Biomedical subjects

A Sellmayer

Publications and source records attributed to A Sellmayer.

At least 19 recordsLinked to original sources

PubMed1999

Lipids in vascular function.

Physiological and pathological vascular responses depend on the action of numerous intercellular mediators, ranging from hormones to gases like nitric oxide, proteins, and lipids. The last group consists not only of the different types of lipoproteins, but also includes a broad array of other lipophilic signaling molecules such as fatty acids, eicosanoids, phospholipids and their derivatives, sphingolipids and isoprenoids. Due to space limitations, it is impossible to discuss all the vascular effects of lipophilic mediators or compounds. Therefore, we will focus on one of the most important lipid-mediated diseases, atherosclerosis. Lipoproteins and especially their native or oxidized lipid compounds affect vascular function in many different ways, and these effects do not only modulate atherogenesis but are of paramount physiological and pathophysiological importance in other diseases, such as inflammation, tumor metastasis, or normal wound healing.

Animals↗

PubMed1999

Long-chain polyunsaturated fatty acids and eicosanoids in infants--physiological and pathophysiological aspects and open questions.

Eicosanoids are highly active lipid mediators in physiologic and pathologic processes, with their effects ranging from cytoprotection and vasoactivity to modulation of inflammatory and proliferative reactions. Generation of eicosanoids can be affected by changes in the pools of their precursors, the long-chain polyunsaturated fatty acids (LCPUFA). Thus, dietary interventions such as supplementation of infant formula with specific n-3 and n-6 LCPUFA will alter formation as well as activity of the eicosanoids produced. This report summarizes the results and discussion of the workshop on "Eicosanoids and Polyunsaturated Fatty Acids in Infants." The intention of the workshop organizers was to give an overview of the role of eicosanoids in physiological and pathophysiological processes in infants, to discuss the implications that an increased n-3 and n-6 LCPUFA intake may have on eicosanoid generation, and to point out open questions and controversies for future research.

Central Nervous System↗

PubMed1998

High glucose increases prostaglandin E2 synthesis in human peritoneal mesothelial cells: role of hyperosmolarity.

Peritoneal mesothelial cells are considered the predominant source of peritoneal prostanoid formation because they represent the largest resident cell population in the peritoneal cavity. The present study was designed to evaluate the effect of D-glucose, which is widely used in commercially available peritoneal dialysis fluids as an osmotic compound, on the synthesis of prostaglandins in cultured human mesothelial cells (HMC). Analysis of eicosanoid synthesis in HMC by reversed-phase HPLC revealed that 6-keto-PGF1alpha, the spontaneous hydrolysis product of prostacyclin (PGI2), and prostaglandin E2 (PGE2) were the main eicosanoids produced. Addition of D-glucose resulted in a time- and concentration-dependent (30 to 120 mM) increase in PGE2 production in HMC (24 h, 90 mM: 3.9+/-0.5 ng/10(5) cells versus 2.3+/-0.3 in untreated cells; P < 0.05). Mannitol (90 mM) or L-glucose (90 mM). nonmetabolizable osmotic compounds, also led to a significant (P < 0.05) but less intense increase in PGE2 synthesis (3.3+/-0.4 and 3.2+/-0.5 ng/10(5) cells, respectively). Increased PGE2 synthesis was completely blunted by coincubation with the specific protein kinase C (PKC) inhibitor Ro 31-8220 or downregulation of PKC activity by preincubation with phorbol myristate acetate for 16 h. Furthermore, coincubation with PD 98059, an inhibitor of the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway, also inhibited increased PGE2 synthesis by D-glucose or mannitol. In contrast, the iso-osmolar glucose polymer icodextrin, which is used as an alternative to D-glucose in peritoneal dialysis solutions, had no effect on PGE2 synthesis. These data indicate that D-glucose and metabolically inert sugars increase PGE2 synthesis in HMC at least in part by hyperosmolarity and that this effect requires activation of PKC and the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway of intracellular signaling.

Arachidonic Acid↗

PubMed1997

Intrinsic cyclooxygenase activity is not required for monocytic differentiation of U937 cells.

