose. Accordingly, subjects were randomly assigned into two groups: Normal saline group and Aging group (D-galactose). To create an aging model, D- galactose, and saline solution (sodium chloride 0.9 %) were used. After completing the preparation and passage of the tissue, the cerebellum specimens were cut in 5 microns thickness and then stained with hematoxylin-eosin stain and finally examined under a Nikon microscope. Quantitative variables were analyzed by SPSS software using T-test. In the observations of cerebellum tissue samples, in the aged induced group by D-galactose, the most changes were observed in the Neuron purkinjense (Purkinje cells) layer. In the observations of the cerebellum tissue samples of aging group induced by D-galactose, the most changes were observed in the Neuron purkinjense, and the arrangement and placement of these cells were disorientated. The nucleus positioning was not central, and the Neuron purkinjense induced by aging were seen in different morphological forms. Necrosis, Chromatolysis, and Pyknosis were found. Based on the results, D-galactose (induction of aging) causes pathological changes in the cerebellar cortex, especially in the Neuron purkinjense layer.

El cerebelo es un área crucial del rombencéfalo que desempeña un papel esencial en el equilibrio, el control de la excitación y las funciones sutiles y precisas. Los estudios han demostrado que el uso a largo plazo de D-galactosa en ratones, al igual que con los síntomas del envejecimiento, provoca trastornos morfológicos y funcionales en el cerebro. Este estudio se realizó para evaluar los cambios en el tejido de la corteza del cerebelo y la medición de especies reactivas de oxígeno (ROS) en el cerebelo luego de la inducción del envejecimiento en ratones por D-galactosa. En consecuencia, los sujetos fueron asignados aleatoriamente a dos grupos: grupo de solución salina normal y grupo de envejecimiento (D-galactosa). Para crear un modelo de envejecimiento, se utilizaron D-galactosa y solución salina (cloruro de sodio al 0,9 %). Después de completar la preparación y el paso del tejido, las muestras de cerebelo se cortaron en un grosor de 5 µm y luego se tiñeron con tinción de hematoxilina-eosina y finalmente se examinaron bajo un microscopio Nikon. Las variables cuantitativas se analizaron mediante el software SPSS utilizando la prueba T. En las observaciones de muestras de tejido de cerebelo, en el grupo envejecido inducido por D-galactosa, la mayoría de los cambios se observaron en la capa de neuronas purkinjenses (células de Purkinje). En las observaciones de las muestras de tejido del cerebelo del grupo de envejecimiento inducidas por D-galactosa, la mayoría de los cambios se observaron en las neuronas purkinjenses, y la disposición y ubicación de estas células estaban desorientadas. El posicionamiento del núcleo no era central y las neuronas purkinjenses inducidas por el envejecimiento se observaban en diferentes formas morfológicas. Se encontró necrosis, cromatólisis y picnosis. Según los resultados, la D-galactosa (inducción del envejecimiento) provoca cambios patológicos en la corteza cerebelosa, especialmente en la capa de neuronas purkinjenses.

Efectos de la microvibración y estrógeno en la remodelación ósea: revisión sistemática / Effects of micro-vibration and estrogen on bone: a systematic review

Introducción: la pérdida de hueso es un suceso que afecta a la totalidad del esqueleto. Así, las alteraciones musculoesqueléticas afectan a millones de personas en todo el mundo y están entre las causas más comunes de dolor crónico. Objetivo: conocer los efectos de la microvibración y estrógeno en el remodelado óseo. Material y métodos: se realizó una revisión sistemática, se buscó en siete bases de datos, se incluyeron estudios clínicos controlados realizados con ratas o ratones en el periodo de publicación del 2004 al 2022. La calidad de la evidencia sintetizada se evaluó con la escala de Jadad. Resultados: se identificaron quince artículos como estudios primarios. La microvibración reportó cambios in vivo/in vitro totalmente dependientes del estímulo que conlleva incremento de la cortical externa. A su vez, con la administración de estrógeno se reportaron efectos, específicamente, en el hueso trabecular y en el periostio, así como colágeno inmaduro que indican un recambio óseo. Conclusión: tanto la microvibración como la administración de estrógeno coadyuvan a la remodelación del tejido óseo y son aprovechables como tratamiento en el momento que exista un problema de pérdida ósea (AU)

Introduction: Bone loss is an event that affects the entire skeleton. Thus, musculoskeletal disorders affect millions of people worldwide and are among the most common causes of chronic pain. Objective: to know the effects of micro-vibration and estrogen on bone remodelling. Material and methods: a systematic review was carried out; seven databases were searched; Controlled clinical studies conducted with rats or mice in the publication period from 2004 to 2022 were included. The quality of the synthesized evidence was assessed using the Jadad scale. Results: fifteen articles were identified as primary studies. Micro vibration reported in vivo/in vitro changes dependent on the stimulus that entails an increase in the outer cortex. In turn, with the administration of estrogen, effects were reported, specifically in the trabecular bone and in the periosteum, as well as immature collagen that indicates bone turnover. Conclusion: both micro-vibration and the administration of estrogen contribute to the remodelling of bone tissue and are usable as a treatment for bone loss (AU)

Age-related histopathological and biochemical testicular damages were ameliorated by vitamin C administration.

