Relationship between glycemic control and histochemical myeloperoxidase activity in neutrophils in patients with type 2 diabetes

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1 DOI /s Dibetology & Metbolic Syndrome RESEARCH Open Access Reltionship between glycemic control nd histochemicl myeloperoxidse ctivity in neutrophils in ptients with type 2 dibetes Mustf Unubol 1*, Irfn Yvsoglu 2, Firuzn Kcr 3, Engin Guney 1, Imrn Kurt Omurlu 4, Mevlut Ture 4, Gurhn Kdikoylu 2 nd Zhit Bolmn 2 Abstrct Bckground: Myeloperoxidse (MPO) is lysosoml hemoprotein found in the zurophilic grnules in neutrophils. Myeloperoxidse plys n importnt role in oxygen-dependent killing of bcteri, fungi, virus nd mlignnt cells. Dibetes mellitus (DM) is listed mong conditions tht my led to secondry MPO deficiency in neutrophils but inconsistent results concerning MPO ctivity in dibetic ptients hve been reported in the literture. In this study, we imed to evlute the reltionship between glycemic control in ptients with type 2 DM nd MPO ctivity in neutrophils from histochemicl perspective. Methods: The study included 40 ptients with type 2 DM with poor glycemic control, 30 ptients with type 2 DM with good glycemic control nd 31 helthy controls. Peripherl blood smers were nlyzed for ech ptient included in the study. Myeloperoxidse dye ws used for stining. Myeloperoxidse rtios in neutrophil were evluted for proportions of stining with MPO in 100 neutrophils in ech smer. SPSS 16.0 version ws used for sttisticl nlyses. Results: Myeloperoxidse rtios in neutrophils were 70 ( ) in type 2 DM ptients with poor glycemic control compred to 80 ( ) in those with good glycemic control nd 88 (78 92) in helthy controls. The DM group with poor glycemic control ws sttisticlly significntly different from the other groups (p < 0.001). Conclusions: Poor glycemic control in dibetic ptients results in decresed MPO ctivity in neutrophils histochemiclly. Keywords: Dibetes mellitus, Myeloperoxidse ctivity, Glycemic control, Dibetic complictions Bckground Severl microbicidl dysfunctions of the neutrophils nd monocytes hve been defined s fctor tht contributes to complictions nd to morbidity nd mortlity in dibetic ptients [1, 2]. Deficiencies in number of functions hve been demonstrted in the neutrophils of dibetic ptients [3 5]. Oxidtive stress nd rective oxygen species re known to hve importnt roles in the etiology of dibetes, in development of its complictions nd in disese progression [6]. Myeloperoxidse (MPO) is *Correspondence: drmunubol@yhoo.com.tr 1 Deprtment of Endocrinology, Fculty of Medicine, Adnn Menderes University, Aydın, Turkey Full list of uthor informtion is vilble t the end of the rticle lysosoml hemoprotein found in the zurophilic grnules in neutrophils [7]. It is potent bctericidl enzyme tht produces rective oxygen species. Myeloperoxidse plys n importnt role in oxygen-dependent killing of bcteri, fungi, virus nd mlignnt cells. Myeloperoxidse-medited dmge is not limited to intrphgosoml microbes. It is involved in the pthogenesis of severl inflmmtory conditions, therosclerosis, demyelinting diseses of the centrl nervous system nd some tumors [8 10]. Dibetes mellitus (DM) is listed mong conditions tht my led to secondry MPO deficiency in neutrophils [9] but inconsistent results concerning MPO ctivity in dibetic ptients hve been reported in the literture [11 13]. Our literture scn demonstrted tht the effect of 2015 Unubol et l. This rticle is distributed under the terms of the Cretive Commons Attribution 4.0 Interntionl License ( which permits unrestricted use, distribution, nd reproduction in ny medium, provided you give pproprite credit to the originl uthor(s) nd the source, provide link to the Cretive Commons license, nd indicte if chnges were mde. The Cretive Commons Public Domin Dediction wiver ( publicdomin/zero/1.0/) pplies to the dt mde vilble in this rticle, unless otherwise stted.