Metabolism of arachidonic acid has been found to modulate cell growth and differentiation. Since differentiation of premonocytic U937 cells is paralleled by increase expression of cyclooxygenase and prostanoid synthesis, we assessed the role of cyclooxygenase in the differentiation of U937 cells. Treatment with retinoic acid (RA, 1 microM) or 1,250(OH)2-vitamin D3 (1,25-D3, 10 nM) increased the expression of the monocytic surface antigens CD11b plus CD11c, and CD11b, CD11c, CD14 plus CD18, respectively. Addition of the cyclooxygenase inhibitors indomethacin (10 microM) or piroxicam (20 microM) slightly increased expression of CD11b and CD18 in cells differentiated with RA, but did not alter expression of surface antigens in cells treated with 1,25-D3. Stimulus-dependent rises in the cytosolic-free calcium concentrations remained unchanged by the inhibitors, as was superoxide anion generation in cells stimulated with phorbol myristate acetate. Effective inhibition of cyclooxygenase over the 72 h of differentiation was proven by the marked decrease in A23187-stimulated prostanoid formation in differentiated cells. To assess whether prostaglandins negatively control monocytic differentiation, as suggested by the stimulatory effects of cyclooxygenase inhibitors on CD11b/CD18 expression in RA-treated cells, prostaglandin E2 (PGE2; 100 nM) was added to the differentiation with RA or 1,25-D3. However, the addition of PGE2 increased expression of CD11b and CD11b plus CD14, as well as superoxide anion generation. These data indicate that intrinsic cyclooxygenase activity is not required for monocytic differentiation of U937 cells. In addition, basal prostanoid secretion does not measurably contribute to monocytic differentiation.

Antigens, CD↗

PubMed1997

Modulation of the expression of early genes by polyunsaturated fatty acids.

Expression of early genes is a characteristic immediate cellular response to mitogenic or inflammatory stimulation. Various second messenger systems have been found to transduce the signal from the plasma membrane to the nucleus. Recent observations indicate that in addition to well characterized second messenger systems, polyunsaturated fatty acids, especially the n-6 fatty acid arachidonic acid and its endogenously produced metabolites affect the expression of early genes in different cell types. At least in fibroblasts, the stimulatory effect of arachidonic acid can be antagonized by n-3 polyunsaturated fatty acids. Further identification of the mechanisms through which polyunsaturated fatty acids modulate early gene expression and regulate subsequent cellular responses, like cell growth, may help to define novel concepts in the management of cardiovascular diseases.

3T3 Cells↗

PubMed1997

Hemodialysis-induced upregulation of granulocyte adhesion molecules CD11b/CD18 is independent from 5-lipoxygenase activity.

Hemodialysis with new cuprophane membranes upregulates expression of granulocyte adhesion molecules and activates 5-lipoxygenase as reflected by the enhanced generation of leukotriene B4 (LTB4). We assessed the role of 5-lipoxygenase activity in hemodialysis-induced upregulation of the granulocyte adhesion molecules, CD11b and CD18, and granulocyte adhesion to human umbilical vein endothelial cells in an ex vivo dialysis model. 5-Lipoxygenase was effectively inhibited by preincubation of human whole blood with the specific inhibitor, BAY X1005. Dialysis with new cuprophane but not with new acrylonitrile membranes significantly upregulated expression of CD11b and CD18 by 6-fold and 4-fold, respectively. Inhibition of 5-lipoxygenase did not affect the expression of CD11b and CD18 during dialysis with either of the two membranes. In contrast to the enhanced expression of CD11b and CD18, adhesion of granulocytes to human umbilical vein endothelial cells did not increase during dialysis, nor was it affected by BAY X1005. These data indicate that ex vivo dialysis with cuprophane membranes upregulates expression of CD11b and CD18 on granulocytes independent of the activation of 5-lipoxygenase.

Adult↗

PubMed1996

Linoleic acid esterified in low density lipoprotein serves as substrate for increased arachidonic acid synthesis in differentiating monocytic cells.

