cholecalciferol and Fibrosis

Transforming growth factor-beta (TGF-beta) plays a central role in fibrosis, contributing to the influx and activation of inflammatory cells, the epithelial to mesenchymal transdifferentiation (EMT) of cells and the influx of fibroblasts and their subsequent elaboration of extracellular matrix. TGF-beta signals through transmembrane receptor serine/threonine kinases to activate novel signalling intermediates called Smad proteins, which modulate the transcription of target genes. The use of mice with a targeted deletion of Smad3, one of the two homologous proteins which signals from TGF-beta/activin, shows that most of the pro-fibrotic activities of TGF-beta are mediated by Smad3. Smad3 null inflammatory cells and fibroblasts do not respond to the chemotactic effects of TGF-beta and do not autoinduce TGF-beta. The loss of Smad3 also interferes with TGF-beta-mediated induction of EMT and genes for collagens, plasminogen activator inhibitor-1 and the tissue inhibitor of metalloprotease-1. Smad3 null mice are resistant to radiation-induced cutaneous fibrosis, bleomycin-induced pulmonary fibrosis, carbon tetrachloride-induced hepatic fibrosis as well as glomerular fibrosis induced by induction of type 1 diabetes with streptozotocin. In fibrotic conditions that are induced by EMT, such as proliferative vitreoretinopathy, ocular capsule injury and glomerulosclerosis resulting from unilateral ureteral obstruction, Smad3 null mice also show an abrogated fibrotic response. Animal models of scleroderma, cystic fibrosis and cirrhosis implicate involvement of Smad3 in the observed fibrosis. Additionally, inhibition of Smad3 by overexpression of the inhibitory Smad7 protein or by treatment with the small molecule, halofuginone, dramatically reduces responses in animal models of kidney, lung, liver and radiation-induced fibrosis. Small moleucule inhibitors of Smad3 may have tremendous clinical potential in the treatment of pathological fibrotic diseases.

Topics: Animals; Cholecalciferol; Cytokines; DNA-Binding Proteins; Extracellular Matrix; Fibrosis; Gene Expression Regulation; Humans; Signal Transduction; Smad2 Protein; Smad3 Protein; Smad7 Protein; Trans-Activators; Transforming Growth Factor beta; Wound Healing

Topics: Cholecalciferol; Cytokines; Fibroblast Growth Factor 2; Fibrosis; Humans; Interleukin-17; Leukocytes, Mononuclear; Scleroderma, Systemic; Transforming Growth Factor beta

Toll-like receptor 3 (TLR3) activation by viral infections plays a key role in promoting inflammatory immune responses that contribute to pulmonary fibrosis in chronic inflammatory respiratory diseases. Vitamin D3 has been shown to be beneficial to patients with asthma and chronic obstructive pulmonary disease (COPD) through its anti-inflammatory and anti-fibrotic properties. Smooth muscle cells are one of the major contributors to airway remodeling in asthma and COPD. We therefore aimed to investigate the effect of vitamin D3 treatment on viral-induced TLR3 responses in Bronchial Smooth Muscle Cells (BSMCs) as a mechanism contributing to pulmonary fibrosis in asthma and COPD. Primary BSMCs from patients with asthma (n=4), COPD (n=4), and healthy control subjects (n=6) were treated with polyinosinic: polycytidylic acid (polyI:C), TLR3 agonist in the presence or absence of vitamin D3 (1,25D3). Here we report the mRNA expression and protein levels of pro-inflammatory and pro-fibrotic markers (IL-6, IFN-β1, CCL2/MCP-1, fibronectin 1 and type I collagen) among BSMCs groups: asthma, COPD, and healthy controls. We show that at the baseline, prior to polyI:C stimulation, asthma and COPD BSMCs presented increased pro-inflammatory and pro-fibrotic state compared to healthy control subjects, as measured by quantitative PCR and immunoassays (ELISA/Flow Cytometry. Ligation of TLR3 by polyI:C in BSMCs was associated with increased

