RESUMO
SUMMARY: The R-spondin protein family is a group of proteins that enhance Wnt/b-catenin signaling and have pleiotropic functions in stem cell growth and development. In the literature reviews, there is no histomorphological study showing the localization and distribution of R-spondins in different hypothalamic nuclei. For this reason, the purpose of this study was to determine the localization, distribution characteristics, and densities in the hypothalamic nuclei of neurons expressing Rspo1 and Rspo3 proteins. The free-floating brain sections of the male rats who were not exposed to any treatment were stained with the indirect immunoperoxidase method using the relevant antibodies. As a result of the immunohistochemical studies, it was determined that neurons expressing the Rspo1 protein were found in large numbers in the supraoptic nucleus (SON), the suprachiasmatic nucleus (SCh), anterior paraventricular nucleus, periventricular hypothalamic nucleus (PeV), anterior hypothalamic area, magnocellular preoptic nucleus (MCPO) and the lateral hypothalamic area (LH) from the hypothalamic nuclei, while they were localized in fewer numbers in the arcuate nucleus (ARC). Rspo3 protein expression was found in neurons localized in the hypothalamic nuclei SON, paraventricular nucleus (PVN), PeV, ARC, ventromedial nucleus (VMH), LH, anterior parvicellular nucleus, and zona inserta (ZI). In addition, neurons synthesizing both peptides were found in the cortex and hippocampus regions (H). Rspo1 and 3 proteins are expressed in hypothalamic energy homeostatic areas, thus these proteins may be involved in the regulation of food intake.
La familia de proteínas R-espondina es un grupo de proteínas que mejoran la señalización de Wnt/b-catenina y tienen funciones pleiotrópicas en el crecimiento y desarrollo de las células madre. En las revisiones de la literatura no existen estudios histomorfológicos que muestren la localización y distribución de las R-espondinas en diferentes núcleos hipotalámicos. Por esta razón, el propósito de este estudio fue determinar la localización, características de distribución y densidades en los núcleos hipotalámicos de neuronas que expresan las proteínas Rspo1 y Rspo3. Secciones de cerebro flotantes de ratas macho que no fueron expuestas a ningún tratamiento se tiñeron con el método de inmunoperoxidasa indirecta utilizando los anticuerpos pertinentes. Como resultado de los estudios inmunohistoquímicos, se determinó que las neuronas que expresan la proteína Rspo1 se encontraron en gran número en el núcleo supraóptico (SON), el núcleo supraquiasmático (SCh), el núcleo paraventricular anterior, el núcleo hipotalámico periventricular (PeV), el núcleo hipotalámico anterior área, núcleo preóptico magnocelular (MCPO) y el área hipotalámica lateral (LH) de los núcleos hipotalámicos, mientras que se localizaron en menor número en el núcleo arqueado (ARC). La expresión de la proteína Rspo3 se encontró en neuronas localizadas en los núcleos hipotalámicos SON, núcleo paraventricular (PVN), PeV, ARC, núcleo ventromedial (VMH), LH, núcleo parvicelular anterior y zona inserta (ZI). Además, se encontraron neuronas que sintetizan ambos péptidos en las regiones de la corteza y el hipocampo (H). Las proteínas Rspo1 y 3 se expresan en áreas homeostáticas de energía hipotalámicas, por lo que estas proteínas pueden estar involucradas en la regulación de la ingesta de alimentos.
Assuntos
Animais , Masculino , Ratos , Trombospondinas/metabolismo , Hipotálamo/metabolismo , Imuno-Histoquímica , Ratos Sprague-DawleyRESUMO
Objective To investigate the role and mechanism of RSPO1 in osteoblasts differentiation.Methods The xCELLigence system was used to study the toxicity and role of RSPO1 on the C2C12 cells proliferation.Alkaline phosphatase (ALP) activity was measured by using a phosphatase detection kit.The expression of osteoprotegerin (OPG) was determined using enzyme-linked immunosorbent assay (ELISA).Wnt/β-catenin signaling was evaluated using the TOPflash T cell factor (TCF) luciferase reporter assay.Western blotting assay was performed to confirm the accumulation of β-catenin protein.T test was used for statistical analysis.Results RSPO1 had no effect on the C2C12 cells proliferation,and it produced no toxicity to C2C12 cells.RSPO1 alone resulted in a slight increase in the activity of ALP (2.85±0.08 vs 1.74±0.21,t=3.014,P＜0.05) and the expression of OPG (1.29±0.13 vs 1.00±0.21,t=3.348,P＜0.05),whereas the combination of RSPO1 and Wnt3A significantly amplified ALP activity (81.37±5.08 vs 1.74±0.21,t=31.31,P＜0.01) and OPG protein expression (5.26±0.60 vs 1.00±0.21,t=6.99,P＜0.01).RSPO1 synergized with Wnt3A to activate TCF activity and induce accumulation of β-catenin (3.28±0.18 vs 1.00±0.21,t=10.94,P＜0.05).However,RSPO1 alone had no effect on the TCF activity and β-catenin accumulation (1.25±0.08 vs 1.00±0.21,t=2.225,P＞0.05).Conclusion Our study has revealed that RSPO1 synergized with Wnt3A in activating Wnt/β-catenin signaling pathway to induce osteoblasts differentiation,which demonstrate the therapeutic potential of RSPO1 for RA.
RESUMO
Objective To explore the expression and effects of RSp1 and ?-catenin in the intestinal epithelium with intestinal ischemia-reperfusion injury(IIRI)in of mouse.Methods Fifty healthy male kunming mice were randomly divided into control group(n=10)and experimental group(n=40).All mice in control group were only subjected to laparotomy,while the other mice underwent 20 minutes of intestinal mesenteric artery occlusion followed by 6 hours(group A),12 hours(group B),24 hours(group C)and 48 hours(group D)of reperfusion.RT-PCR was used to detect RSpo1 and ?-catenin in small intestine in intestinal ischemia-reperfusion groups and in control group.Results The villous heights of intestinal in experimental groups were significantly lower than that in control group(P