Conventional but not High-Purity Dialysis Fluid Induces Apoptosis and Insulin Resistance in Adipocytes in Vitro

Abstract

Purpose:

Previous study has shown an independent association of low body fat percentage and fat loss over time with higher mortality in hemodialysis (HD) patients. High-purity dialysis fluid (HPDF) used in place of conventional dialysis fluid (CDF) may decrease inflammation and improve nutritional status. Its effect on adipocytes and therefore fat storage is unclear. The purpose of this study was to assess the effects of these dialysates on apoptosis and insulin resistance in cultured adipocytes to determine a basis for the superiority of HPDF.

Methods:

Flow cytometry and fluorescence staining were used to evaluate apoptosis in adipocytes. Insulin receptor substrate-1 (IRS-1) and −2 and serine phosphorylation of IRS-1 levels were assayed by Western blot. Tumor necrosis factor α (TNFα) levels were measured by enzyme-linked immunosorbent assay (ELISA).

Results:

Although CDF contained more bacteria (66 ± 6 CFU/mL) than HPDF (7 ± 3 CFU/mL) and higher levels of bacterial DNA, endotoxin levels were similar. Combined with uremic serum (containing high TNFα levels), CDF (vs. HPDF) induced more apoptosis in adipocytes. Exposure to CDF also decreased expression of IRS-1 protein, increased expression of IRS-2 protein and insulin-stimulated serine phosphorylation of IRS-1, and decreased glucose consumption. Additionally, the culture supernatant from adipocytes exposed to CDF plus uremic serum for 48 hours contained significantly higher levels of TNFα (15.17 ± 2.89 vs. 8.12 ± 0.54 pg/mL [HPD plus uremic serum], p<0.0001).

Conclusion:

The advantage of HPDF over CDF may be due to lower levels of dialysis fluid impurities.

Keywords

Apoptosis Hemodialysis High-purity dialysis fluid Insulin resistance 3T3-L1 adipocyte

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References

Abbott

K.C.

, Glanton

C.W.

, Trespalacios

F.C.

Body mass index, dialysis modality, and survival: analysis of the United States Renal Data System Dialysis Morbidity and Mortality Wave II Study.

Kidney Int. 2004; 65(2): 597–605.

Plantinga

L.C.

, Fink

N.E.

, Levin

N.W.

Early, intermediate, and long-term risk factors for mortality in incident dialysis patients: the Choices for Healthy Outcomes in Caring for ESRD (CHOICE) Study.

Am J Kidney Dis. 2007; 49(6): 831–840.

Lonnemann

. When good water goes bad: how it happens, clinical consequences and possible solutions. Blood Purif. 2004; 22(1): 124–129.

Schindler

. Inflammation and dialysate quality. Hemodial Int. 2006; 10(Suppl 1): S56–S59.

Schiffl

, Lang

S.M.

, Stratakis

, Fischer

. Effects of ultrapure dialysis fluid on nutritional status and inflammatory parameters. Nephrol Dial Transplant. 2001; 16(9): 1863–1869.

Zhang

D.H.

, Yu

X.Q.

, Zheng

Z.H.

Effects of high purity dialysis concentrate on the levels of serum IL-6, TNFα and albumin in hemodialysis patients.

Chin J Nephrol. 2005; 21: 76–80.

Guo

L.L.

, Pan

, Zhu

X.J.

Conventional, but not high-purity, dialysate-induced monocyte apoptosis is mediated by activation of PKC-{delta} and inflammatory factors release.

Nephrol Dial Transplant. 2011; 26(5): 1516–1522.

Kalantar-Zadeh

, Kuwae

, Wu

D.Y.

Associations of body fat and its changes over time with quality of life and prospective mortality in hemodialysis patients.

Am J Clin Nutr. 2006; 83(2): 202–210.

Schindler

, Beck

, Deppisch

Short bacterial DNA fragments: detection in dialysate and induction of cytokines.

J Am Soc Nephrol. 2004; 15(12): 3207–3214.

10.

Bossola

, Sanguinetti

, Scribano

Circulating bacterial-derived DNA fragments and markers of inflammation in chronic hemodialysis patients.

