Histology and histopathology Vol.32,nº12 (2017)
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- PublicationOpen AccessBariatric surgery influences β-Cell turnover in non obese rats(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Camacho Ramírez, Alonso; Blandino Rosano, Manuel; Segundo Iglesias, M. Carmen; Lechuga Sancho, Alfonso M.; Aguilar Diosdado, Manuel; Pérez Arana, Gonzalo M.; Prada Oliveira, J. ArturoBackground. The aim of this study was to investigate the relation between the different bariatric surgeries and pancreatic β-cell turnover. Material and Methods. We used healthy adult male Wistar rats to undergo the different techniques. Three surgical techniques were developed (malabsorptive, Sleeve gastrectomy and Roux-Y Gastric Bypass-), together with two control groups (Sham and fasting control). Pancreatic β-cell mass was measured, as well as apoptosis, proliferation and neogenesis related to cellular turnover. Otherwise, we measured the functional issues to elucidate the physiological role that these surgical techniques trigger in the carbohydrate metabolism (e.g. food intake, weight gain, intraperitoneal glucose tolerance test, and basal glycaemia). Results included the differences in phenotypes of the rat after the surgery. The rats did not show important differences in glycaemic parameters between the surgical groups. The β-cell mass presented modifications related with proliferation processes. A significant increase of β-cell mass in the malabsorptive technique was reported. On the other hand, the peripheral resistance to insulin tended to be reduced in rats which underwent malabsorptive and mixed techniques. Conclusion. This work showed an increase in β-cell mass after the resection of an important portion of small bowel. The Roux-Y Gastric Bypass produced a non-significant increase in β-cell mass. We considered that these implications of surgery over the endocrine pancreas must be one of the mechanisms related to the improvement of type 2 Diabetes mellitus following bariatric surgery.
- PublicationOpen AccessSulfur dioxide: foe or friend for life?(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Wang, Xin Bao; Cui, Hong; Liu, Xiao Hong; Du, Jun BaoSulfur dioxide (SO2) is a toxic gas and air pollutant. The toxic effects of SO2 have been extensively studied. Oxidative damage due to SO2 can occur in multiple organs. Inhaled SO2 can also cause chromosomal aberrations, DNA damage and gene mutations in mammals. However, SO2 can also be generated from the sulfur-containing amino acid, Lcysteine. Recent studies have shown that SO2 has a vasorelaxant effect, and ameliorates pulmonary hypertension and vascular remodeling. SO2 can also reduce lung injury and myocardial injury in rats. In addition, SO2 reduces myocardial ischemia-reperfusion injury and atherosclerotic lesions. Therefore, SO2 exerts both detrimental and protective effects in mammals. Is SO2 a foe or friend for life?
- PublicationOpen AccessResveratrol decreases FoXO protein expression through PI3K-Akt-dependent pathway inhibition in H2O2-treated synoviocytes(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Xu, Bin; Wang, Gaoyuan; Zhang, Junqiang; Cao, Wei; Chen, XiaoyuThe aim of this study was to investigate the effects of resveratrol (Res) on hydrogen peroxide (H2O2)- treated fibroblast-like synoviocytes (FLSs) in vitro. We studied the phosphoinositide 3-kinase (PI3K)-Akt pathway inhibition-mediated effects of Res on forkhead box O (FoXO) mRNA expression levels. FLS viability was determined by Cell Counting Kit-8 (CCK-8) assay, and FLS apoptosis was measured by terminal deoxyribo nucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining and flow cytometry. FoXO1, FoXO3 and FoXO4 mRNA expression levels in FLSs were determined by RT-PCR, and p-Akt, Akt, p-FoXO1, FoXO1, p-FoXO3, FoXO3, Bcl-2 and Bax protein expression levels were determined by western blotting. Our results showed that low H2O2 concentrations (20 μM) can promote FLS growth and that Res significantly inhibited FLS activity. Moreover, Res significantly increased the number of apoptotic cells and the ratio of Bax/Bcl-2 protein expression in the group treated with Res compared with the group treated with H2O2 and LY294002, a PI3K inhibitor. Res also decreased FoXO1, FoXO3 and FoXO4 mRNA expression levels and pAkt/Akt, p-FoXO1/FoXO1, p-FoXO3/FoXO3 protein expression levels. Taken together, these findings indicate that Res can induce apoptosis in H2O2-treated FLSs in part by inhibiting the PI3K-Akt signaling pathway.
