Anti-inflammatory effects of 1,25-dihydroxyvitamin D3 in monocytes cultured in serum from patients with type 2 diabetes mellitus and diabetic nephropathy with uremia via Toll-like receptor 4 and nuclear factor-κB p65

Yang,Mengxue; Xu,Jie; Yu,Jie; Yang,Bo; Gan,Hua; Li,Sicheng; Li,Xianwen

doi:10.3892/mmr.2015.4482

Anti-inflammatory effects of 1,25-dihydroxyvitamin D3 in monocytes cultured in serum from patients with type 2 diabetes mellitus and diabetic nephropathy with uremia via Toll-like receptor 4 and nuclear factor-κB p65

Authors:
- Mengxue Yang
- Jie Xu
- Jie Yu
- Bo Yang
- Hua Gan
- Sicheng Li
- Xianwen Li
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Affiliations: Department of Endocrinology, The First Affiliated Hospital of Zunyi Medical College, Zunyi, Guizhou 563003, P.R. China, School of Public Health, Zunyi Medical College, Zunyi, Guizhou 563003, P.R. China, Department of Nephrology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: October 23, 2015 https://doi.org/10.3892/mmr.2015.4482
Pages: 8215-8222

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Abstract

Type 2 diabetes mellitus (T2DM) and diabetic nephropathy (DN) seriously affect the quality of human life. 1,25-dihydroxyvitamin D3 (VD3) is known to exert anti‑inflammatory and immunomodulatory effects. The present study investigated the effects of VD3 on Toll‑like receptor 4 (TLR4) and nuclear factor (NF)‑κB p65 expression in monocytes from patients with T2DM and investigated the underlying mechanisms. Serum from subjects of the control, T2DM and DN with uremia groups was isolated, and THP‑1 monocytes were cultured in these sera with or without VD3. After treatment with lipopolysaccharide (LPS) and interleukin (IL)‑15, monocytes and culture supernatants were collected. The expression of TLR4, TLR9 and IL‑15 mRNA was detected using reverse‑transcription polymerase chain reaction analysis. Furthermore, the protein expression of TLR4, NF‑κB p65 and inhibitor of NF‑κB (IκB) was determined using western blot analysis. The levels of IL‑6 and monocyte chemotactic protein in culture supernatants were detected using ELISAs. The impact of LPS, IL‑15 and VD3 on the TLR/NF‑κB signaling pathway in the T2DM and DN uremia groups was also investigated. In the T2DM and DN uremia groups, LPS and IL‑15 downregulated the expression of IκB and upregulated levels of proteins, including TLR4, NF‑κB p65, IL‑6 and monocyte chemoattractant protein 1, as well as TLR4 and IL‑15 mRNA. There was no significant difference in TLR9 mRNA protein expression among the three groups. VD3 partially blocked the above effects; However, pre‑treatment with VD3 had no significant effect on TLR4 mRNA expression, protein expression of NF-κB p65 and IκB, or the levels of associated inflammatory cytokines. In conclusion, the present study indicated that anti‑inflammatory effects of VD3 in inflammatory immune responses in T2DM and DN uremia are associated with the TLR4/NF-κB p65 signaling pathway.

