The outcomes of dexmedetomidine and calcitriol on flap viability

Abstract Purpose: To evaluate protective effects of dexmedetomidine, calcitriol and their combination. Methods: Forty Wistar-albino rats were divided into 4 groups; group of Sham (Group Sham); group of dexmedetomidine (Group DEX); group of calcitriol (Group CAL) and group of dexmedetomidineandcalcitriol (Group DEX-CAL). Photographic analysis was used for macroscopic analysis and perfusion analyses were evaluated by scintigraphy. Additionally, tissue malondialdehyde (MDA) and total oxidant status (TOS) and total antioxidant activity (TAS) were recorded and oxidative stress index (OSI) was calculated. Each flap was assessed by histopathology. Results: Compared to Group Sham, the viable flap areas were higher in all treatment groups both by photographic image analyses and perfusion analyses (p<0.05). Group DEX-CAL had the highest viable flap percentage both in scintigraphic and photographic analyses; whereas Group Sham had the lowest viable flap percentage. Similarly, TAS and MDA levels were elevated and TOS levels were declined in all treatment groups compared to Group Sham (p<0.005). Histopathological analysis at flap demarcation zone confirmed neovascularization was significantly higher and edema, necrosis and inflammation were significantly lower in all treatment groups compared to Group Sham. Conclusion: The outcomes show that additional premedication with either dexmedetomidine or calcitriol or their combination reduces ischemia-reperfusion injury of flap area and show significant increase in the percentage of viable flap tissue.

Effects of 1, 25-dihydroxyvitamin D3 on experimental periodontitis and AhR/NF-κB/NLRP3 inflammasome pathway in a mouse model

Abstract Vitamin D has been known to have important regulatory functions in inflammation and immune response and shows inhibitory effects on experimental periodontitis in animal models. However, the potential mechanism has yet to be clarified. Recent studies have highlighted Aryl hydrocarbon receptor (AhR) and its downstream signaling as a crucial regulator of immune homeostasis and inflammatory regulation. Objective: This study aimed to clarify the effect of 1,25-dihydroxyvitamin D3 (VD3) on experimental periodontitis and AhR/nuclear factor-κB (NF-κB)/NLR pyrin domain-containing 3 (NLRP3) inflammasome pathway in the gingival epithelium in a murine model. Methodology: We induced periodontitis in male C57BL/6 wild-type mice by oral inoculation of Porphyromonas gingivalis (P. gingivalis), and subsequently gave intraperitoneal VD3 injection to the mice every other day for 8 weeks. Afterwards, we examined the alveolar bone using scanning electron microscopy (SEM) and detected the gingival epithelial protein using western blot analysis and immunohistochemical staining. Results: SEM images demonstrated that alveolar bone loss was reduced in the periodontitis mouse model after VD3 supplementation. Western blot analyses and immunohistochemical staining of the gingival epithelium showed that the expression of vitamin D receptor, AhR and its downstream cytochrome P450 1A1 were enhanced upon VD3 application. Additionally, VD3 decreased NF-κB p65 phosphorylation, and NLRP3, apoptosis-associated speck-like protein, caspase-1, interleukin-1β (IL-1β) and IL-6 protein expression. Conclusions: These results implicate the alleviation of periodontitis and the alteration of AhR/NF-κB/NLRP3 inflammasome pathway by VD3 in the mouse model. The attenuation of this periodontal disease may correlate with the regulation of AhR/NF-κB/NLRP3 inflammasome pathway by VD3.

Impact of 1 25(OH) 2 D 3 on TG content in liver of rats with type 2 diabetes

Abstract Purpose: To evaluate the effects of 1,25 dihydroxy vitamin D3 (1,25(OH)2D3) on the content of triglyceride (TG), as well as on the gene and protein expressions of adiponectin receptor 2 (AdipoR2), p38 mitogen-activated protein kinase (P38MAPK), and lipoprotein lipase (LPL) in the liver of rats with type 2 diabetes mellitus (T2DM) so as to provide theoretical basis for exploring the mechanism by which 1,25(OH)2D3 regulates TG. Methods: Wistar rats were divided into four groups (n=25), with different treatments and detected the gene and protein expressions of AdipoR2, p38MAPK, and LPL in the liver tissue by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Meanwhile, the content of TG in the liver tissue was detected by the Enzyme-linked immunosorbent assay. Results: The expression of AdipoR2, p38MAPK, LPL gene and protein in the liver of VitD intervention group was significantly higher than that in T2DM group (P <0.05), while the TG content was significantly lower than that in T2DM group (P <0.05). Conclusion: 1,25(OH)2D3 can decrease the content of TG in the liver, and its mechanism may be achieved by upregulating the expressions of AdipoR2, p38MAPK, and LPL in the liver.