INTRODUCTION AND OBJECTIVES: Aging is an irreversible process associated with decreased biological functions that can lead to the reduction of reproductive organs capacities in males and females. Paternal age is a significant predictor of offspring health and development. So, the aim of this study was to evaluate the effects of vitamin C on histopathological and biochemical testicular changes following aging process with a focus on stereological methods. MATERIAL AND METHODS: For this study, 48 adult male NMRI mice were divided into two control and experimental groups. Mice in experimental group were supplemented with vitamin C (150mg/kg) including 24-h interval by oral gavage for 33 weeks. Same regime was performed for animals in control group except that vitamin C was replaced by water. Then, right testes were extracted for stereological and left testes were used for molecular analyses on weeks 8, 12, and 33. RESULTS: Our findings showed low semen quality, decreased level of serum Luteinizing hormone (LH), Follicle-stimulating hormone (FSH), and testosterone along with increased reactive oxygen species (ROS) production and higher apoptotic gene expression following aging. Stereological studies showed that the volume of testes, the length of seminiferous tubules, and the number of spermatogenic and none-spermatogenic cells decreased significantly during aging. Also, vitamin C consumption for 33 weeks significantly improved biochemical and histological indices. The impact of aging on male reproduction seems to be inevitable worldwide. Therefore, the use of protective and preventive remedies conserving male fecundity is very important and based on our results, vitamin C is a beneficial candidate for improving age-related testicular changes due to aging process.

[Improved immunity and mean lifespan in mature mice genetically deficient in catecholamine synthesis after living with wild type for two months]. / Mejoría de la inmunidad y la esperanza de vida en ratones maduros, genéticamente deficientes en la síntesis de catecolaminas, tras convivir con controles durante dos meses.

INTRODUCTION: Mice hemizygous in tyrosine hydroxylase (TH-HZ), the limiting enzyme in catecholamine synthesis, show premature immunosenescence, which in females is associated with a shorter lifespan than the corresponding controls (WT). The coexistence of TH-Hz with WT improves the immune function in both males and females in adulthood. OBJECTIVE: To test whether cohabitation for two months of mature male TH-HZ with WT improves the immune function of the former and whether this impacts the lifespan. MATERIAL AND METHODS: Mature male ICR-CD1 mice (13 ± 1 months) TH-HZ coexisted with WT (2:4 ratio in each cage) for two months. Peritoneal leukocytes were extracted from all animals at baseline, one month, and two months after cohabitation, and macrophage phagocytic capacity, macrophage and lymphocyte chemotaxis, natural killer (NK) antitumor activity, and lymphoproliferative capacity in response to the mitogens concanavalin A and lipopolysaccharide (LPS) were assessed. The animals were maintained under these conditions until their natural death. RESULTS: The TH-HZ, which start, in general, with lower values than the WT in the immune functions studied, improved them after two months of cohabitation, becoming similar to those of the controls. This improvement was already observed in NK activity after one month of cohabitation. The TH-HZ presented lower mean longevity than WT, but when they cohabited with WT, it was similar to the latter. CONCLUSION: The coexistence of TH-HZ male mice with WT mice for two months at mature age improves these genetically modified animals' immune response and longevity.

Anti-diabetic effects of Inonotus obliquus extract in high fat diet combined streptozotocin-induced type 2 diabetic mice.

Introduction: Introduction: type 2 diabetes (T2DM) is a complex disease affected by lifestyle and genetic factors. Although the drugs currently used to treat T2DM have certain curative effects, they still have some adverse side effects. Therefore, it is urgent to find new effective drugs with few side effects to cure T2DM. Objective: to study the role of Inonotus obliquus (IO) in diabetic model mice. Methods: we used high-fat diet (HFD) combined with streptozocin (STZ) to establish a diabetic mouse model. Mice were divided into non-high-fat diet group (ND), diabetes model group (HFD + STZ) and IO-treated diabetes model group (IO). The mice in the IO group were orally treated with IO (150 mg/kg) at 10 ml/kg for five weeks. Body weight, glucose level, food intake and water consumption, glucose tolerance and insulin tolerance were evaluated in all mice. The pathological sections of liver, kidney and pancreas were observed by hematoxylin-eosin staining. Results: after IO administration, the blood glucose level, water consumption, low-density lipoprotein (LDL) and triacylglycerol (TG) levels of mice decreased. Compared with the HFD + STZ group, the number of normal islet ß cells increased and focal necrosis of the liver was significantly reduced in the IO administration group. Conclusions: IO reduced the levels of blood glucose, restored body weight, and enhanced insulin sensitivity along with insulin tolerance and glucose tolerance in diabetic mice. Additionally, IO also reversed HFD and STZ-induced organ injury.