Clin J Am Soc Nephrol. 2009; 4(2): 379–385.

11.

Lin

, Page

K.A.

, Della-Fera

M.A.

, Baile

C.A.

. Evaluation of adipocyte apoptosis by laser scanning cytometry. Int J Obes Relat Metab Disord. 2004; 28(12): 1535–1540.

12.

Yin

, Hu

, Chen

Effects of berberine on glucose metabolism in vitro.

Metabolism. 2002; 51(11): 1439–1443.

13.

Galli

, Ghibelli

, Buoncristiani

Mononuclear leukocyte apoptosis in haemodialysis patients: the role of cell thiols and vitamin E.

Nephrol Dial Transplant. 2003; 18(8): 1592–1600.

14.

Prins

J.B.

, Niesler

C.U.

, Winterford

C.M.

Tumor necrosis factor-alpha induces apoptosis of human adipose cells.

Diabetes. 1997; 46(12): 1939–1944.

15.

Matsumoto

, Sera

, Abe

, Ueki

, Tominaga

, Miyake

. Inflammation and insulin resistance are independently related to all-cause of death and cardiovascular events in Japanese patients with type 2 diabetes mellitus. Atherosclerosis. 2003; 169(2): 317–321.

16.

Rasić-Milutinović

, Perunicić-Peković

, Ristić-Medić

, Popović

, Glibetić

, Djurić

D.M.

. Insulin resistance and chronic inflammation are associated with muscle wasting in end-stage renal disease patients on hemodialysis. Gen Physiol Biophys. 2009; 28: 184–189.

17.

Tanti

J.F.

, Jager

. Cellular mechanisms of insulin resistance: role of stress-regulated serine kinases and insulin receptor substrates (IRS) serine phosphorylation. Curr Opin Pharmacol. 2009; 9(6): 753–762.

18.

Morino

, Neschen

, Bilz

Muscle-specific IRS-1 Ser->Ala transgenic mice are protected from fat-induced insulin resistance in skeletal muscle.

Diabetes. 2008; 57(10): 2644–2651.

19.

Sasaki-Suzuki

, Arai

, Ogata

Growth hormone inhibition of glucose uptake in adipocytes occurs without affecting GLUT4 translocation through an insulin receptor substrate-2-phosphatidylinositol 3-kinase-dependent pathway.

J Biol Chem. 2008; 284(10): 6061–6070.

20.

Sakoda

, Ogihara

, Anai

Dexamethasone-induced insulin resistance in 3T3-L1 adipocytes is due to inhibition of glucose transport rather than insulin signal transduction.

Diabetes. 2000; 49(10): 1700–1708.

21.

de Alvaro

, Teruel

, Hernandez

, Lorenzo

. Tumor necrosis factor alpha produces insulin resistance in skeletal muscle by activation of inhibitor kappa B kinase in a p38 MAPK-dependent manner. J Biol Chem. 2004; 279(17): 17070–17078.

22.

Gao

, Zuberi

, Quon

M.J.

, Dong

, Ye

. Aspirin inhibits serine phosphorylation of insulin receptor substrate 1 in tumor necrosis factor-treated cells through targeting multiple serine kinases. J Biol Chem. 2003; 278(27): 24944–24950.

23.

Wieczorowska-Tobis

, Polubinska

, Breborowicz

, Oreopoulos

D.G.

. A comparison of the biocompatibility of phosphate-buffered saline and dianeal 3.86% in the rat model of peritoneal dialysis. Adv Perit Dial. 2001; 17: 42–46.

24.

Wieczorowska-Tobis

, Styszynski

, Polubinska

, Radkowski

, Breborowicz

, Oreopoulos

D.G.

. Hypertonicity of dialysis fluid suppresses intraperitoneal inflammation. Adv Perit Dial. 2000; 16: 262–266.

25.

Wieczorowska-Tobis

, Styszynski

, Breborowicz

, Oreopoulos

D.G.

. Comparison of the biocompatibility of phosphate-buffered saline alone, phosphate-buffered saline supplemented with glucose, and dianeal 3.86%. Perit Dial Int. 2001; 21(Suppl 3): S362–S364.