Topics: Adult; Asthma; Biomarkers; Bronchitis; Cells, Cultured; Cholecalciferol; Cytokines; Disease Susceptibility; Female; Fibrosis; Gene Expression Regulation; Humans; Inflammation Mediators; Male; Middle Aged; Myocytes, Smooth Muscle; Receptors, Calcitriol; Respiratory Function Tests; Vitamin D3 24-Hydroxylase; Young Adult

Renal fibrosis is common especially in the elderly population. Recently, we found that vitamin D deficiency caused prostatic hyperplasia. This study aimed to investigate whether vitamin D deficiency promotes renal fibrosis and functional impairment. All mice except controls were fed with vitamin D-deficient (VDD) diets, beginning from their early life. The absolute and relative kidney weights on postnatal week 20 were decreased in VDD diet-fed male pups but not in female pups. A mild pathological damage was observed in VDD diet-fed male pups but not in females. Further analysis showed that VDD-induced pathological damage was aggravated, accompanied by renal dysfunction in 40-week-old male pups. An obvious collagen deposition was observed in VDD diet-fed 40-week-old male pups. Moreover, renal α-smooth muscle actin (α-SMA), a marker of epithelial-mesenchymal transition (EMT), and Tgf-β mRNA were up-regulated. The in vitro experiment showed that 1,25-dihydroxyvitamin D3 alleviated transforming growth factor-β1 (TGF-β1)-mediated down-regulation of E-cadherin and inhibited TGF-β1-evoked up-regulation of N-cadherin, vimentin and α-SMA in renal epithelial HK-2 cells. Moreover, 1,25-dihydroxyvitamin D3 suppressed TGF-β1-evoked Smad2/3 phosphorylation in HK-2 cells. These results provide experimental evidence that long-term vitamin D deficiency promotes renal fibrosis and functional impairment, at least partially, through aggravating TGF-β/Smad2/3-mediated EMT in middle-aged male mice.

Topics: Actins; Animals; Antigens, CD; Cadherins; Calcitriol; Cell Line; Cholecalciferol; Epithelial-Mesenchymal Transition; Female; Fibrosis; Humans; Kidney; Kidney Diseases; Kidney Tubules, Proximal; Male; Mice; Mice, Inbred ICR; Organ Size; Transforming Growth Factor beta; Vimentin; Vitamin D; Vitamin D Deficiency

Vitamin D. After 12 weeks, the vitamin D

Topics: Animals; Cholecalciferol; Diabetes Mellitus, Experimental; Dietary Supplements; Fibrosis; Gene Expression Regulation; Male; PPAR gamma; Rats; Rats, Sprague-Dawley; Spermatogenesis; Testis; Transforming Growth Factor beta1; Vitamins

Duchenne muscular dystrophy (DMD) is caused by a lack of dystrophin protein. Next to direct effects on the muscles, this has also metabolic consequences. The influence of nutrition on disease progression becomes more and more recognized. Protein intake by DMD patients may be insufficient to meet the increased demand of the constantly regenerating muscle fibers. This led to the hypothesis that improving protein uptake by the muscles could have therapeutic effects. The present study examined the effects of a modified diet, which composition might stimulate muscle growth, on disease pathology in the D2-mdx mouse model. D2-mdx males were fed with either a control diet or modified diet, containing high amounts of branched-chain amino acids, vitamin D3 and ursolic acid, for six weeks. Our study indicates that the modified diet could not ameliorate the muscle pathology. No effects on bodyweight or weight of individual muscles were observed. Neither did the diet affect severity of fibrosis or calcification of the muscles.