Introducción: Introducción: la diabetes mellitus tipo 2 (T2DM) es una enfermedad compleja influenciada por el estilo de vida y los factores genéticos. En la actualidad, aunque los medicamentos para la diabetes tipo 2 tienen cierto efecto curativo, todavía tienen algunos efectos secundarios. Por lo tanto, es urgente encontrar nuevos medicamentos para la diabetes tipo 2 que tengan un buen efecto curativo y menos efectos secundarios. Objetivo: estudiar el papel del Inonotus obliquus (IO) en ratones diabéticos. Métodos: se estableció un modelo de ratón diabético con dieta de alto contenido en grasas (HFD) y estreptozocina (STZ). Los ratones se dividieron en el grupo de dieta no alta en grasas (ND), el grupo modelo de diabetes mellitus (HFD + STZ) y el grupo modelo de diabetes mellitus tratado con IO. Los ratones del grupo IO recibieron 10 ml/kg de IO (150 mg/kg) durante cinco semanas. Se observaron el peso corporal, el nivel de azúcar en sangre, la ingesta de alimentos, la ingesta de agua potable, la tolerancia a la glucosa y la tolerancia a la insulina de los ratones de cada grupo, y se estudiaron muestras de biopsias hepáticas, renales y pancreáticas mediante tinción de hematoxilina eosina. Resultados: los niveles de glucosa en sangre, el consumo de agua, la lipoproteína de baja densidad (LDL) y los triglicéridos (TG) disminuyeron después de la administración de IO. En comparación con el grupo HFD+STZ, el número de células ß pancreáticas normales y la necrosis focal hepática disminuyeron significativamente en el grupo IO. Conclusiones: el IO redujo el nivel de glucosa en sangre, ayudó a recuperar el peso corporal y mejorar la sensibilidad a la insulina, la tolerancia a la insulina y la tolerancia a la glucosa en ratones diabéticos. Además, el IO revirtió el daño orgánico inducido por HFD y STZ.

Effect of voluntary wheel running on gut microbiota composition in developing mice.

Introduction: Background: exercise can increase the species and quantity of beneficial gut microbiota, enrich the diversity of microflora, and promote the development of symbiotic bacteria, especially in the stage of ontogeny. However, there is little evidence of the short-term voluntary exercise effect on the gut microbiota in developing mice. Material and method: therefore, we used short-term voluntary wheel running model to study the gut microbiota of developing mice (1 month old), and detected the fecal samples by 16S rRNA gene sequencing. Results: the results showed that after 4 weeks of voluntary wheel running, the body weight of the running group was significantly lower than that of the control group. Conclusion: there was a significant separation between the running group and the control group in beta diversity measures. At the family level, the clostridiales flora of the running group was higher than that of the control group. Compared with the control group, the abundance of parabacteroides flora and anaerovorax flora increased significantly, and the abundance of anaerotruncus flora and odoribacter flora decreased significantly in the running group. These results showed that gut microbiota be affected after short-term voluntary wheel running in developing mice.

Introducción: Introducción: el ejercicio puede aumentar las especies y la cantidad de microbiota intestinal beneficiosa, enriquecer la diversidad de la microflora y promover el desarrollo de bacterias simbióticas, especialmente en la etapa de ontogenia. Sin embargo, hay poca evidencia del efecto del ejercicio voluntario a corto plazo sobre la microbiota intestinal en ratones en desarrollo. Material y método: por lo tanto, utilizamos un modelo de carrera de ruedas voluntario a corto plazo para estudiar la microbiota intestinal de ratones en desarrollo (1 mes de edad) y detectamos las muestras fecales mediante la secuenciación del gen 16S rRNA. Resultados: los resultados mostraron que después de 4 semanas de carrera voluntaria con ruedas, el peso corporal del grupo de carrera fue significativamente más bajo que el del grupo de control. Conclusión: hubo una diferencia significativa entre el grupo de corredores y el grupo de control en las medidas de diversidad beta. A nivel familiar, la flora de clostridiales del grupo de corredores fue mayor que la del grupo de control. En comparación con el grupo de control, la abundancia de flora parabacteroides y flora anaerovorax aumentó significativamente, y la abundancia de flora anaerotruncus y flora odoribacter disminuyó significativamente en el grupo de carrera. Estos resultados mostraron que la microbiota intestinal se ve afectada después de la carrera voluntaria a corto plazo en ratones en desarrollo.

Membrana de cáscara de huevo para la curación de heridas superficiales en ratones / Eggshell membrane treatment for the healing of superficial open wounds in mice

Introducción. Las membranas de la cáscara de huevo presentan propiedades beneficiosas para la regeneración de tejidos y sus aplicaciones biomédicas son importantes. Objetivo. Demostrar la efectividad de las membranas de la cáscara de huevo no fecundado de gallina en el tratamiento de heridas abiertas superficiales en ratones, en comparación con el procedimiento convencional. Materiales y métodos. Se hizo una herida superficial lineal de 15 mm en la espalda de 10 ratones albinos machos. Los ratones se dividieron en cuatro grupos, uno no recibió ningún tratamiento y los otros tres sí: uno, tratamiento convencional, otro, con membranas de huevo directamente aplicadas a la herida y, el otro, con membranas en forma de polvo. La evolución de las heridas se registró en fotografías y se calculó la tasa de reducción de la longitud de la herida, así como el tiempo y el porcentaje de curación. Los porcentajes de curación se analizaron con ANOVA y la prueba de Dunnett (p<0,05). Resultados. Con los tratamientos con membranas de huevo y polvo de membrana, se logró una tasa de reducción de longitud de 1.009 y 1.020 mm/día, respectivamente, y un tiempo de curación de 12 días, en tanto que, con el tratamiento convencional, la tasa de reducción fue de 0,852 mm/día y la curación se dio en 16 días. El análisis estadístico mostró diferencias significativas entre los tratamientos con membrana de huevo y el tratamiento convencional. Conclusiones. Las membranas de la cáscara de huevo aplicadas de forma directa y en polvo resultaron más efectivas que la aplicación del procedimiento convencional en el tratamiento de heridas abiertas superficiales en ratones.