Anti-Proliferative and Apoptotic Activities of Mullerian Inhibiting Substance Combined with Calcitriol in Ovarian Cancer Cell Lines

PURPOSE: This study aimed to investigate whether Mullerian inhibiting substance (MIS) in combination with calcitriol modulates proliferation and apoptosis of human ovarian cancer (OCa) cell lines (SKOV3, OVCAR3, and OVCA433) and identify the signaling pathway by which MIS mediates apoptosis. MATERIALS AND METHODS: OCa cell lines were treated with MIS in the absence or presence of calcitriol. Cell viability and proliferation were evaluated using the Cell Counting Kit-8 assay and apoptosis was evaluated by DNA fragmentation assay. Western blot and enzyme-linked immunosorbent assay were used to determine the signaling pathway. RESULTS: The cells showed specific staining for the MIS type II receptor. Treatment of OCa cells with MIS and calcitriol led to dose- and time-dependent inhibition of cell growth and survival. The combination treatment significantly suppressed cell growth, down-regulated the expression of B-cell lymphoma 2 (Bcl-2), and up-regulated the expressions of Bcl-2 associated X protein, caspase-3, and caspase-9 through the extracellular signal-regulated kinase signaling pathway. CONCLUSION: These results, coupled with a much-needed decrease in the toxic side effects of currently employed therapeutic agents, provide a strong rationale for testing the therapeutic potential of MIS, alone or in combination with calcitriol, in the treatment of OCa.

Treatment with 1,25(OH)2D3 induced HDAC2 expression and reduced NF-κB p65 expression in a rat model of OVA-induced asthma

Recent evidence indicates that a deficiency of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) may influence asthma pathogenesis; however, its roles in regulating specific molecular transcription mechanisms remain unclear. We aimed to investigate the effect of 1,25(OH)2D3 on the expression and enzyme activity of histone deacetylase 2 (HDAC2) and its synergistic effects with dexamethasone (Dx) in the inhibition of inflammatory cytokine secretion in a rat asthma model. Healthy Wistar rats were randomly divided into 6 groups: control, asthma, 1,25(OH)2D3 pretreatment, 1,25(OH)2D3 treatment, Dx treatment, and Dx and 1,25(OH)2D3 treatment. Pulmonary inflammation was induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Inflammatory cells and cytokines in the bronchoalveolar lavage (BAL) fluid and histological changes in lung tissue were examined. Nuclear factor kappa B (NF-κB) p65 and HDAC2 expression levels were assessed with Western blot analyses and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Enzyme activity measurements and immunohistochemical detection of HDAC2 were also performed. Our data demonstrated that 1,25(OH)2D3 reduced the airway inflammatory response and the level of inflammatory cytokines in BAL. Although NF-κB p65 expression was attenuated in the pretreatment and treatment groups, the expression and enzyme activity of HDAC2 were increased. In addition, 1,25(OH)2D3 and Dx had synergistic effects on the suppression of total cell infusion, cytokine release, and NF-κB p65 expression, and they also increased HDAC2 expression and activity in OVA/OVA rats. Collectively, our results indicated that 1,25(OH)2D3 might be useful as a novel HDAC2 activator in the treatment of asthma.

Regulation of matrix metalloproteinase-9 protein expression by 1alpha,25-(OH)2D3 during osteoclast differentiation

To investigate 1alpha,25-(OH)2D3 regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1alpha,25-(OH)2D3 during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1alpha,25-(OH)2D3 inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1alpha,25-(OH)2D3 administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.

