Role of vitamin D3 combined to alginates in preventing acid and oxidative injury in cultured gastric epithelial cells

Abstract

BackgroundGastric diseases are a worldwide problem in modern society, as reported in the USA, in the range of 0.5–2 episodes/year/person and an incidence of 5–100 episodes/1000/week according to seasons and age. There is convincing evidence that oxidative stress is involved in the pathogenesis of acute gastric injury. Acid secreted from gastric parietal cells determines mucosal injuries which in turn cause inflammation and oxidative stress. Consequent inflammation produces free radicals by mitochondria thus causing lipid peroxidation, oxidative and acidic stress, which can lead to cell apoptosis. Vitamin D3, the active form of vitamin D, may counteract intracellular cell death and improve epithelial regeneration.MethodsThis study was planned to assess whether vitamin D3 is a protective factor against acid injury and oxidative stress in gastric epithelial cells. Primary epithelial cells and GTL-16 cells have been used to test the effects of Grisù® alone or in combination with vitamin D3 during oxidative stress or high acid exposition measuring cell viability, ROS production, cellular adhesion time along with apoptotic, autophagic and survival pathways. The combined effect of Grisù® and vitamin D3 was found more effective in counteracting the negative consequences of oxidative stress and acidity conditions than some other gastroprotective agents, such as Maalox® or Gaviscon®.ResultsIn case of oxidative stress or acidity condition the stimulation with Grisù® alone caused an improvement of cell viability and a reduction of ROS production on epithelial gastric cells. In addition, the adhesion time of the cells was improved. All these effects were increased by the presence of vitamin D3. Similar data were also observed in primary gastric epithelial cells confirming the results obtained in GTL-16 cells.ConclusionsThese results suggest that Grisù® in combination with vitamin D3 may exert a gastroprotective effect to maintain or restore the integrity of gastric epithelium through an antioxidant pathway, inhibiting apoptosis and activating survival kinases. Moreover, the combination of Grisù® and vitamin D3 improves cell viability and decreases ROS production compared to other gastroprotective agents combined with vitamin D3. All these data were validated using primary cells isolated from gastric tissue.Electronic supplementary materialThe online version of this article (doi:10.1186/s12876-016-0543-z) contains supplementary material, which is available to authorized users.

Vitamin D Receptor Is Necessary for Mitochondrial Function and Cell Health

“…In line with this, we demonstrated that, after two weeks of absence of VDR, mitochondrial integrity was lost, and the cells showed the signs of an apoptotic fate. The interplay between vitamin D–VDR, ROS signaling, and the antioxidant system is complex; on the one hand, it has been demonstrated that vitamin D and its analogues can increase the cytotoxicity mediated by ROS [ 36 ], while, on the other hand, few reports have proved that vitamin D–VDR is able to inhibit the apoptosis triggered by oxidative stress [ 37 , 38 , 39 , 40 , 41 ]; in addition, in this study, for the first time, we showed the protective role exerted by VDR itself, without additional stressors. Moreover, our study investigated a novel mechanism involved in the antiapoptotic effects of VDR, previously ascribed only to its transcriptional activity [ 42 ].…”

Biological effects of combined resveratrol and vitamin D3 on ovarian tissue

“…MTT-based In Vitro Toxicology Assay Kit (Sigma-Aldrich) was performed as described in literature [ 40 ] to determine cell viability after stimulations. Cells were incubated in DMEM without red phenol 0% FBS with 1% MTT dye for 2 h at 37 °C in incubator [ 41 ] and then cell viability was determined measuring the absorbance through a spectrometer (VICTORX4 multilabel plate reader) at 570 nm with correction at 690 nm.…”

Iron Absorption from Three Commercially Available Supplements in Gastrointestinal Cell Lines

“…GTL-16 cell line, donated by the Laboratory of Histology of the University of Eastern Piedmont, is a clonal line derived from a poorly differentiated gastric carcinoma cell line [ 10 ] widely used as a model of gastric epithelial cells. Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 1% penicillin-streptomycin in incubator at 37 °C, 5% CO 2 [ 11 ]. This cell line was plated at different densities, 1 × 10 4 cells were plated on 96-well plates to study cell viability (MTT test); 2 × 10 4 cells were seeded onto 6.5 mm translucent polyethylene terephthalate (PET) transwell insert 0.4 μm in a 24 well to study absorption; 0.5 × 10 4 cells were placed in Culture Slide (BD Biosciences, Bedford, MA, USA) with 4 chambers to perform immunohistochemistry tests.…”

“…The cells plated on transwell insert were maintained in complete medium changed every other day, first basolaterally and then apically for 7 days before the stimulations. Before stimulations, cells were synchronized by incubation in DMEM without red phenol and FBS and supplemented with 1% penicillin/streptomycin, 2 mM l -glutamine and 1 mM sodium pyruvate in an incubator at 37 °C, 5% CO 2 , and 95% humidity for 18 h [ 11 ].…”

“…MTT-based In Vitro Toxicology Assay Kit (Sigma-Aldrich) was performed as described in literature [ 11 ] to determine cell viability after stimulations. Briefly, at the end of treatments, Caco-2 and GTL-16 cells were incubated with 1% MTT dye in DMEM without red phenol 0% FBS for 2 h at 37 °C in incubator [ 13 ] and cell viability was determined measuring absorbance at 570 nm with correction at 690 nm through a spectrometer (VICTORX4 multilabel plate reader, PerkinElmer, Waltham, MA, USA).…”