- PublicationOpen AccessExpression profiles of angiogenesis in two high grade chondrosarcomas: A xenotransplant experience in nude mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Giner, Francisco; López Guerrero, José Antonio; Machado, Isidro; García Casado, Zaida; Fernández Serra, Antonio; Peydró Olaya, Amando; Llombart Bosch, AntonioBackground. Chondrosarcomas (Chs) are malignant cartilage-forming tumors that represent the third most common malignant solid tumor of bone in adults. Angiogenesis is a major factor for tumor growth and metastasis. Our aim was to make a histological, immunohistochemical, ultrastructural and molecular characterization of the neovascularization established between xenotransplanted Chs and the host during the initial phases of growth in nude transfer, in order to find potential markers for distinguishing between high grades II and III Chs. Methods. two xenotransplanted high grade human Chs were evaluated. Tumor pieces were implanted subcutaneously on the backs of 14 athymic Balb-c nude mice. The animals were sacrificed 24, 48, and 96 hours; and 7, 14, 21 and 28 days after implantation. Two grade I Chs were also transferred in nude but did not grow. Results. Morphological differences were apparent between these two Chs during the early stages of neoplastic growth. Immunohistochemistry demonstrated overexpression of pro-angiogenic factors 24h-48h after tumor implantation. Additionally, neoplastic cells co-expressed chemokines (CXCL9, CXCL10 and GRO) and their receptors. Molecular studies showed two expression profiles, revealing an early and a late phase in the angiogenic process. Conclusion. High grade Chs demonstrated two different stages of induced angiogenesis, with an intimate association between structural and molecular events that might explain the different aggressive biological behavior of grade II and III Chs. The present model may be useful for testing the effect of anti-angiogenic drugs.
- PublicationOpen AccessGestational protein restriction: Study of the probable effects on cardiac muscle structure and function in adult rats(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Amer, Mona G.; Mohamed, Nader M.; Shaalan, Aly A.M.Intrauterine growth restriction (IUGR) has been linked to heart disease in adulthood. This study aimed to examine the effect of gestational protein restriction during fetal and early postnatal life on the cardiac muscle structure and function in adult offspring. Pregnant female rats were randomly divided into two dietary groups: normal-protein diet (NP) and low-protein diet (LP). Fifteen male offspring from each group were included in the study. Offspring body weights were recorded at birth and monthly from weaning until 24 weeks of age while systolic blood pressure was measured weekly. At the end of the experiment, hearts were weighed and processed for light and electron microscopy and immunohistochemical study. Immunohistochemical staining for localization of inducible nitric oxide synthase (iNOS) and connexin 43 proteins was performed. The gestational protein restriction induced deleterious effects on adult offspring including decreased birth weight, heart weight, and heart rate, and increased systolic blood pressure. Histologically, the number of cardiomyocytes decreased and cardiac fibrosis increased. Signs of degeneration at both structural and ultra-structural levels of cardiomyocytes were also seen. The iNOS was up regulated in LP offspring which was a promoter for apoptosis, while connexin 43 was down regulated which would affect heart conductivity and contractility. Our results demonstrate that adult offspring body weight and cardiac muscle structure and function can be programmed by maternal gestational nutrition. These adverse outcomes suggest the criticality of dietary behavior during pregnancy on long-term offspring cardiac health.