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1	Pickup JC: Inflammation and activated innate immunity in the pathogenesis of type 2 diabetes. Diabetes Care. 27:813–823. 2004. View Article : Google Scholar : PubMed/NCBI
2	Baeke F, Korf H, Overbergh L, van Etten E, Verstuyf A, Gysemans C and Mathieu C: Human T lymphocytes are direct targets of 1,25-dihydroxy1,25(OH)2D3 in the immune system. J Steroid Biochem Mol Biol. 121:221–227. 2010. View Article : Google Scholar : PubMed/NCBI
3	Thacher TD and Clarke BL: Vitamin D insufficiency. Mayo Clin Proc. 86:50–60. 2011. View Article : Google Scholar : PubMed/NCBI
4	Giulietti A, van Etten E, Overbergh L, Stoffels K, Bouillon R and Mathieu C: Monocytes from type 2 diabetic patients have a pro-inflammatory profile 1, 25-dihydroxyvitamin D3 works as anti-inflammatory. Diabetes Res Clin Pract. 77:47–57. 2007. View Article : Google Scholar
5	Du T, Zhou ZG, You S, Huang G, Lin J, Yang L, Li X, Zhou WD and Chao C: Modulation of monocyte hyperresponsiveness to TLR ligands by 1,25-dihydroxy-vitamin D3 from LADA and T2DM. Diabetes Res Clin Pract. 83:208–214. 2009. View Article : Google Scholar
6	Liu PT, Steffen S, Li H, Wenzel L, Tan BH, Krutzik SR, Ochoa MT, Schauber J, Wu K, Meinken C, et al: Toll-like receptor triggering of a vitamin D mediated human antimicrobial response. Science. 311:1770–1773. 2006. View Article : Google Scholar : PubMed/NCBI
7	Nimitphong H, Holick MF, Fried SK and Lee MJ: 25-hydroxyvitamin D₃ and 1,25-dihydroxyvitamin D₃ promote the differentiation of human subcutaneous preadipocytes. PLoS One. 7:e521712012. View Article : Google Scholar
8	Kim F, Pham M, Luttrell I, Bannerman DD, Tupper J, Thaler J, Hawn TR, Raines EW and Schwartz MW: Toll-like receptor 4 mediates vascular inflammation and insulin resistance in diet-induced obesity. Circ Res. 100:1589–1596. 2007. View Article : Google Scholar : PubMed/NCBI
9	Devaraj S, Dasu MR, Rockwood J, Winter W, Griffen SC and Jialal I: Increased Toll-like receptor (TLR) 2 and TLR4 expression in monocytes from patients with type 1 diabetes: further evidence of a proinflammatory state. J Clin Endocrinol Metab. 93:578–583. 2008. View Article : Google Scholar
10	Dasu MR, Devaraj S, Park S and Jialal I: Increased Toll-like receptor (TLR) activation and TLR ligands in recently diagnosed Type 2 diabetic subjects. Diabetes Care. 33:861–868. 2010. View Article : Google Scholar : PubMed/NCBI
11	Verma R, Jung JH and Kim JY: 1,25-Dihydroxyvitamin D3 upregulates TLR10 while downregulating TLR2, 4 and 5 in human monocyte THP-1. J Steroid Biochem Mol Biol. 141:1–6. 2014. View Article : Google Scholar : PubMed/NCBI
12	Sadeghi K, Wessner B, Laggner U, Ploder M, Tamandl D, Friedl J, Zügel U, Steinmeyer A, Pollak A, Roth E, et al: Vitamin D3 downregulates monocyte TLR expression and triggers hyporesponsiveness to pathogen-associated molecular patterns. Eur J Immunol. 36:361–370. 2006. View Article : Google Scholar : PubMed/NCBI
13	Gu HR, Luan B, Qiao JY, Wang YZ and Li Q: Effect of 1,25-(OH)2D3 on expression of HMGB1 and TLR4 in the lungs of asthmatic mice. Zhongguo. Dang Dai Er Ke Za Zhi. 16:301–305. 2014.In Chinese.
14	Li F, Liu P, Zhang X, Zhang Q, Tang S, Zhu M and Qiu M: 1,25(OH)2D3-mediated amelioration of aortic injury in strep-tozotocin-induced diabetic rats. Inflammation. 36:1334–1343. 2013. View Article : Google Scholar : PubMed/NCBI
15	Yang M, Gan H, Shen Q, Tang W, Du X and Chen D: Proinflammatory CD14+CD16+monocytes are associated with microinflammation in patients with type 2 diabetes mellitus and diabetic nephropathy uremia. Inflammation. 35:388–396. 2012. View Article : Google Scholar