Topics: Amino Acids, Branched-Chain; Animals; Calcinosis; Cholecalciferol; Dietary Proteins; Disease Models, Animal; Dystrophin; Fibrosis; Humans; Male; Mice; Mice, Inbred DBA; Mice, Inbred mdx; Muscle, Skeletal; Muscular Dystrophy, Duchenne; Regeneration; Triterpenes; Ursolic Acid

To study the molecular effects of three different D-vitamins, vitamin D2, vitamin D3 and calcipotriol, in pancreatic stellate cells (PSCs).. Quiescent PSCs were isolated from mouse pancreas and activated. Sustained culture of originally quiescent PSCs induced cell proliferation, loss of lipid droplets and exhibition of stress fibers, indicating cell activation. When added to PSCs in primary culture, all three D-vitamins diminished expression of α-SMA (to 32%-39% of the level of control cells;. The three D-vitamins inhibit, with similar efficiencies, activation of PSCs

Topics: Actins; Animals; Calcitriol; Cell Proliferation; Cells, Cultured; Cellular Senescence; Cholecalciferol; DNA Replication; Ergocalciferols; Fibrosis; Interleukin-6; Lipid Droplets; Mice, Inbred C57BL; Myofibroblasts; Pancreas; Pancreatic Diseases; Pancreatic Stellate Cells; Phenotype; Proline

There was evidence that 1,25(OH)

Topics: Actins; Animals; Aquaporin 2; Aquaporin 4; Calcitriol; Cholecalciferol; Cholesterol; Disease Models, Animal; Down-Regulation; Emulsions; Epithelial Cells; Fibronectins; Fibrosis; Kidney; Male; Mice, Inbred ICR; Necrosis; Plasminogen Activator Inhibitor 1; Receptors, Calcitriol; Signal Transduction; Tissue Culture Techniques; Transforming Growth Factor beta; Ureteral Obstruction

BACKGROUND The aim of this study was to study the effects of 1-alpha,25-dihydroxy-cholecalcifero (1,25(OH)2D3) on liver fibrosis and the generation of Th17 cells in vivo and in vitro. MATERIAL AND METHODS Thirty C57 mice were randomly divided into control, model, and treatment groups. Hepatic fibrosis was induced by subcutaneous injection of CCl4. Liver fibrosis condition was evaluated through pathological inspection and blood biochemical examination of liver function. Immunohistochemical assays were used to detect the expression of α-SMA, TGF-β, and collagen I to observe hepatic stellate cell activation level. Flow cytometry, ELISA, and RT-PCR were performed to explore the association between 1,25(OH)2D3 and Th17 cell differentiation. RESULTS Collagen I, TGF-β, and α-SMA were decreased after 1,25(OH)2D3 treatment. Consistently, RORγt mRNA and the rate of Th17 cells was significantly reduced after 1,25(OH)2D3 treatment. In vitro, the proportion of Th17 cells was also obviously reduced in the 1,25(OH)2D3 group, and mRNA levels of IL-17A, IL-22, RORγt, and RORa were significantly decrease in the 1,25(OH)2D3 group compared to the control group. CONCLUSIONS Treatment with 1,25(OH)2D3 can alleviate the damage caused by liver fibrosis. Experiments in vivo and in vitro showed that 1,25(OH)2D3 treatment deceased the rates of Th1 and Th17 cells and increased the rate of Th2 cells. The level of IL-17A, IL-22 and IFN-γ were decreased, while the level of IL-4 was increased by the treatment of 1,25(OH)2D3.

Topics: Animals; Calcitriol; Cholecalciferol; Fibrosis; Hepatic Stellate Cells; Interleukin-17; Interleukin-22; Interleukins; Liver; Liver Cirrhosis; Mice; Mice, Inbred C57BL; Protective Agents; Th17 Cells; Th2 Cells

