Influence of vitamin D on cell cycle, apoptosis, and some apoptosis related molecules in systemic lupus erythematosus

Tabasi, Nafise; Rastin, Maryam; Mahmoudi, Mahmoud; Ghoryani, Mohsen; Mirfeizi, Zahra; Zamani Taghizadeh Rabe, Shahrzad; Reihani, Hadi

doi:10.22038/ijbms.2015.6049

Influence of vitamin D on cell cycle, apoptosis, and some apoptosis related molecules in systemic lupus erythematosus

Document Type : Original Article

Authors

¹ Immunology Research Center, BuAli Research Institute, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

² Rheumatic Disease Research Center, Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran

10.22038/ijbms.2015.6049

Abstract

Objective(s):Genetic and environmental factors are involved in the pathogenesis of systemic lupus erythematosus (SLE). Autoreactive lymphocytes are cleared through apoptosis and any disturbance in the apoptosis or clearance of apoptotic cells may disturb tolerance and lead to autoimmunity. Vitamin D has anti-proliferative effects and controls cell cycle progression. In this study we investigated the effects of vitamin D on cell cycle and apoptosis induction in lupus patients.
Materials and Methods:Isolated peripheral blood mononuclear cells (PBMCs) from 25 SLE patients were cultured in the presence of 50 nM of 1,25(OH)2D3; then one part of the cells were stained with FITC labeled Annexin V and PI and were analyzed for apoptosis determination. For gene expression assessment of FasL, Bcl-2 and Bax, RNA was extracted from one another part of the cells, cDNA was synthesized and gene expression analysis was performed using Real time PCR. An additional part of the cells were treated with PI and the cell cycle was analyzed using flowcytometer.
Results: The mean number of early apoptotic cells in vitamin D treated cells decreased significantly (18.48±7.9%) compared to untreated cells (22.02±9.4%) (P=0.008). Cell cycle analysis showed a significant increase in G1 phase in vitamin D treated cells (67.33±5.2%) compared to non treated ones (60.77±5.7%) (P =0.02). Vitamin D up-regulated the expression levels of Bcl-2 by (18.87 fold increase), and down-regulated expression of Bax (23%) and FasL (25%).
Conclusion:Vitamin D has regulatory effects on cell cycle progression, apoptosis and apoptosis related molecules in lupus patients.

Keywords

References

1. Nagy G, Koncz A, Perl A. T and B cell abnormalities in systemic lupus erythematosus. Rev Immunolo 2005; 25:123-140.

2. Denny MF, Chandaroy P, Killen PD, Caricchio R, Lewis EE, Richardson BC, et al. Accelerated macrophage apoptosis induces autoantibody formation and organ damage in systemic lupus erythematosus. J Immunol 2006; 176:2095–2104.

3. Cline AM, Radic MZ. Apoptosis, subcellular particles, and autoimmunity. Clin Immunol 2004; 112:175–182.

4. Lemire JM, Ince A, Takashima M. 1,25-Dihydroxy-vitamin D3 attenuates the expression of experimental murine lupus of MRL/l mice. Autoimmunity 1992; 12:143-148.

5. Wahono CS, Rumini H, Soelistyoningsih D, Hakim R, Handono K, Endharti AT, Kalim H, Widjajanto E. Effects of 1,25(OH)2D3 in immune response regulation of systemic lupus erithematosus (SLE) patient with hypovitamin D. Int J Clin Exp Med 2014; 7:22-31.

6. Marques CD, Dantas AT, Fragoso TS, Duarte AL. The importance of vitamin D levels in autoimmune diseases. Bras J Rheumatol 2010; 50:67-80.

7. Vaisberg MW, Kaneno R, Franco MF, Mendes NF: Influence of cholecalciferol (vitamin D3) on the course of experimental systemic lupus erythematosus in F1 (NZBxW) mice. J Clin Lab Anal 2000; 14: 91-96.