[Induction of specific T-cell response following in vivo treatment of dendritic cells by 1,25-dihydroxycholecalciferol]

Dendritic cells [DCs], as the managers of the immune response, have a crucial role in forming the direction and nature of the immune response. Some compounds such as 1,25-dihydroxycholecalciferol affect the function of DCs and can be used to shift the immune functions toward favorite directions. The aim of this study was to investigate the in vivo effects of 1, 25- dihydroxycholecalciferol on DCs surface markers, their potential to induce specific T-cell responses and the cytokines profile. 1, 25-dihidroxycholecolciferol was regularly injected intraperitoneal into C57BL/6 mice. DCs were separated from the spleens of calciferol treated and non-treated mice using magnetic beads. The expression of DCs surface markers was investigated by flow cytometric analysis. The separated cells were pulsed by myelin oligodendrocyte glycoprotein [MOG] and injected subcutaneously into front footpads of syngeneic mice. After five days, the lymphocytes from regional lymph nodes were separated and used for the lymphocyte transformation test [LTT] and determination of the interferon gamma/interleukin 4 [IFN gamma/IL-4] ratio by ELISA technique. Statistical analysis of the obtained results showed reduced expression of maturation markers and co-stimulatory molecules by cholecalciferol treated DCs. The specific T-cell stimulation potential of treated DCs as well as the induced IFN gamma/IL-4 ratio was also down-regulated compared to non-treated cells [p value<0.05]. It seems that 1,25-dihydroxycholecolciferol can regulate the DCs function and maturation state in vivo. The T-cell stimulation rate and Th1/Th2 cytokines ratio also changes following interaction with cholecalciferol treated DCs

Calcium metabolism in cows receiving an intramuscular injection of 1,25-dihydroxyvitamin D3 combined with prostaglandin F2alpha closely before parturition

To determine the effect of exogenous 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] combined with induced parturition on calcium (Ca) metabolism, cows received a single intramuscular injection of 1,25(OH)2D3 and prostaglandin F2alpha (PGF2alpha) closely before calving. Ten late-pregnant, multiparous Holstein cows were assigned to 1,25(OH)2D3 group (five treated with both 1,25(OH)2D3 and PGF2alpha) and control group (five treated with PGF2alpha). 1,25(OH)2D3 group showed an increase in plasma Ca concentration around parturition, whereas control group revealed a decrease in plasma Ca level. Plasma Ca concentration in 1,25(OH)2D3 group were significantly higher than that in control group during .0.5 to 3 days after parturition.

Potential regulation of PTH/PTHrP receptor expression in choriocarcinoma cells

Parathyroid hormone-related peptide [PTHrP] have been found to be expressed in a variety of human tumors. Parathyroid hormone-related peptide is known as the major mediator of humoral hypercalcemia of malignancy, may also regulate placental calcium flux, uterine contraction and fetal tissue development. The purpose of this study is to evaluate the expression of PTH/PTHrP receptor in choriocarcinoma JAR cell line. This study was carried out at the Department of Biochemistry, College of Science, King Saud University, Riyadh, Kingdom of Saudi Arabia, between November 2002 and August 2003. Choriocarcinoma JAR cell line treated for 12 and 72 hours with epidermal growth factor, [EGF] [20ng/ml], estradiol, E2 [10-8 M], dexamethasone, [DEX] [10-8 M] or 1,25 dihydroxycholecalciferol, 1,25 [DHCC] [10-8 M]. We investigated the expression of parathyroid hormone [PTH]/PTHrP receptor in JAR cell line with these treatments compared with untreated JAR cells. The PTH/PTHrP receptor expression were detected with 3.3nM 125I-PTHrP-34Tyrosine. The expression of the receptors at 12 hours were increased following exposure to EGF, E2 or DEX, whereas 1,25 DHCC inhibited the receptor expression. In further experiments at 72 hours with the same treatments, the receptors expression were remarkably increased with EGF, E2 or DEX, whereas, 1,25 DHCC inhibited the receptor expression in these cells. These data suggested that in JAR cells, The EGF, E2 and DEX upregulated the PTH/PTHrP receptor expression, whereas the 1,25 DHCC down-regulated the PTH/PTHrP receptor, and the 1,25 DHCC may play an important role as antiproliferative drug for choriocarcinoma