The cellular metabolism of albumin- and lipoprotein-bound 18:2(n - 6) during monocytic differentiation was examined in the human premonocytic U937 and Mono Mac 6 cells. Differentiation for 72 h of U937 cells with retinoic acid (RA, 1 microM) or 1,25-(OH)2-vitamin D3 (1,25-D3, 10 nM) and of Mono Mac 6 cells with RA (1 microM) or lipopolysaccharide (LPS, 10 ng/ml) increased the desaturation and elongation of [1-14C]18:2(n - 6) to [1-14C]20:4(n - 6). In undifferentiated U937 and Mono Mac 6 cells, incubations with human LDL (100 micrograms/ml, 18 h) resulted in a 2.5-fold increase in 18:2(n - 6) levels in the cellular phospholipids. Differentiation of U937 cells with RA or or of Mono Mac 6 cells with LPS prior to LDL addition. Significantly reduced 18:2(n - 6) and elevated 20:4(n - 6) levels in cellular phospholipids. This increase in 20:4(n - 6) was likely not due to an increased incorporation of preformed 20:4(n - 6) esterified in LDL, as the receptor-specific degradation of [125I]LDL was reduced in both the RA-treated U937 and LPS-treated Mono Mac 6 cells. In U937 cells incubated with [1-14C]18:2(n - 6), the synthesis of TXB2, PGE2 and HHT could be detected after differentiation with RA. suggesting the availability of [1-14C]20:4(n - 6), derived from [1-14C]18:2(n - 6), for cyclooxygenase metabolism. Our results show that the conversion of 18:2(n - 6) to 20:4(n - 6) increases during monocyte differentiation. The 18:2(n - 6) supplied to the cells via the receptor-mediated uptake of LDL was utilized as substrate for the increased 20:4(n - 6) synthesis.

Arachidonic Acid↗

PubMed1996

Differential effects of n-6 and n-3 polyunsaturated fatty acids on cell growth and early gene expression in Swiss 3T3 fibroblasts.

Dietary n-3 polyunsaturated fatty acids (PUFAs) have been found to reduce accelerated cell growth. To study the underlying molecular mechanisms, we evaluated the effects of the n-3 PUFAs eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) compared with the n-6 PUFA arachidonic acid (AA) on cell growth and early gene mRNA accumulation in Swiss 3T3 fibroblasts. AA significantly increased cell numbers and incorporation of [3H]-thymidine compared with cells treated with EPA and DHA, which did not stimulate cell growth. In contrast to AA and parallel to its effect on cell growth, EPA and DHA did not lead to a pronounced increase in Egr-1 and c-fosmRNA levels. When they were incubated together with AA, both DHA and EPA reduced AA-induced Egr-1 and c-fosmRNA accumulation and incorporation of [3H]-thymidine. We have recently shown that AA strongly increases Egr-1 and c-fosmRNA accumulation 3T3 fibroblasts through its metabolism to prostaglandin E2 (PGE2) and its subsequent activation of protein kinase C (Danesch et al., 1994, J. Biol. Chem., 269:27258-27263). Consistent with the notion that increased PGE2 formation is required for the AA-induced early gene mRNA accumulation, EPA and DHA reduced PGE2 formation from exogenous [14C]-AA by more than 60%, but they did not decrease mRNA levels following stimulation with PGE2. We suggest that, in 3T3 fibroblasts, EPA and DHA antagonize AA-induced early gene mRNA accumulation and cell growth by reducing PGE2 formation.

3T3 Cells↗

PubMed1996

Effects of different polyunsaturated fatty acids on growth-related early gene expression and cell growth.

Effects of polyunsaturated fatty acids on expression of early-response genes c-fos and Egr-1 and induction of cell growth were assessed in Swiss 3T3 fibroblasts. Stimulation with arachidonic acid increased mRNA levels of c-fos and Egr-1. This effect was inhibited by preincubation with cyclooxygenase inhibitors and restored by addition of prostaglandin E2 (PGE2), the predominant eicosanoid produced in Swiss 3T3 fibroblasts. Further signaling of PGE2, was mediated by a protein kinase C-dependent pathway, since downregulation, or inhibition, of protein kinase C reduced increases in mRNA levels. Parallel to the stimulatory effects on mRNA levels, AA and PGE2 also increased cell growth, as determined by uptake of [3H]-thymidine. In contrast to arachidonic acid, n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) did not increase c-fos and Egr-1 mRNA levels or cell growth. Furthermore, preliminary data indicate that EPA and DHA even reduce the stimulatory effect of AA, which is associated with reduced formation of PGE2. In conclusion, our data indicate that AA increases expression of growth-related early genes c-fos and Egr-1 Swiss 3T3 fibroblasts by its conversion to PGE2 and subsequent activation of protein kinase C, whereas n-3 fatty acids do not activate this signaling cascade.

3T3 Cells↗

PubMed1996

Arachidonic acid increases activation of NADPH oxidase in monocytic U937 cells by accelerated translocation of p47-phox and co-stimulation of protein kinase C.