The recent discovery of the vitamin D receptor (VDR) in regenerating muscle raises the question regarding the action of vitamin D3 on skeletal muscle regeneration. To investigate the action of vitamin D3 on this process, the tibialis anterior muscle of male C57BL/6 mice (10 wk of age) was injected with 1.2% BaCl2 to induce extensive muscle injury. The bioactive form of vitamin D3 [1α,25(OH)2D3] was administered daily via intramuscular injections during the regenerative phase (days 4-7 postinjury). Physiological and supraphysiological doses of 1α,25(OH)2D3 relative to 1 μg/kg muscle wet weight and mouse body weight were investigated. Muscle samples were collected on day 8 postinjury to examine proteins related to vitamin D3 metabolism (VDR, CYP24A1, and CYP27B1), satellite cell differentiation and regenerative muscle fiber formation [myogenin and embryonic myosin heavy chain (EbMHC)], protein synthesis signaling (Akt, p70 S6K1, 4E-BP1, and myostatin), fiber-type composition (fast and slow MHCs), fibrous formation (vimentin), and angiogenesis (CD31). Administration of 1α,25(OH)2D3 at physiological and supraphysiological doses enhanced VDR expression in regenerative muscle. Moreover, CYP24A1 and vimentin expression was increased, accompanying decreased myogenin and EbMHC expression at the supraphysiological dose. However, there was no change in CYP27B1, Akt, p70 S6K1, 4E-BP1, myostatin, fast and slow MHCs, or CD31 expression at any dose investigated. Taken together, administration of 1α,25(OH)2D3 at a supraphysiological dose decreased satellite cell differentiation, delayed regenerative muscle fiber formation, and increased muscular fibrosis. However, protein synthesis signaling, fiber-type composition, and angiogenesis were not affected by either 1α,25(OH)2D3 administration at a physiological or supraphysiological dose.

Topics: Animals; Cell Differentiation; Cholecalciferol; Fibrosis; Injections, Intramuscular; Male; Mice; Mice, Inbred C57BL; Morphogenesis; Muscle, Skeletal; Muscular Diseases; Myogenin; Myosin Heavy Chains; Neovascularization, Pathologic; Receptors, Calcitriol; Regeneration; Wound Healing

Aldosterone (Aldo) is an important active hormone in the renin-angiotensin-aldosterone system and plays a vital role in the development of hypertension, heart failure and other cardiovascular diseases. We aimed to explore the role of endogenous Aldo in aortic calcification in rats. We induced arterial calcification in rats by intramuscular administration of vitamin D(3) plus oral nicotine (VDN) and determined calcium content, (45)Ca(2+) accumulation and activity of alkaline phosphatase (ALP). The mRNA level of osteopontin (OPN) was measured by semi-quantitative reverse transcriptase polymerase chain reaction. Deposition of collagen in the aorta wall was measured by Sirius red staining. The content of angiotensin II (Ang II) and Aldo in plasma and myocardial and vascular tissue was determined by radioimmunoassay. In rats with VDN treatment, von Kossa staining showed calcification in vascular smooth muscle cells and extracellular matrix, and the content of calcium in calcified arteries was 5.8-fold of that in control arteries (P < 0.01). The accumulation of (45)Ca(2+) and activity of ALP in calcified aortic tissue was three- and 2.5-fold, respectively, that in control tissue (P < 0.01). The mRNA expression of OPN was significantly higher, by 58%, in calcified than control tissue (P < 0.01). Vascular fibrosis was greater in rats with calcified tissue than in control rats. The level of Ang II and Aldo was 58% and 80% higher, respectively, in calcified than control tissue (both P < 0.01). The changes could be significantly improved by treatment with captopril, an angiotensin-converting enzyme inhibitor, and the Aldo receptor antagonist spironolactone. These results suggest that Aldo is an endogenous bioactive factor involved in vascular calcification.

Topics: Aldosterone; Alkaline Phosphatase; Angiotensin II; Angiotensin-Converting Enzyme Inhibitors; Animals; Aorta; Calcium; Captopril; Cholecalciferol; Collagen; Fibrosis; Hypertension; Male; Mineralocorticoid Receptor Antagonists; Nicotine; Osteopontin; Rats; Rats, Sprague-Dawley; RNA, Messenger; Spironolactone; Vascular Calcification

Topics: Animals; Bone Density; Calcium; Cells, Cultured; Cholecalciferol; Dermis; Female; Fibroblast Growth Factor-23; Fibroblast Growth Factors; Fibroblasts; Fibrosis; Humans; Mice; Mice, Inbred C57BL; Phosphates; Vitamin D