16	Yang M, Shen Z, Chen D, et al: Effects of 1,25-(OH)2D3 on the expressions of vitamin D receptor, STAT5 and cytoskeletal rearrangement in human monocytes incubated with sera from type 2 diabetes patients and diabetic nephropathy patients with uremia. Inflamm Res. 61:511–520. 2012. View Article : Google Scholar : PubMed/NCBI
17	Yang M, Gan H, Shen Q, et al: Proportion of CD14+CD16+ monocytes in peripheral blood from patients with type 2 diabetic mellitus and effect of LPS and IL-15 on expression of STAT5 in monocytes. Chinese Pathophysiology. 1:136–141. 2012.
18	Alleva DG, Kaser SB, Monroy MA, Fenton MJ and Beller DI: IL-15 functions as a potent autocrine regulator of macrophage proinflammatory cytokine production: Evidence for differential receptor subunit utilization associated with stimulation or inhibition. J Immunol. 159:2941–2951. 1997.PubMed/NCBI
19	Yao L, Xiao Y, Liu SP, Xu AM and Zhou ZG: Serum of obesity induce the activation of TLR4/NF-κB signaling pathway on THP-1 cell line. Zhonghua Yi Xue Za Zhi. 90:3119–3123. 2010.In Chinese.
20	Ge JB and Xu YJ: Internal Medicine. Edition 8. People Health Publishing House; Beijing: pp. 7412013
21	Pramanik R, Asplin JR, Lindeman C, Favus MJ, BAi S and Coe FL: Lipopolysaccharide negatively modulates vitamin D action by downregulating expression of vitamin D-induced VDR in human monocytic THP-1 cells. Cell Immunol. 232:137–143. 2004. View Article : Google Scholar
22	Sadeghi K, Wessner B, Laggner U, Ploder M, Tamandl D, Friedl J, Zügel U, Steinmeyer A, Pollak A, Roth E, et al: Vitamin D3 downregulates monocyte TLR expression and triggers hypo-responsiveness to pathogen-associated molecular patterns. Eur J Immunol. 36:361–370. 2006. View Article : Google Scholar : PubMed/NCBI
23	Navarro-González JF and Mora-Fernández C: The role of inflammatory cytokines in diabetic nephropathy. J Am Soc Nephrol. 19:433–442. 2008. View Article : Google Scholar : PubMed/NCBI
24	Ortiz-Muñoz G, Lopez-Parra V, Lopez-Franco O, Fernandez-Vizarra P, Mallavia B, Flores C, Sanz A, Blanco J, Mezzano S, Ortiz A, et al: Suppressors of cytokine signaling abrogate diabetic nephropathy. J Am Soc Nephrol. 21:763–772. 2010. View Article : Google Scholar : PubMed/NCBI
25	O'Connor JC, Satpathy A, Hartman ME, Horvath EM, Kelley KW, Dantzer R, Johnson RW and Freund GG: IL-1beta-mediated innate immunity is amplified in the db/db mouse model of type 2 diabetes. J Immunol. 174:4991–4997. 2005. View Article : Google Scholar : PubMed/NCBI
26	Mora C and Navarro JF: Inflammation and diabetic nephropathy. Curr Diab Rep. 6:463–468. 2006. View Article : Google Scholar : PubMed/NCBI
27	Navarro JF and Mora C: Diabetes, inflammation, proinflammatory cytokines and diabetic nephropathy. Scientific World Journal. 6:908–917. 2006. View Article : Google Scholar
28	Tuttle KR: Linking metabolism and immunology: Diabetic nephropathyis an inflammatory disease. J Am Soc Nephrol. 16:1537–1538. 2005. View Article : Google Scholar : PubMed/NCBI
29	Ruster C and Wolf G: The role of chemokines and chemokine receptors in diabetic nephropathy. Front Biosci. 13:944–955. 2008. View Article : Google Scholar
30	Stubbs JR, Idiculla A, Slusser J, Menard R and Quarles LD: Cholecalciferol supplementation alters calcitriol-responsive monocyte proteins and decreases inflammatory cytokines in ESRD. J Am Soc Nephrol. 21:353–361. 2010. View Article : Google Scholar :
31	Akira S and Takeda K: Toll-like receptor signaling. Nat Rev Immunol. 7:499–511. 2004. View Article : Google Scholar
32	Cao XT: The Progress of Immunology. People's Medical Publishing House; Beijing: pp. 1432009
33	Li ML, Gan H and Qiao L: The expression of TLR4 on peripheral blood monocytes from uremic patients with diabetic nephropathy and its relation with plasma MCP-1 concentration. Chinese Journal of Immunology. 9:848–850. 2009.