Arachidonic acid (AA) has been implicated as an important amphiphilic co-factor in the activation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in neutrophils and reconstituted cell-free systems. To assess the role of AA in the activation of O2- generation in monocytic cells, we studied pre-monocytic U937 cells differentiated with 1,25-(OH)2-vitamin D3 plus interferon-gamma (IFN-gamma). AA dose-dependently enhanced phorbol myristate acetate (PMA)-stimulated O2- generation, with a maximum increase of 4,5-fold, through: (1) a more than 50% reduction of the lag-phase, defined as the time between addition of PMA and detection of O2-; and (2) a more than 60% increase in the constant rate of O2- generation. Reduction of the lag phase was associated with increased protein kinase C (PKC)-independent translocation of the cytosolic subunit of NADPH oxidase p47-phox to the cell membrane, whereas increased generation of O2- correlated with enhanced activation of PKC. The data indicate that AA increases activation of NADPH oxidase by accelerating its assembly and by co-stimulating PKC in monocytic U937 cells.

Arachidonic Acid↗

PubMed1996

[Nutrition in prevention of coronary heart disease].

Clinical as well as basic research in the field of atherogenesis indicates that the progression of this disease process can be slowed down or even reversed. It is well established that nutrition plays an important role in the prevention and treatment of the classical atherogenic risk factors such as obesity, diabetes mellitus and hyperlipidemia. In addition, some nutrients such as the polyunsaturated n-3-fatty-acids or antioxidative vitamins can intervene directly by influencing one or more steps of the atherogenetic and/or thrombogenetic process. A comprehensive understanding of the pathogenesis of this disease as well as of the mechanisms of nutrient action are essential to the planning of successful nutritional prevention strategies. Because most nutrients influence mainly the slow and long-standing development of the atherosclerotic lesion, their inclusion in primary nutritional prevention should be started at an early age. Few nutrients such as the n-3 fatty acids, which also reduce the thrombogenetic risk factors, have demonstrated some success in the secondary prevention of CHD. Given the complexity with which nutrients intervene in the atherosclerotic process and their interactions with each other, nutritional prevention strategies should be based on well-grounded dietary modifications rather than supplementation with individual nutrients.

Antioxidants↗

PubMed1995

Effects of dietary fish oil on ventricular premature complexes.

For ethical and practical reasons, in this study the antiarrhythmic potential of fish oil was evaluated in patients free from complex ventricular arrhythmias and severe heart failure. Although subjects without overt structural heart disease had ventricular arrhythmias that were not associated with an increased risk for sudden cardiac or coronary death, recent data suggest that frequent VPCs in patients similar to our study population may reflect subclinical cardiac disease amenable to the multiple beneficial actions of n-3 fatty acids. The potential and safety of fish oil as a treatment for more complex cardiac arrhythmias or arrhythmias in higher risk patients with more severe heart disease deserve further study.

Aged↗

PubMed1995

Cooperative effects of interferon-gamma on the induction of NADPH oxidase by retinoic acid or 1,25(OH)2-vitamin D3 in monocytic U937 cells.

The effect of retinoic acid (RA), 1,25-dihydroxyvitamin D3 (1,25-D3) or human recombinant interferon-gamma (IFN-gamma) on the induction of NADPH oxidase was studied in premonocytic U937 cells. Differentiation with the combination of either RA (1 microM) or 1,25-D3 (10 nM) with IFN-gamma (100 IU/ml) induced NADPH oxidase activity as demonstrated by increased superoxide anion (O2-) generation in response to stimulation with phorbol myristate acetate (PMA, 100 nM). Induction of NADPH oxidase activity was preceded by increases in mRNA levels of p47-phox, p67-phox and gp91-phox, which encode three subunits of the enzyme, and immunoblot analysis of the p47-phox and p67-phox proteins revealed that the increases in mRNA levels were equally reflected by increases in protein levels. In contrast, RA, 1,25-D3 or IFN-gamma alone did not induce NADPH oxidase activity which correlated with their failure to increase p67-phox and gp91-phox mRNA levels. The mRNA of p21 rac1, a GTP-binding protein that regulates NADPH oxidase activity in macrophages, was constitutively expressed in undifferentiated cells and was not affected by differentiation. These data indicate that induction of a functional NADPH oxidase in premonocytic U937 cells requires the cooperative actions of IFN-gamma plus RA or 1,25-D3 and is reflected in the increased expression of p67-phox and gp91-phox.