Introduction: Eggshell membranes offer beneficial properties for tissue regeneration and their biomedical applications are important. Objective: To demonstrate the effectiveness of hen unfertilized eggshell membranes in the treatment of superficial open wounds in mice compared to the conventional procedure. Materials and methods: 15-mm linear wounds were made on the back of 10 male CF-1 mice. The mice were divided into four groups. One group did not receive any treatment and the other three were given conventional treatment, eggshell membranes dressing, and eggshell membranes powder. The evolution of the wounds was registered by photographs. We measured wounds length and healing time and calculated the length reduction rate and the percentage of healing. The healing percentages were analyzed by ANOVA with a post hoc Dunnett test (p<0.05). Results: The treatments with eggshell membranes and membrane powder had a wound-length reduction rate of 1.009 and 1.020 mm/day, greater than the conventional treatment, of 0.852 mm/day, and a healing time of 12 days, i.e., less than the 16-day conventional treatment. The statistical analysis showed significant differences between eggshell membrane treatments and the conventional treatment. Conclusions: Eggshell membrane dressing and eggshell membrane powder applied to open superficial wounds in mice were more effective than the conventional treatment in regenerating injured tissue in mice.

miR-125a-5p in astrocytes attenuates peripheral neuropathy in type 2 diabetic mice through targeting TRAF6.

INTRODUCTION: Elimination or blocking of astrocytes could ameliorate neuropathic pain in animal models. MiR-125a-5p, expressed in astrocyte derived extracellular vesicles, could mediate astrocyte function to regulate neuron communication. However, the role of miR-125a-5p in DPN (diabetic peripheral neuropathy) remains elusive. MATERIALS AND METHODS: Type 2 diabetic mouse (db/db) was used as DPN model, which was confirmed by detection of body weight, blood glucose, mechanical allodynia, thermal hyperalgesia, glial fibrillary acidic protein (GFAP) and monocyte chemoattractant protein-1 (MCP-1). Astrocyte was isolated from db/db mouse and then subjected to high glucose treatment. The expression of miR-125a-5p in db/db mice and high glucose-induced astrocytes was examined by qRT-PCR analysis. Downstream target of miR-125a-5p was clarified by luciferase reporter assay. Tail vein injection of miR-125a-5p mimic into db/db mice was then performed to investigate role of miR-125a-5p on DPN. RESULTS: Type 2 diabetic mice showed higher body weight and blood glucose than normal db/m mice. Thermal hyperalgesia and mechanical allodynia were decreased in db/db mouse compared with db/m mouse, while GFAP and MCP-1 were increased in db/db mouse. High glucose treatment enhanced the protein expression of GFAP and MCP-1 in astrocytes. Sciatic nerve tissues in db/db mice and high glucose-induced astrocytes exhibited a decrease in miR-125a-5p. Systemic administration of miR-125a-5p mimic increased mechanical allodynia and thermal hyperalgesia, whereas it decreased GFAP and MCP-1. TRAF6 (tumor necrosis factor receptor associated factor 6) was validated as target of miR-125a-5p. CONCLUSION: MiR-125a-5p in astrocytes attenuated DPN in db/db mice by up-regulation of TRAF6, which indicated the potential therapeutic effect.

Mice as an animal model for the study of adipose tissue and obesity / Ratones como modelo animal para el estudio del tejido adiposo y la obesidad

SUMMARY: The study of adipose tissue has gained increasing importance in the biomedical area due to its implications for health and obesity. Obesity is a situation of great concern mainly in the Western world due to its high prevalence and morbidity. Experimental studies on obesity often need a model where it is possible to carry out experiments, drug testing, and other therapeutic procedures, which are typically not possible in humans. Although several animals are used for obesity studies, rodents are by far the most used animals, and among rodents, mice are particularly indicated for this investigation. This mini review will introduce the challenging classification of obesity in rodents, paralleling human obesity, defining and classifying what an obese mouse is. The text will differentiate between white adipose tissue (WAT, aimed at endocrine secretion and lipogenesis) and brown adipose tissue (BAT, aimed at thermogenesis) and describe the browning process of white adipocytes in an adaptation to increase thermogenesis. The text will also describe the various types of body fat in mice with their differences and indications for investigation and teach how to recognize and dissect these fats. At the end of this introductory reading, the young researcher is expected to have acquired sufficient knowledge to start an experimental investigative project on obesity.

RESUMEN: El estudio del tejido adiposo ha ganado una importancia creciente en el área biomédica por sus implicaciones para la salud y la obesidad. La obesidad es una situación de gran preocupación, principalmente, en el mundo occidental debido a su alta prevalencia y morbilidad. Los estudios experimentales sobre la obesidad a menudo necesitan un modelo en el que sea posible realizar experimentos, pruebas de drogas y otros procedimientos terapéuticos, que normalmente no son posibles en humanos. Aunque se utilizan varios animales para estudios de obesidad, los roedores son, con mucho, los animales más utilizados y, entre los roedores, los ratones están especialmente indicados para esta investigación. Esta mini revisión presenta la desafiante clasificación de la obesidad en roedores, en paralelo con la obesidad humana, definiendo y clasificando qué es un ratón obeso. El texto diferencia entre tejido adiposo blanco (WAT, destinado a la secreción endocrina y lipogénesis) y tejido adiposo marrón (BAT, destinado a la termogénesis) y describe el proceso de pardeamiento de los adipocitos blancos en una adaptación para aumentar la termogénesis. El texto también describe los diversos tipos de grasa corporal en ratones con sus diferencias e indicaciones para la investigación y enseña cómo reconocer y diseccionar estas grasas. Al final de esta lectura introductoria, se espera que el joven investigador haya adquirido los conocimientos suficientes para iniciar un proyecto de investigación experimental sobre la obesidad.

