Therapeutic Effect of HDAC5 Binding and Cell Penetrating Peptide for the Treatment of Inflammatory Bowel Disease

BACKGROUND@#Inflammatory bowel disease (IBD) is an incurable disease that negatively influences the quality of life of patients. Current and emerging therapies target proinflammatory cytokines and/or receptors to downregulate proinflammatory responses, but insufficient remission requires other therapeutic agents. Herein, we report that the synthetic antiinflammatory peptide 15 (SAP15) is capable of cell penetration and anti-inflammatory activity in human macrophages. @*METHODS@#SAP15 was labeled with fluorescence and administered to human leukemia monocytic cells (THP-1) cells for cell penetration analysis. Using biolayer interferometry analysis, the binding affinity of SAP15 with histone deacetylase 5 (HDAC5) was measured. SAP15-treated THP-1 cells were analyzed by protein phosphorylation assay, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). In addition, in vivo analysis of the therapeutic effect on IBD was observed in a dextran sulfate sodium (DSS)-induced model. Samples from SAP15-treated mice were analyzed at both the macroscopic and microscopic levels using ELISA, myeloperoxidase (MPO) assays, and histological evaluations. @*RESULTS@#SAP15 was internalized within the cytosol and nucleus of THP-1 cells and bound to the HDAC5 protein. SAP15-treated macrophages were assessed for protein phosphorylation and showed inhibited phosphorylation of HDAC5 and other immune-related proteins, which led to increased M2-like macrophage markers and decreased M1-like macrophage markers and tumor necrosis factor-a and interleukin-6 cytokine levels. The SAP15 treatment on IBD model showed significant recovery of colon length. Further histological analysis of colon demonstrated the therapeutic effect of SAP15 on mucosal layer. Moreover, proinflammatory cytokine levels and MPO activity from the plasma show that SAP15 is effective in reduced proinflammatory responses. @*CONCLUSION@#These findings suggest that SAP15 is a novel peptide with a novel cell-penetrating peptide with antiinflammatory property that can be used as a therapeutic agent for IBD and other inflammatory diseases.

Nitrosylation of b2-Tubulin Promotes Microtubule Disassembly and Differentiated Cardiomyocyte Beating in Ischemic Mice

BACKGROUND@#Beating cardiomyocyte regeneration therapies have revealed as alternative therapeutics for heart transplantation. Nonetheless, the importance of nitric oxide (NO) in cardiomyocyte regeneration has been widely suggested, little has been reported concerning endogenous NO during cardiomyocyte differentiation. @*METHODS@#Here, we used P19CL6 cells and a Myocardiac infarction (MI) model to confirm NO-induced protein modification and its role in cardiac beating. Two tyrosine (Tyr) residues of b2-tubulin (Y106 and Y340) underwent nitrosylation (Tyr-NO) by endogenously generated NO during cardiomyocyte differentiation from pre-cardiomyocyte-like P19CL6 cells. @*RESULTS@#Tyr-NO-b2-tubulin mediated the interaction with Stathmin, which promotes microtubule disassembly, and was prominently observed in spontaneously beating cell clusters and mouse embryonic heart (E11.5d). In myocardial infarction mice, Tyr-NO-b2-tubulin in transplanted cells was closely related with cardiac troponin-T expression with their functional recovery, reduced infarct size and thickened left ventricular wall. @*CONCLUSION@#This is the first discovery of a new target molecule of NO, b2-tubulin, that can promote normal cardiac beating and cardiomyocyte regeneration. Taken together, we suggest therapeutic potential of Tyr-NO-b2-tubulin, for ischemic cardiomyocyte, which can reduce unexpected side effect of stem cell transplantation, arrhythmogenesis.

Optimization of Microenvironments Inducing Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Endothelial Cell-Like Cells

BACKGROUND: Stem cell engineering is appealing consideration for regenerating damaged endothelial cells (ECs) because stem cells can differentiate into EC-like cells. In this study, we demonstrate that tonsil-derived mesenchymal stem cells (TMSCs) can differentiate into EC-like cells under optimal physiochemical microenvironments.METHODS: TMSCs were preconditioned with Dulbecco's Modified Eagle Medium (DMEM) or EC growth medium (EGM) for 4 days and then replating them on Matrigel to observe the formation of a capillary-like network under light microscope. Microarray, quantitative real time polymerase chain reaction, Western blotting and immunofluorescence analyses were used to evaluate the expression of gene and protein of EC-related markers.RESULTS: Preconditioning TMSCs in EGM for 4 days and then replating them on Matrigel induced the formation of a capillary-like network in 3 h, but TMSCs preconditioned with DMEM did not form such a network. Genome analyses confirmed that EGM preconditioning significantly affected the expression of genes related to angiogenesis, blood vessel morphogenesis and development, and vascular development. Western blot analyses revealed that EGM preconditioning with gelatin coating induced the expression of endothelial nitric oxide synthase (eNOS), a mature EC-specific marker, as well as phosphorylated Akt at serine 473, a signaling molecule related to eNOS activation. Gelatin-coating during EGM preconditioning further enhanced the stability of the capillary-like network, and also resulted in the network more closely resembled to those observed in human umbilical vein endothelial cells.CONCLUSION: This study suggests that under specific conditions, i.e., EGM preconditioning with gelatin coating for 4 days followed by Matrigel, TMSCs could be a source of generating endothelial cells for treating vascular dysfunction.