Calcitriol↗

PubMed1995

Differential effects of polyunsaturated fatty acids on cell growth and differentiation of premonocytic U937 cells.

The effect of long chain polyunsaturated fatty acids (PUFA) on cell growth and differentiation was assessed in human premonocytic U937 cells. Addition of either 10 microM arachidonic acid (AA, 20:4n-6), eicosapentaenoic acid (EPA, 20:5n-3) or docosahexaenoic acid (DHA, 22:6n-3) resulted in the rapid incorporation of these fatty acids into cellular phospholipids. Their uptake was greatest in the first 2 h. AA and EPA reached steady-state levels after 8 h, while levels of DHA increased steadily over 72 h. In parallel, fatty acid metabolites derived from AA and EPA, 22:4n-6, 22:5n-6 and 22:5n-3, 22:6n-3, respectively, increased continuously indicating an active fatty acid elongation and desaturation. The effects of PUFA on monocytic differentiation were examined in cells which had been enriched with AA, EPA or DHA for 8 h and subsequently treated with retinoic acid (RA), 1,25-(OH)2-vitamin D3 (1,25-D3), interferon-gamma (IFN-gamma) or their combinations for 72 h. Growth of differentiating or non-differentiating U937 cells was not affected by enrichment with PUFA. However, in cells differentiated with 1,25-D3 plus IFN-gamma, prior enrichment with all three PUFA slightly but significantly (P < 0.05) increased the expression of the monocytic surface antigens CD11b and CD14 and generation of superoxide anion. The data indicate that although n-6 and n-3 PUFA are rapidly incorporated into phospholipids, they do not affect cell growth. However, enrichment with PUFA increases monocytic differentiation of U937 cells when induced most effectively with 1,25-D3 plus IFN-gamma.

Antigens, CD↗

PubMed1994

Arachidonic acid increases c-fos and Egr-1 mRNA in 3T3 fibroblasts by formation of prostaglandin E2 and activation of protein kinase C.

Studying Swiss 3T3 fibroblasts, we report that arachidonic acid strongly stimulates mRNA levels of the growth-associated immediate early genes c-fos and Egr-1. Structurally related compounds like arachidonic acid methyl ester, arachidonyl alcohol, or eicosatetraynoic acid are ineffective, indicating a specific role of free unesterified arachidonic acid or an arachidonic acid metabolite in c-fos and Egr-1 mRNA accumulation. Blocking the conversion of arachidonic acid to prostaglandins by inhibiting cyclooxygenase abolishes arachidonic acid-induced accumulation of c-fos and Egr-1 mRNA. Inhibition of the lipoxygenase or cytochrome P-450 epoxygenase pathways has no significant effect on arachidonic acid-induced c-fos and Egr-1 mRNA levels, indicating that prostaglandin synthesis is necessary for the increase in c-fos and Egr-1 mRNA. Reversed phase high performance liquid chromatography revealed prostaglandin E2 (PGE2) as the major arachidonic acid metabolite in Swiss 3T3 fibroblasts. When added to the cells, PGE2 stimulates c-fos and Egr-1 mRNA levels to the same degree as arachidonic acid. Also, the inhibition of arachidonic acid-stimulated c-fos and Egr-1 mRNA accumulation by indomethacin is reversed by PGE2. Contrary to reports that PGE2 caused an increase in cAMP levels in Swiss 3T3 fibroblasts, we found that arachidonic acid and PGE2 only minimally increase cAMP levels as compared with untreated cells. In contrast, inhibition of protein kinase C by calphostin C and chelerythrine or down-regulation with phorbol 12-myristate 13-acetate drastically reduces PGE2 and arachidonic acid-induced c-fos and Egr-1 mRNA levels. These data indicate that arachidonic acid exerts its stimulatory effect on c-fos and Egr-1 mRNA via synthesis of PGE2 and subsequent activation of protein kinase C, probably through a PGE2 receptor coupled to phospholipase C.

3T3 Cells↗

PubMed1994

Differential induction of eicosanoid synthesis in monocytic cells treated with retinoic acid and 1,25-dihydroxy-vitamin D3.