Deflazacort Increases Osteoclast Formation in Mouse Bone Marrow Culture and the Ratio of RANKL/OPG mRNA Expression in Marrow Stromal Cells

Information on precise effects of deflazacort on bone cell function, especially osteoclasts, is quite limited. Therefore, the present study was undertaken to test effects of deflazacort on osteoclast-like cell formation in mouse bone marrow cultures and on the regulation of osteoprotegerin (OPG) and its ligand (RANKL) mRNA expressions by RT-PCR in the ST2 marrow stromal cells. TRAP-positive mononuclear cells increased after the treatment of deflazacort at 10(-9) to 10(-7) M alone for 6 days in a dose-dependent manner. Number of TRAP-positive multi-nucleated cells (MNCs) increased significantly with combined treatment of deflazacort at 10(-7) M and 1,25-(OH)2D3 at 10(-9) M compared to that of cultures treated with 1,25-(OH)2D3 alone (p<0.05). Exposure to deflazacort at 10(-7) M in the presence of 1,25-(OH)2D3 at 10(-9) M in the last 3-day culture had greater stimulatory effect on osteoclast-like cell formation than that of the first 3-day culture did. Deflazacort at 10(-10) -10(-6) M downregulated OPG and upregulated RANKL in mRNA levels in a dose-dependent manner. These observations suggest that deflazacort stimulate osteoclast precursor in the absence of 1,25-(OH)2D3 and enhance differentiation of osteoclasts in the presence of 1,25-(OH)2D3. These effects are, in part, thought to be mediated by the regulation of the expression of OPG and RANKL mRNA in marrow stromal cells.

La vitamina D induce la proliferación de células en cultivo del endometrio de rata / Vitamin D induces proliferation in rat endometrium cultured cells

Objetivo. Estudiar el efecto de 1,25-dihydroxycholecalciferol D(1,25-(OH)2D3 sobre la proliferación y muerte de las células de endometrio de la rata en cultivo. Material y métodos. Se usó la línea celular de endometrio de rata Rentro 1. El medio de incubación se suplementó con 1 por ciento de suero bovino fetal inactivado y previamente tratado con carbón para eliminar las hormonas esteroides. Las monocapas de células fueron mantenidas en presencia o ausencia de 1,25-(OH)2D3 o 17ß-estradiol o del vehículo. Posteriormente se evaluó la proliferación celular mediante conteo en un hemocitómetro, utilizando azul tripano 0.4 por ciento y se analizó la fase de síntesis de ADN por citofluorometría fe flujo. La muerte celular fue determinada por el análisis de la integridad del ADN genómico en geles de agarosa y tinción con bromuro de etidio. Resultados. Las células en presencia de 1 por ciento de suero bovina fetal sin hormonas esteroides en el medio de cultivo, estimuló su crecimiento de las mismas. Por otro lado, las células Rentro 1 no respondieron a la estimulación con 17ß-estradiol y sí al 1,25-(OH)2D3, lo que confirmó la ausencia del receptor de estrógenos en estas células y demostró la capacidad de esta línea celular para responder al 1,25-(OH)2D3. Por último, se encontró que a diferencia de otros tipos celulares, las células Rentro 1 no sufrieron daño a nivel del ADN (apoptosis) con el 1,25-(OH)2D3. Conclusiones. 1) El 1,25-(OH)2D3 promovió la proliferación de las células Rentro 1 de manera independiente de la dosis e independiente de la presencia del estímulo estrogénico; 2) el incremento en el número de células estuvo en relación con la activación de la fase de síntesis de ADN del ciclo celular; 3) la presencia de esta hormona en el cultivo celular no indujo la muerte celular no indujo la muerte celular por apoptosis

1, 25-Dihydroxyvitamin D3 stimulates growth and inhibits calcitonin secretion in a human C cell carcinoma cell line