Intracellular Remodeling and Accumulation of Aberrant Lysosomes in Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Parathyroid-Like Cells

Differentiation of mesenchymal stem cells (MSC) into a variety of cell lineages such as adipocytes, osteocytes, and chondrocytes is often accompanied up-regulation of autophagy. In our study, we demonstrated that the expression of autophagy-associated proteins (p-Beclin 1, LC3A, LC3B, p-AMPK, p-mTOR and ATG3, ATG7, and ATG12-5) over a period of time was hardly distinguishable from control tonsil-derived MSC (TMSC). Despite the unnoticeable difference in autophagy activation between differentiated TMSC (dTMSC) and the control (cTMSC), we reported significant changes in intracellular compositions in differentiated TMSC into functional parathyroid-like cells secreting parathyroid hormone (PTH). By using transmission electron microscopy (TEM), we observed accumulation of multivesicular bodies (MVB) comprising small, degraded compartments densely accumulated as dark granular or amorphous clumps, multilamellar bodies and lipid droplets in dTMSC. However, no such structures were found in cTMSC. These results suggest that differentiation of TMSC into parathyroid-like cells producing PTH hormone is hardly dependent on autophagy activation in the beginning of our conditions. Furthermore, our results of intracellular remodeling and accumulated endo-lysosomal storage bodies in the later stages of TMSC differentiation present a possible role of the structures in PTH secretion.

The Correlation of Usual Dietary Isoflavone Intake and Serum Osteocalcin / 대한지역사회영양학회지

Few studies have conducted the bone health benefits of usual dietary isoflavone intake in Korean college-student women. To elucidate this benefitial effects and correlations between dietary isoflavone and nutrients intake and bone formation marker (ALPase, osteocalcin), questionnaires, anthropometric measurements, serum mineral (Ca, P, Mg) concentrations were analyzed. Fifty three subjects were used in this study. The average age, height, body weight, BMI, body fat content and triceps skinfold thickness were 21.43 year, 161.07 cm, 52.81 kg, 20.48 kg/m2, 20.72% and 17.59 mm respectively. Soy food intake frequencies were soybean > soybean curd > soypaste stew > soybean milk. The average calorie, protein and Ca intake were 1766.21 +/- 62.54 kcal, 66.45 +/- 2.00 g and 549.62 +/- 27.55 mg respectively. Serum ALPase activity and osteocalcin concentration were 115.74 +/- 3.6 u/L and 7.15 ng/ml respectively. Usual dietary isoflavone intake was positively related to calory, protein, Ca, P intake and serum Ca, Mg concentration. Serum osteocalcin concentration was negatively correlated with isoflavone intake (r2=0.28, p < 0.05). In these results, usual dietary isoflavone can support an additive effect to bone health and Ca nutrition.

Concentrations of Major Minerals and Trace Elements in Sera of The Breast-fed and Formula-fed Infants / 대한지역사회영양학회지

The serum concentrations of the major minerals(calcium, phosphorus, magnesium, sodium, potassium) and trace elements(iron, Zinc, copper manganese)were datermined in 23 breast fed infants living in Eumsung, Choong-buk region. The results obtained are summarized as follows: 1)The mean levels of calcium, phosphorus, magnesium, sodium and potassium of the serum of total subjects were 8.15+/-0.33mg/dI, 11.06+/-0.16mg/dI, 2.00+/-0.14mg/dI, 3.4476+/-17.99mg/dI and 9.06+/-2.04mg/dI respectively. 2)The serum concentrations of iron, zinc, copper and manganese in total subjects averaged 95.83+/-0.33mg/dI, 93.79+/-7.06 microgram/dI and 98.57+/-7.06 microgram/dI and 4.93+/-0.62 microgram/dI respecitively. 3)Breast fed infants had significantly higher serum calcim, magnesium, sodium and iron concentrations than the formula fed groups. Otherwise, serum potassium, copper and manganese levels in breast milk fed infants were significantly lower than those in formula fed infants. 4) In formula fed infants, serum potassium and copper levels increased increased significantly with months after birth.

Ecological Studies of Maternal-Infant Nutrition and Feeding in Urban Low Income Areas - II. Anthropometric Measurements, Dietary Intakes, Breast Milk Components and Serum Lipid/Fatty Acid Composition of Lactating Women / 대한지역사회영양학회지

This study was aimed at evaluating the nutritional status of Korean lactating women, who were attending peripheral community clinics in low-income areas, by anthropometric measurement, estimating dietary intakes and analyzing the contents of breast milk components, serum lipid content and fatty acid composition. Regarding anghropometric measurements, maternal circumferences decreased significantly at the trunk but not at the limbs. The skinfold thickness on all regions had the tendency to decrease during lactation. It was found that intakes of energy, protein, lipid and carbohydrates were desirable while calcium, zinc, copper and vitamin A were low and sodium was excessive. The energy % of fat was 22.41% of the total energy which is higher than the RDA for adults but it is proper for lactating women, but energy % from each fatty acid was usually low and an intake ratio of omega6/omega3 fatty acids was desirable. Accordingly, it is thought that dietary fat intake was balanced in quality. Concentrations of the nutrients in matured human milk showed the same range as that reported form different regions with the exception of calcium and magnesium which were relatively low. Serum lipid contents were decreased with the time postpartum and the average serum lipid content of lactating women were similar to the normal range of adults. Serum polyunsaturated fatty acids. especially omega6 fatty acids have gradually increased with the time postpartum, but the composition were not greatly influenced by dietary intakes.