Synthesis and release of eicosanoids is characteristic to mature monocytes, while in undifferentiated premonocytic cells arachidonic acid (AA) metabolism is barely detectable. Since some vitamins and cytokines induce differentiation of these cells to a more monocyte-like cell type, we evaluated the effect of retinoic acid (RA), 1,25(OH)2-vitamin D3 (1,25-D3) and interferon-gamma (IFN-gamma) on AA metabolism in the human premonocytic cell lines U937 and THP-1. In U937 cells, differentiation with RA (1 microM) followed by stimulation with the calcium ionophore A23187 (10 microM) or platelet activating factor (PAF; 100 nM) significantly increased synthesis of immunoreactive 6-keto PGF1 alpha, TxB2 and PGE2 5- to 6-fold. Analysis of AA metabolism in RA-differentiated cells prelabelled with [3H]-AA revealed the formation of additional radioactive compounds which coeluted with standards for PGF2 alpha, PGD2 and 12-hydroxyheptadecatrienoic acid (HHT). The structural identity of 6-keto PGF1 alpha, TxB2, PGE2 and HHT was confirmed by gas chromatography-mass spectrometry (GC-MS). In parallel, RA induced the expression of the monocytic surface antigen CD11b, but not CD14. Differentiation with 1,25-D3 (10 nM) only marginally increased stimulated eicosanoid formation, while it more effectively induced expression of CD11b and CD14. Pretreatment with IFN-gamma (100 IU/ml) slightly increased stimulus-dependent AA metabolism, but did not induce expression of CD11b or CD14. Induction of eicosanoid synthesis by RA was further confirmed in THP-1 cells. These data indicate that RA most effectively induced cyclooxygenase activity and stimulus-dependent eicosanoid formation in U937 and THP-1 cells. Furthermore, since expression of monocytic surface antigens differed between RA and 1,25-D3, it is suggested that induction of cyclooxygenase activity may correlate to differentiation into distinct monocytic phenotypes.

Antigens, Surface↗

PubMed1993

Platelet activating factor enhances receptor-operated Ca(++)-influx and subsequent prostacyclin synthesis in human endothelial cells.

Platelet activating factor (PAF) primes vascular actions of mediators such as histamine and stimulates human umbilical vein endothelial cells (HUVECs) to produce prostacyclin (PGI2), which is important for the regulation of vascular tone, perfusion and hemostasis. We demonstrate that pretreatment of HUVECs with PAF enhances thrombin- or histamine-induced rises of cytosolic free Ca(++)-concentration and subsequent Ca(++)-dependent PGI2-synthesis. Inhibition of Ca(++)-influx and PGI2-formation by SKF96365 (blocker of receptor-operated Ca(++)-channels) in PAF-treated cells indicates that the enhancement of PGI2-synthesis by PAF is due to sensitization of Ca(++)-entry. This suggests cooperative effects of PAF on activation processes induced by thrombin or histamine in HUVECs.

6-Ketoprostaglandin F1 alpha↗

PubMed1992

1 alpha,25-(OH)2 vitamin D3 enhances expression of the genes encoding Ca(2+)-binding proteins MRP-8 and MRP-14.

Two closely related Ca(2+)-binding proteins, migration inhibitory factor-related protein (MRP)-8 and MRP-14, are synthesized under specific conditions of myeloid cell differentiation. Because 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] induces myeloid cell differentiation and expression of other S-100 class calcium-binding proteins, we examined the effects of 1,25-(OH)2D3 on MRP mRNA levels in human U-937 histiocytic lymphoma cells. 1,25-(OH)2D3 increased MRP-8 and MRP-14 mRNA levels in a time- and dose-dependent manner. MRP mRNA levels were maximal at 24 h and remained elevated for at least 96 h after exposure of the cells to 1,25-(OH)2D3. MRP-8 mRNA accumulation required 100- to 1,000-fold higher concentrations of 25-(OH)D3, which binds to the 1,25-(OH)2D3 intracellular receptor with 100- to 1,000-fold lower affinity. Other differentiating agents, dimethyl sulfoxide, retinoic acid, and dexamethasone, also increased levels of MRP-8 and MRP-14 mRNA. Phorbol myristate acetate enhanced MRP-14 mRNA levels to a greater extent than MRP-8 mRNA levels, suggesting differential regulation of MRP gene expression by protein kinase C. The 1,25-(OH)2D3-induced relative increase in MRP mRNA levels was not changed by a 1,000-fold reduction in extracellular [Ca2+]. Thus 1,25-(OH)2D3 is potentially a physiological modulator of MRP gene expression. Expression of the MRP-8 and MRP-14 genes may be important for differentiation of myeloid cells.

Calcitriol↗