8. Lavi Arab F, Rastin M, Faraji F, Zamani Taghizade Rabe S, Tabasi N, Khazaee M, Haghmorad D, Mahmoudi M. Assessment of 1,25-dihydroxyvitamin D3 effects on Treg cells in a mouse model of systemic lupus erythematosus. Immunophar Immunotox 2015; 37:12-18.

9. Amital H, Szekanecz Z, Szücs G, Danko K, Nagy E, Csépany T, et al. Serum concentrations of 25-OH vitamin D in patients with systemic lupus erythematosus (SLE) are inversely related to disease activity: is it time to routinely supplement patients with SLE with vitamin D? Ann Rheum Dis 2010; 69:1155-1157.

10. Contorna MT, Zhu y, Froicu M, Wittke a. Vitamin D status, 1,25-dihydroxyvitamin D3, and the immune system. Am J Clin Nutr 2004; 80:1717S-1720S.

11. Adorini L, Penna G. Control of autoimmune diseases by the vitamin D endocrine system. Nat. Clin. Pract. Rheumatol 2008; 4:404-412.

12. Elen G, Gysemans C, Verlinden L, Vanoirbeek E, De Clercq P, Va Haver D, Mathieu C, Bouillon R, Verstuyf A. Mechanism and potential of the growth inhibitory actions of vitamin D and analogs. Curr Med Chem 2007; 14:1893-1910.

13. Dusso AS, Thadhani R, Slatopolosky e. Vitamin D receptor and analogs. Semin Neph 2004; 24:10-16.

14. Flanagan L, Packman K, Juba B, O Neill S, Tenniswood M, Welsh J. Efficacy of vitamin D compounds to modulate estrogen receptor negative breast cancer growth and invasion. J Estroid Biochem Mol Biol 2003; 84:181-192.

15. Mantel DJ, Owens PE, Bundred NJ, Mawer EB, Canfield AE. 1a, 25-dihydroxy vitamin D3 inhibits angiogenesis in vitro and in vivo. Circ Res 2000; 87:214-220.

16. Verlinden L, Verstuy A, Van Camp M, Marcelis S, Sabbe K, Zhao XY, De Clercq D, Vanderwalle M, Bouillon R. Two novel 14-epi-analogues of 1,25-dihydroxy vitamin D3 inhibit the growth of human
breast cancer cells in vitro and in vivo. Cancer Res 2000; 60: 2673-2679.

17. Muszkat P, Camargo MBR, Griz LHM, Lazaretti-Castro M. “Evidence-based non-skeletal actions of vitamin D,” Arquivos Brasileiros de Endocrinologia eMetabologia 2010; 54:110-117.

18. Zhang H, Shih DQ, Zhang X. Mechanisms under-lying effects of 1,25-Dihydroxy vitamin D3 on the Th17 cells. Eur J Microbiol. Immunol 2013; 3:237-240.

19. Schwalfenberg GK. A review of the critical role of vitamin D in the functioning of the immune system and the clinical implications of vitamin D deficiency. Mol Nutr Food Res 2011; 55:96-108.

20. Navratil JS, Ahearn JM: Apoptosis, clearance mechanisms, and the development of systemic lupus erythematosus. Curr Rheumatol Rep 2001; 3:191-198.

21. Cohen PL. Apoptotic cell death and lupus. Springer Semin Immunopathol 2006; 28: 145-152.

22. Ohnishi T, Takahashi A, Ohnishi K. Studies about space radiation promote new fields in radiation biology. J Radiat Res (Tokyo) 2002; 43 S7-12.

23. Chen S, Sims GP, Chen XX, et al. Modulatory effects of 1,25-dihydroxyvitamin D3 on human B cell differentiation. J Immunol 2007; 179: 1634-1647.

24. Dhir V, Singh AP, Aggarwal A, Naik S, Misra R. Increased T-lymphocyte apoptosis in lupus correlates with disease activity and may be responsible for reduced T-cell frequency: a cross-sectional and longitudinal study. Lupus 2009; 18: 785-791.

25. Okamoto A, Fujio K, Okamura T, Yamamoto K. Regulatory T-cell-associated cytokines in systemic lupus erythematosus. J Biomed Biotechnol 2011:463412.