- PublicationOpen AccessHuman sperm motility, capacitation and acrosome reaction are impaired by 2-arachidonoylglycerol endocannabinoid(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Francou, M.M.; Girela, Jose Luis; de Juan, A.; Ten, J.; Bernabeu Mora, Roberto; De Juan, J.The endocannabinoids are cannabinoids synthesized by mammalian tissues. These compounds are closely related to the regulation of the male reproductive system. However, little is known about the effects produced by 2-arachidonoylglycerol (2AG) on in vitro human sperm functions. This study was undertaken to determine the effects produced by 2AG on fresh human sperm and in the capacitation technique. Semen samples from healthy young men were exposed to different concentrations of 2AG before and during capacitation technique. In this work, we have demonstrated that 2AG induces the spontaneous acrosome reaction and reduces progressive motility in fresh human sperm. During the capacitation technique, sperm becomes more sensitive to low concentrations of 2AG, triggering the acrosome reaction and inhibiting protein phosphorylation. In summary, 2AG affects the in vitro functionality of human sperm by reducing motility, inhibiting capacitation and triggering the acrosome reaction.
- PublicationOpen AccessThe dexamethasone induced osteogenic differentiation of dental follicle cells(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Morsczeck, Christian; Reichert, Torsten E.Mesenchymal stem cells are excellent for in vitro studies about biological processes during the differentiation of osteogenic progenitor cells into mineralizing cells such as osteoblasts. Human dental follicle cells (DFCs) are dental mesenchymal stem cells and they can be isolated from third molar teeth. Because DFCs are the genuine progenitor cells of periodontal tissue cells, they have been used for the evaluation of molecular mechanisms during the differentiation of undifferentiated stem cells into alveolar osteoblasts and cementoblasts. To reveal molecular mechanisms of osteogenic differentiation, initial studies investigated the proteome and the transcriptome of DFCs after the induction of the osteogenic differentiation with the glucocorticoid dexamethasone. These studies showed for example that dexamethasone induces the transcription factor ZBTB16 (zinc finger and BTB domain containing protein 16) and that ZBTB16 is crucial for osteogenic differentiation of DFCs. This article is a survey of the molecular mechanisms in DFCs during osteogenic differentiation with dexamethasone.
- PublicationOpen AccessMorphofunctional basis of the different types of angiogenesis and formation of postnatal angiogenesis-related secondary structures(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Díaz Flores, L.; Gutierrez, R.; García Suárez, M.P.; Sáez, F.J.; Gutiérrez, E.; Valladares, F.; Carrascosa, J.L.; Díaz Flores Jr, L.; Madrid Cuevas, Juan FranciscoWe review the morpho-functional basis of the different types of angiogenesis and report our observations, including the formation of angiogenesisrelated secondary structures. First of all, we consider the following issues: a) conceptual differences between angiogenesis and vasculogenesis, b) incidence of angiogenesis in pre- and postnatal life, c) regions of vascular tree with angiogenic capacity, d) cells (endothelial cells, pericytes, CD34+ adventitial stromal cells of the microvasculature and inflammatory cells) and extracellular matrix components involved in angiogenesis, e) events associated with angiogenesis, f) different types of angiogenesis, including sprouting and intussusceptive angiogenesis, and other angiogenic or vascularization forms arising from endothelial precursor cells (postnatal vasculogenesis), vasculogenesis mimicry, vessel co-option and piecemeal angiogenesis. Subsequently, we consider the specific morphofunctional characteristics of each type of angiogenesis. In sprouting angiogenesis, we grouped the events in three phases: a) activation phase, which includes vasodilation and increased permeability, EC, pericyte and CD34+ adventitial stromal cell activation, and recruitment and activation of inflammatory cells, b)sprouting phase, encompassing EC migration (concept and characteristics of endothelial tip cells, tip cell selection, lateral inhibition, localized filopodia formation, basal lamina degradation and extracellular changes facilitating EC migration), EC proliferation (concept of endothelial stalk cells), pericyte mobilization, proliferation, recruitment and changes in CD34+ adventitial stromal cells and inflammatory cells, tubulogenesis, formation of a new basal lamina, and vascular anastomosis with capillary loop formation, and c) vascular remodelling and stabilization phase (concept of phalanx cells). Subsequently, the concept, incidence, events and mechanisms are considered in the other forms of angiogenesis. Finally, we contribute the formation of postnatal angiogenesis-related secondary structures: a) intravascular structures through piecemeal angiogenesis, including intravascular papillae in vessel tumours and pseudotumours (intravascular papillary endothelial hyperplasia, vascular transformation of the sinus in lymph nodes, papillary intralymphatic angioendothelioma or Dabska tumour, retiform hemangioendothelioma, hemangiosarcoma and lymphangiosarcoma), vascular septa in hemorrhoidal veins and intravascular projections in some tumours; b) arterial intimal thickening; c) intravascular tumours and pseudotumours (e.g. intravenous pyogenic granulomas and intravascular myopericytoma); d) vascular glomeruloid proliferations; and e) pseudopalisading necrosis in glioblastoma multiform.