Up-regulation of Arl4a gene expression by broccoli aqueous extract is associated with improved spermatogenesis in mouse testes / La regulación positiva de la expresión del gen Arl4a por acción del extracto acuoso de brócoli se asocia con una mejor espermatogénesis en testículos de ratón

Abstract | Introduction: Broccoli (Brassica oleracea) is well known for its properties as an anticancer, antioxidant, and scavenger of free radicals. However, its benefits in enhancing spermatogenesis have not been well established. Objective: To study broccoli aqueous extract effects on sperm factors and the expression of genes Catsper1, Catsper2, Arl4a, Sox5, and Sox9 in sperm factors in mice. Materials and methods: Male mice were divided randomly into six groups: (1) Control; (2) cadmium (3 mg/kg of mouse body weight); (3) orally treated with 200 µl broccoli aqueous extract (1 g ml-1); (4) orally treated with 400 µl of broccoli aqueous extract; (5) orally treated with 200 broccoli aqueous extract plus cadmium, and (6) orally treated with 400 µl of broccoli aqueous extract plus cadmium. We analyzed the sperms factors and Catsper1, Catsper2, Arl4a, Sox5, and Sox9 gene expression. Results: An obvious improvement in sperm count and a slight enhancement in sperm motility were observed in mice treated with broccoli extract alone or with cadmium. Sperm viability was reduced by broccoli extract except for the 200 µl dose with cadmium, which significantly increased it. Interestingly, Arl4a gene expression increased in the 400 µl broccoli- treated group. Likewise, the Arl4a mRNA level in mice treated with cadmium and 200 µl of broccoli extract was higher than in the cadmium-treated mice. Furthermore, broccoli extract enhanced the mRNA level of Catsper2 and Sox5 genes in mice treated with 200 µl and 400 µl broccoli extract plus cadmium compared with the group treated solely with cadmium. Conclusion: The higher sperm count in broccoli-treated mice opens the way for the development of pharmaceutical products for infertile men.

Resumen | Introducción. El brócoli (Brassica oleracea) se conoce por sus propiedades como anticancerígeno, antioxidante y eliminador de radicales libres. Sin embargo, sus beneficios en la espermatogénesis aún no se han determinado suficientemente. Objetivo. Estudiar los efectos del extracto acuoso de brócoli sobre los factores espermáticos y la expresión de los genes Catsper1, Catsper2, Arl4a, Sox5 y Sox9 en ratones. Materiales y métodos. Los ratones machos se dividieron aleatoriamente en seis grupos: 1) control; 2) tratados con cadmio, 3 mg/kg de peso corporal; 3) tratados con 200 µl de extracto acuoso de brócoli (1 g ml-1); 4) tratados con 400 µl de extracto acuoso de brócoli; 5) tratados con 200 µl de extracto acuoso de brócoli más cadmio, y 6) tratados con 400 µl de extracto acuoso de brócoli más cadmio. El extracto acuoso de brócoli se administró por vía oral. Se analizaron los factores espermáticos y la expresión de los genes Catsper1, Catsper2, Arl4a, Sox5 y Sox9. Resultados. Se observó una mejoría obvia en el recuento y una ligera mejoría en la motilidad de los espermatozoides, en ratones tratados con extracto de brócoli solo o con cadmio. La viabilidad de los espermatozoides se redujo con el extracto de brócoli, excepto con la dosis de 200 µl más cadmio, la cual la aumentó significativamente. Curiosamente, la expresión del gen Arl4a aumentó en el grupo tratado con 400 µl del extracto. Asimismo, el ARNm del Arl4a en ratones tratados con cadmio y 200 µl del extracto, fue más abundante que en los ratones tratados solo con cadmio. Además, el extracto de brócoli aumentó la cantidad de ARNm de los genes Catsper2 y Sox5 en ratones tratados con 200 y 400 µl de extracto de brócoli más cadmio, en comparación con el grupo tratado únicamente con cadmio. Conclusión. El mayor número de espermatozoides en ratones tratados con brócoli abre el camino al desarrollo de productos farmacéuticos para hombres infértiles.

Tartrazina induce genotoxicidad en linfocitos de BALB/c Mus musculus / Tartrazine induces genotoxicity in BALB/c Mus musculus lymphocytes