1,25-Dihydroxyvitamin D3 (1,25D3), calcitonin (CT) and parathyroid hormone are the major calcium-regulating hormones. In addition, 1,25D3 has been reported to be a modulator of cell growth and differentiation in many tissues. recently, a suppressive effect of 1,25D3 on CT secretion and synthesis in C cells was demonstrated in vivo and also in vitro, but there are no data about in effects on thyroid C cell growth. We investigated the effects of [3H]-1,25D3 on basal and stimulated CT secretion and on [3H]-thymidine incorporation, using a human medullary thyroid carcinoma cell line (TT cells). After a 4-day expossure to 1,25D3, TT cells showed a dose-dependent inhibition of basal CT secretion (64 per cento of value for the control group at 100 nM 1,25D3). calcium (3mM) plus K+ (50mM) greatly increased CT secretion in both the control and vitamin D-treated groups. However, in the cells preincubated with 1,25D3 the stimulated CT levels less than observed in controls. A dose-dependent increase in [3H]-thymidine incorporation (200 per cent of the value for the at 100 nM 1,25D3) and in cell number (150 per cent of the value for the control group at 100 nM 1,25D3 after 72h) was observed in the groups treated with 1,25D3. 24,25D3 had no effect on CT secretion or cell growth compared to the control group. These data show that 1,25D3 decreased basal and Ca2+-stimulated CT secretion, a specialized function of these cells, and stimulated their growth. Hence, in contrast to its effects on other cell lines, 1,25D3 appears to induce a dedifferentiation on TT cell

Calcipotriol no tratamento da psoríase / Treatment of psoriasis by calcipotriol

O calcipotriol é um novo derivado sintético da vitamina D3 com marcada atuaçäo sobre a proliferaçäo e diferenciaçäo celular com efeito desprezível no metabolismo do cálcio. Em estudo aberto, näo comparativo, 21 pacientes com psoríase vulgar foram tratados com calcipotriol em pomada, duas vezes ao dia por seis semanas. Os pacientes foram avaliados e fotos tiradas nas semanas 0,2 e 6. O método de avaliaçäo foi o PASI (Psoriatic Area Severity Index) e revelou ao final das seis semanas uma reduçäo de 81% na descamaçäo. 82,2% na infiltraçäo e 57% no eritema. Nenhuma alteraçäo laboratorial foi detectada nos exames colhidos no momento da inclusäo e ao final do tratamento. Efeitos colaterais tais como prurido em quatro casos e ardência em um näo foram fortes o suficiente para interrupçäo do tratamento. Os resultados obtidos permitem-nos concluir que o calcipotriol constitui um medicamento bastante eficaz no tratamento da psoríase vulgar, sem efeito rebote, sem transtornos cosméticos e com poucos efeitos colaterais

Calcipotriol: una breve revisión / Calcipotriol: an short review

El ungüento de calcipotriol (50 mcg/g) es un tratamiento eficaz y seguro para la psoriasis. Probablemente actúa por regulación directa de los queratinocitos, inhibiendo la proliferación y aumentando su diferenciación. Debe ser aplicado dos veces al día de tal manera que menos de 100 g a la semana sean usados. Es tan eficaz como la betametasona y la antralina y es preferido por los pacientes. La irritación lesional y perilesional es común, pero los efectos adversos serios son raros. El calcipotriol ha sido usado en forma exitosa asociado con UVB, PUVA y ciclosporina en estudios preliminares. El calcio sérico, o la excresión urinaria de calcio, debe monitorizarse cuidadosamente si, a) más de 100g de calcipotriol se utilizan semanalmente, y b) el paciente presenta anomalías en el metabolismo del calcio o insuficiencia renal. Calcipotriol será usado en forma creciente en psoriasis y otras patologías cutáneas. Su lugar en la práctica dermatológica está asegurado y promete más en el futuro

Adverse effect of 1, 25-dihydroxycholecalciferol on erythrocyte precursors in the rats

La administración oral de 0.5 ugm/d/100 gm de peso a ratas normales produjo una significativa despoblación de la médula ósea, especialmente en la línea eritroide. El análisis de muestras de sangre periférica mostró ligera anemia con reticulocitopenia. La tasa de desaparición de eritrocitos marcados con 51-cr no fue afectada por el tratamiento, lo que sugiere que éste impide que la médula ósea reponga los eritrocitos a la tasa normal. El efecto adverso del tratamiento sobre la eritropoyesis parece relacionado con el grado de hipercalcemia que, a su vez, depende del contenido de la dieta