- PublicationOpen AccessIncreased annexin A2 and decreased β-catenin in adenomyosis contribute to adenomyosis-associated dysmenorrhea(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Liu, Li Xue; Wu, Yin Ga; Zheng, JianObjective. To investigate the expression of annexin A2 (ANXA2) and β-catenin in eutopic and ectopic endometrium, and their relationships with adenomyosis-associated dysmenorrhea. Methods. From December 2013 to June 2014, ectopic endometrium (n=30) and eutopic endometrium (n=30) of adenomyosis were collected as experimental group, and endometrium (n=30) of uterine myoma as control group from the department of gynecology and obstetrics, the affiliated hospital of Inner Mongolia medical university. The expression of ANXA2 and βcatenin was detected by immunohistochemical S-P method, followed by the Pearson correlations for the correlation analysis of ANXA2 and β-catenin with adenomyosis-associated dysmenorrhea. Meanwhile, the levels of preoperative serum ANXA2 of patients with adenomyosis (n=42) and uterine myoma (n=42) were also measured by enzyme-linked immunosorbent assay (ELISA). Results. Immunohistochemistry and ELISA identified a higher expression of ANXA2 in eutopic and ectopic endometrium of adenomysis tissues, whereas βcatenin protein was down-regulated. Furthermore, there was a significant positive correlation between ANXA2 expression and dysmenorrhea degree, while there was a negative linear correlation between β-catenin expression and dysmenorrhea degree in ectopic endometrium. Conclusion. These results suggested that increased ANXA2 and less expressed β-catenin were correlated to adenomyosis-associated dysmenorrhea. It may provide a new idea of diagnosis and treatment to adenomyosisassociated dysmenorrhea.
- PublicationOpen AccessTreatment with a selective histone deacetylase 6 inhibitor decreases lupus nephritis in NZB/W mice(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Vieson, Miranda D.; Gojmerac, Alexander M.; Khan, Deena; Dai, Rujuan; van Duzer, John H.; Mazitschek, Ralph; Caudell, David L.; Liao, Xiaofeng; Luo, Xin M.; Reilly, Christopher M.To date, there are 18 histone deacetylase (HDAC) enzymes, divided into four classes, which alter protein function by removing acetyl groups from lysine residues. Prior studies report that non-selective HDAC inhibitors decrease disease in lupus mouse models. Concern for adverse side effects of non-selective HDAC inhibition supports investigation of selective-HDAC inhibition. We hypothesized that a selective HDAC-6 inhibitor (HDAC6i) will alleviate disease in a mouse model of lupus by increasing acetylation of alphatubulin. Intraperitoneal injections of the selective HDAC6i ACY-1083 (0.3 mg/kg, 1 mg/kg, or 3 mg/kg), vehicle control, or dexamethasone were administered to 21-week-old, female NZB/W mice, 5 days a week, for 13 weeks. Disease progression was evaluated by proteinuria, serum levels of anti-dsDNA antibody, cytokines and immunoglobulins, and post mortem evaluation of nephritis and T cell populations in the spleen. HDAC6i treatment decreased proteinuria, glomerular histopathology, IgG, and C3 scores when compared to vehicle-treated mice. Within glomeruli of HDAC6i-treated mice, there was increased acetylation of alpha-tubulin and decreased NF-κB. Additionally, HDAC6i decreased serum IL-12/IL-23 and Th17 cells in the spleen. Taken together, these results suggest HDAC6 inhibition may decrease lupus nephritis in NZB/W mice via mechanisms involving acetylation of alphatubulin and decreased NF-κB in glomeruli as well as inhibition of Th17 cells.