RESUMEN Objetivos. Determinar el efecto genotóxico de la tartrazina en linfocitos de sangre periférica de Mus musculus BALB/c. Materiales y métodos. Se realizó un estudio experimental, a través de cinco grupos, con cinco ratones en cada uno. Se les registró el peso durante 17 semanas y, en la semana 15 se les administró suero fisiológico (control negativo), dicromato de potasio 25 mg/kg de peso corporal (pc) (control positivo) y tartrazina a dosis de 0,75 mg/kg pc, 7,5 mg/kg pc y 75 mg/kg pc, durante siete días, a excepción del control positivo que fue en dosis única. Luego, cada 24 h se obtuvo una muestra de sangre periférica de la cola y se realizó el frotis, secado y coloración. Posteriormente, se realizó el conteo de 1000 linfocitos por muestra de cada ratón, en todos los tratamientos. Resultados. Los tres tratamientos con tartrazina no causaron diferencias significativas en el peso de ratones a la semana 15, pero sí produjeron diferencias significativas en la frecuencia de linfocitos micronucleados, siendo el tratamiento con tartrazina de 75 mg/kg pc el de mayor efecto genotóxico, induciendo un promedio de 1,63 ± 0,08 linfocitos micronucleados, comparado con el control positivo que generó un promedio de 1,42 ± 0,08 linfocitos micronucleados. Conclusiones. La tartrazina produjo un efecto genotóxico, incrementando el número de linfocitos micronucleados, a dosis de 0,75; 7,5 y 75 mg/kg pc y no afecta el peso corporal durante siete días de administración en M. musculus BALB/c.

ABSTRACT Objectives. To determine the genotoxic effect of tartrazine on peripheral blood lymphocytes of BALB/c Mus musculus. Materials and methods. An experimental study was carried out using five groups, with five mice in each group. Their weight was registered for 17 weeks, and at week 15 they were administered physiological saline solution (negative control), potassium dichromate at 25 mg/kg body weight (bw) (positive control) and tartrazine at doses of 0.75 mg/kg bw, 7.5 mg/kg bw and 75 mg/kg bw, for seven days, with the exception of the positive control which was a single dose. Then, every 24 hours, a peripheral blood sample was obtained from the tail, which was then smeared, dried and stained. Subsequently, 1000 lymphocytes were counted for each sample from each mouse, for all treatment groups. Results. The three tartrazine treatments did not cause significant differences in the weight of mice at week 15, but did produce significant differences in the frequency of micronucleated lymphocytes, with the 75 mg/kg bw tartrazine treatment having the greatest genotoxic effect, inducing an average of 1.63 ± 0.08 micronucleated lymphocytes, compared to the positive control which obtained an average of 1.42 ± 0.08 micronucleated lymphocytes. Conclusions. Tartrazine produced a genotoxic effect, increasing the number of micronucleated lymphocytes, at doses of 0.75; 7.5 and 75 mg/kg bw and did not affect body weight during seven days of administration to BALB/c M. musculus.

Determination of the temperature causing a nociceptive response in the tail of albino BALB/c mice.

INTRODUCTION: Designs for determining nociceptive response in rodents are of great use in neurology and experimental neuroscience. Immersing mice's tails in warm water is one of the most widely used procedures to evaluate this response; however, a wide range of temperatures are used in different studies. Knowing the temperature that produces a powerful nociceptive response in the tail of BALB/c mice is extremely useful. METHODS: Eight 2-month-old male BALB/c mice were used. A 14-cm high beaker was filled with water up to 13cm. The animals' tails were immersed in the container with a starting temperature of 36°C. The water temperature was raised in 1°C increments until we identified the temperatures that produced nociceptive responses. That response was determined by counting the time taken before the mouse shook its tail to remove it from the water. RESULTS: Six of the 8 mice began shaking their tails at the temperature of 51°C. All animals removed their tails from the water at the temperatures of 54°C, 55°C, and 56°C, taking a mean time of 8.54, 7.99, and 5.33seconds, respectively. ANOVA applied to the response times for each of the 3 temperatures indicated revealed a value of F=2.8 (P=.123). CONCLUSIONS: The response time was statistically similar for the temperatures of 54°C, 55°C, and 56°C; however, the data were less dispersed for the latter temperature.

Spatiotemporally differential expression of HER (human epithelial growth factor receptor)-2 with sexual dimorphism in submandibular gland of mice

SUMMARY: Considering that the submandibular gland (SMG) of postnatal mice performs active cell proliferation, apoptosis and differentiation which are regulated by proto-oncogene products in cancerous cells, the expression and localization of a proto-oncogene product HER (human epidermal growth factor receptor)-2 was examined in SMG of postnatal mice. In Western blot analysis, the expression for HER-2 was high until pre-puberty, and it decreased from puberty to young adult stages with male SMG more dominant. In immunohistochemistry, the immunoreactivity was positive in acinar and ductal cells of newborn SMG with distinct localization at the intercellular apposition sites. The immunoreactivity in acinar cells progressively decreased to negligible levels by pre-pubertal stage, while it remained positive in most ductal cells throughout the postnatal time-course. The immunoreactivity in cells of terminal tubules and intercalated ducts, both of which have a high potential to produce cells, were seen at levels similar to those of more proximal ducts, while the immunoreactivity in ductal basal cells was significantly high, but the granular convoluted tubule cells were seen at negligible levels in male and at faint levels in female. In immuno-electron microscopy of excretory ducts, the immunoreactivity was dominantly localized on the basal infolding membranes as well as vesicles and vacuoles of various sizes, but rarely in Golgi apparatus and mitochondria. The immunoreactivity without association to any membranous structures were also seen, though not numerous. The relation of expression levels of HER-2 in various portions of normal SMG to those in their cancerous ones is briefly discussed.

RESUMEN: Considerando que la glándula submandibular (GSM) de ratones postnatales realiza la proliferación celular activa, apoptosis y diferenciación que están reguladas por productos protooncogénicos en células cancerosas, la expresión y localización de un producto protooncogénico HER (receptor del factor de crecimiento epidérmico humano) - 2 se examinó en GSM de estos ratones. En el análisis de Western blot, la expresión de HER-2 fue alta hasta la prepubertad, y disminuyó desde la pubertad hasta las etapas de adultos jóvenes con GSM macho más dominante. En inmunohistoquímica, la inmunorreactividad fue positiva en las células acinares y ductales de GSM de recién nacido con una localización distinta en los sitios de aposición intercelular. La inmunorreactividad en las células acinares disminuyó progresivamente a niveles insignificantes en la etapa prepuberal, mientras que permaneció positiva en la mayoría de las células ductales durante el transcurso del tiempo posnatal. La inmunorreactividad en las células de los túbulos terminales y los conductos intercalados, los cuales tienen un alto potencial para producir células, se obser- vó a niveles similares a los de los conductos más proximales, mientras que la inmunorreactividad en las células basales ductales fue significativamente alta, pero en el túbulo contorneado granular las células se observaron en niveles insignificantes en los machos y en niveles débiles en las hembras. En la microscopía inmunoelectrónica de los conductos excretores, la inmunorreactividad se localizó de manera predominante en las membranas de pliegues basales, así como en vesículas y vacuolas de varios tamaños, pero raramente en el aparato de Golgi y en las mitocondrias. También se observó la inmunorreactividad sin asociación a ninguna estructura membranosa, aunque no numerosa. Se discute brevemente la relación de los niveles de expresión de HER-2 en varias porciones de GSM normal con aquellos en sus cancerosos.

Anticonvulsant effects of Paeonia daurica subsp. macrophylla root extracts in pentylenetetrazol-induced seizure models in mice.

INTRODUCTION: In the present study, anticonvulsant effects of aqueous extract (AE), hydro-alcoholic crude extract (HE), and its fractions (F-CHCl3, F-EtOAc, F-MeOH) of Paeonia daurica subsp. macrophylla (P. daurica ssp. macrophylla) root examined by using a pentylenetetrazol-induced model (PTZ) on mice. METHODS: HE and its fractions as well as AE, in concentrations of (100, 200 and 400mg/kg), valproate (Val) (100 and 200mg/kg), and saline (negative control) (10mg/kg) were injected intraperitoneally (i.p.) 30min before PTZ (80mg/kg, i.p.). The time taken before the onset of myoclonic convulsions (MC), MC duration, time taken before the onset of generalized tonic-clonic seizures (GTCS), the duration of GTCS, and the percentage of GTCS and mortality protection recorded. The plant's anticonvulsant mechanisms were assessed using flumazenil (5mg/kg, i.p.) before AE (100, 200, and 400mg/kg, i.p.) injection. GraphPad Prism software was used to compare the differences between various treatment groups with one-way analysis of variance (ANOVA) followed by Tukey-Krammer multiple comparison tests. RESULTS: All the plant samples except F-EtOAc significantly delayed the onset and decreased the duration of PTZ-induced MCS and GTCS, and significantly reduced the GTCS and mortality rate. Pretreatment with flumazenil diminished the significant anticonvulsant effects of AE against PTZ-induced seizures. CONCLUSIONS: It can report that extract of P. daurica ssp. macrophylla might be a helpful guide for future studies in the treatment of epilepsy.

Leishmania (L.) amazonensis infection impairs reproductive and fetal parameters in female mice / La infección por Leishmania (L.) amazonensis afecta parámetros reproductivos y fetales en ratones hembras

ABSTRACT Leishmaniasis is a group of parasitic zoonotic diseases caused by intracellular protozoans belonging to the genusLeishmania. Little is known about the effects that this parasitosis may have on the reproductive parameters and pregnancy of infected humans and pets. This study aimed to evaluate the influence of chronic cutaneous leishmaniasis caused byLeishmania (Leishmania) amazonensison reproductive and fetal parameters using a female murine model. A control group of female BALB/c mice and a group infected withL. (L.) amazonensiswere mated with healthy males. Clinical parameters were monitored during the pre-mating and gestational periods. Female mice were euthanized on day 19 of gestation, when the fetuses were weighed and their length measured and embryonic resorptions and fetal death were recorded. We observed five fetal deaths and three embryonic resorptions in the infected group. Furthermore, there was a decrease in fertility in the infected group (26.32%). The weight of the offspring from infected mothers was lower than that in the control group (1.019±0.035g and 1.163±0.032g,p<0.01). Fetal length was reduced in the infected group (3.71±0.05cm in the control group and 3.40±0.06cm in the infected groupp<0.001). This study shows that cutaneous leishmaniasis caused byL. (L.) amazonensisimpairs reproductive and fetal parameters in mice.

RESUMEN La leishmaniasis comprende un grupo de enfermedades zoonóticas parasitarias causadas por protozoos intracelulares pertenecientes al géneroLeishmania. Poco se conoce sobre los efectos que esta parasitosis puede tener sobre los parámetros reproductivos y la gestación en humanos y en otras especies infectadas. El objetivo de este estudio fue determinar la influencia de la leishmaniasis cutánea crónica, causada porLeishmania (Leishmania) amazonensis, en parámetros reproductivos y fetales. Se apareó un grupo control y un grupo infectado de ratones hembra BALB/c (previamente inoculado conL. (L.) amazonensis) con machos sanos. Se analizaron parámetros clínicos durante los períodos pregestacional y gestacional. Las hembras fueron eutanasiadas en el día 19 de gestación, momento en el cual se pesaron y midieron los fetos y se registraron las reabsorciones embrionarias y las muertes fetales. Se observaron 5 muertes fetales y 3 reabsorciones embrionarias en el grupo infectado. Además, hubo una disminución en la fertilidad de este último grupo (26,32%). Por otra parte, el peso de la descendencia de madres infectadas fue menor que el del grupo control (1,019±0,035g y 1,163±0,032g, respectivamente,p<0,01). Por último, la longitud fetal se redujo en el grupo infectado (3,71±0,05cm en el grupo control y 3,40±0,06cm en el grupo infectado,p<0,001). Este estudio muestra que la leishmaniasis cutánea causada porL. (L.) amazonensisafecta los parámetros reproductivos y fetales en ratones.

Evaluación de la toxicidad de chalconas sintéticas con potencial anti-Leishmania en ratones BALB/c / Toxicity assessment of synthetic chalcones with antileishmanial potential in BALB/c mice

RESUMEN Objetivo: Evaluar la toxicidad de tres chalconas sintéticas administradas por vía intraperitoneal en ratones BALB/c. Materiales y métodos: La dosis letal media (DL50) se estimó por el método Up-and-Down de Dixon. La toxicidad subcrónica de las chalconas se evaluó a 20 y 40 mg/kg por 21 días. Se evaluó el efecto tóxico a nivel de comportamiento, fisiológico, bioquímico e histológico. Resultados: La chalcona 43 generó moco en las heces, daño visceral (hígado) y alteración en el coeficiente de órganos (riñón, p = 0,037 y cerebro, p = 0,008) en comparación con el grupo control. Además, en el análisis histológico se observó que esta chalcona produjo edema, inflamación y necrosis en los órganos evaluados, aunque no hubo diferencia significativa con el control. Todos los parámetros bioquímicos no difirieron significativamente entre los grupos de tratamiento a dosis de 40 mg/kg y el control. Conclusiones: La DL50 para las tres chalconas fue superior a 550 mg/kg de peso corporal. Las chalconas 40 y 42 son relativamente no tóxicas. Ambas pueden considerarse seguras para la aplicación vía intraperitoneal en ratones BALB/c y, en consecuencia, son posibles candidatas para ser usadas en el tratamiento contra las leishmaniosis.

ABSTRACT Objective: To evaluate the toxicity of three synthetic chalcones administered intraperitoneally to BALB/c mice. Materials and methods: The median lethal dose (LD50) was estimated by Dixon's Up-and-Down method. Subchronic toxicity of chalcones was evaluated at 20 and 40 mg/kg for 21 days. Behavioral, physiological, biochemical, and histological toxic effects were evaluated. Results: Chalcone 43 produced mucus in feces, visceral damage (liver) and alterations in organ coefficient (kidney, p = 0.037 and brain, p = 0.008) when compared to the control group. In addition, histological analysis showed that this chalcone produced edema, inflammation and necrosis in the evaluated organs, although there was no significant difference with the control. None of the biochemical parameters differed significantly between the treatment groups at 40 mg/kg dose and the control. Conclusions: The LD50 for all three chalcones was greater than 550 mg/kg of body weight. Chalcones 40 and 42 were found to be relatively non-toxic. Both can be considered safe for intraperitoneal application in BALB/c mice and, consequently, are potential candidates for use in the treatment of leishmaniasis.

miR-125a-5p in astrocytes attenuates peripheral neuropathy in type 2 diabetic mice through targeting TRAF6.

INTRODUCTION: Elimination or blocking of astrocytes could ameliorate neuropathic pain in animal models. MiR-125a-5p, expressed in astrocyte derived extracellular vesicles, could mediate astrocyte function to regulate neuron communication. However, the role of miR-125a-5p in DPN (diabetic peripheral neuropathy) remains elusive. MATERIALS AND METHODS: Type 2 diabetic mouse (db/db) was used as DPN model, which was confirmed by detection of body weight, blood glucose, mechanical allodynia, thermal hyperalgesia, glial fibrillary acidic protein (GFAP) and monocyte chemoattractant protein-1 (MCP-1). Astrocyte was isolated from db/db mouse and then subjected to high glucose treatment. The expression of miR-125a-5p in db/db mice and high glucose-induced astrocytes was examined by qRT-PCR analysis. Downstream target of miR-125a-5p was clarified by luciferase reporter assay. Tail vein injection of miR-125a-5p mimic into db/db mice was then performed to investigate role of miR-125a-5p on DPN. RESULTS: Type 2 diabetic mice showed higher body weight and blood glucose than normal db/m mice. Thermal hyperalgesia and mechanical allodynia were decreased in db/db mouse compared with db/m mouse, while GFAP and MCP-1 were increased in db/db mouse. High glucose treatment enhanced the protein expression of GFAP and MCP-1 in astrocytes. Sciatic nerve tissues in db/db mice and high glucose-induced astrocytes exhibited a decrease in miR-125a-5p. Systemic administration of miR-125a-5p mimic increased mechanical allodynia and thermal hyperalgesia, whereas it decreased GFAP and MCP-1. TRAF6 (tumor necrosis factor receptor associated factor 6) was validated as target of miR-125a-5p. CONCLUSION: MiR-125a-5p in astrocytes attenuated DPN in db/db mice by up-regulation of TRAF6, which indicated the potential therapeutic effect.