2 Pge 2 of 7 glycemic control in dibetic ptients on MPO ctivity in neutrophils hs not been investigted from histochemicl perspective previously. In this study, we imed to evlute the reltionship between glycemic control in ptients with type 2 DM nd MPO ctivity in neutrophils from histochemicl perspective. Methods Study design nd protocol This study ws designed s n observtionl, cross-sectionl nd cse controlled study. Ptients with type 2 DM who presented to outptient clinics of the Adnn Menderes University, Medicl Fculty, Deprtment of Endocrinology nd Metbolic Disese between Jnury nd Mrch 2012 were studied. The ptients with type 2 DM pplying to the follow-up clinic, nd lcking the exclusion criteri, greeing to prticipte in the study were enrolled consecutively from Jnury The study included 40 ptients with type 2 DM with poor glycemic control (HbA1c > 7), 30 ptients with type 2 DM with good glycemic control (HbA1c 7) [14] nd 31 helthy controls. Ptients with ctive infection, known mlignncy, hemtologic mlignncy, iron deficiency, thrombotic conditions, renl trnsplnttion, ptients currently treted with cytotoxic gents, fenofibrte, nd nti-inflmmtory drugs, nd pregnnt women were excluded from the study. Three peripherl blood smers (in fsting period) were nlyzed for ech ptient included in the study. Myeloperoxidse dye ws used for stining. Myeloperoxidse rtios in neutrophils (Fig. 1) were evluted by one hemtologist nd one hemtopthologist for Fig. 1 Microscopic imge of the neutrophil with positive MPO stining in peripherl blood proportions of stining with MPO in 100 neutrophils in ech smer, nd the men vlue of the redings were reported [15]. Sttisticl nlysis SPSS 16.0 version ws used for sttisticl nlyses. Fit of the quntittive dt to norml distribution ws studied using the Shpiro Wilk test. For vribles tht were fit for norml distribution, one-wy vrince nlysis (ANOVA) ws used for sttisticl comprisons nd descriptive sttistics were expressed s men ± stndrd devition. For vribles unfit for norml distribution, Kruskl Wllis ws used for sttisticl comprisons nd descriptive sttistics were expressed s medin (25 75 ). The level of reltionship between vribles ws studied with Spermn s Rho correltion coefficient. Chi squre test nd Fisher test were used in nlyzing ctegoricl vribles nd descriptive sttistics were expressed s frequency (%). In our study, we used clssifiction nd regression tree (C&RT) method in order to choose the best predictor for MPO. p vlues < 0.05 were considered sttisticlly significnt. Approvl of the ethics bord of the Medicl Fculty of Adnn Menderes University ws received. Results Type 2 DM ptients with poor glycemic control hd men ge of 56.4 ± 9.99 yers nd type 2 DM ptients with good glycemic control hd men ge of 57.3 ± yers nd helthy controls hd men ge of ± yers, with no significnt difference mong the groups (p > 0.05). Type 2 DM ptients with poor glycemic control hd disese durtion of 12.3 ± 3.4 yers nd were not significntly different from type 2 DM ptients with good glycemic control (p > 0.05). Myeloperoxidse rtio in neutrophils ws 70 ( ) in type 2 DM ptients with poor glycemic control compred to 80 ( ) in those with good glycemic control nd 88 (78 92) in helthy controls. The DM group with poor glycemic control ws sttisticlly significntly different from the other groups (p < 0.001). Good nd poor controlled dibetes groups were similr in terms of the drugs being used. No correltion ws found between the MPO ctivity nd the drugs used to tret dibetes (p > 0.05). In ptient groups with dibetes, sttisticlly significnt differences in terms of MPO ctivity ws not detected mong the ptients using nd not using sitgliptin, insulin detemir, insulin glrgine, insulin sprt, insulin lispro, insulin glulisine, metformin, gliclzide, repglinide, nteglinide, pioglitzone. Sttisticl results of quntittive nd qulittive dt re provided in Tbles 1, 2, 3 nd 4; Figs. 2 nd 3 demonstrtes correltion nlysis between MPO nd HbA1c.

3 Pge 3 of 7 Tble 1 Comprison of lbortory findings between the three groups DM with poor glycemic control DM with good glycemic control Controls p MPO (%) 70 ( ) 80 ( ) 88 (78 92) <0.001 b Hb (g/dl) ± ± ± 1.47 >0.05 c Leukocyte (/mm 3 ) 7961 ± ± ± d c Pltelet (/mm 3 ) ± ± ± >0.05 c HBA1C (%) ( ) 6.5 ( ) e 5.4 (5 5.5) <0.001 b LDL (mg/dl) ± ± ± >0.05 c HDL (mg/dl) ± f ± ± c Triglyceride (mg/dl) ( ) ( ) 120 (87 190) >0.05 b Cretinine (mg/dl) 0.77 ( ) 0.72 ( ) 0.73 ( ) b Fsting plsm glucose (mg/dl) ( ) 122 ( ) e 90 (85 93) <0.001 b Postprndil plsm glucose (mg/dl) 298 ( ) ( ) e 104 (95 120) <0.001 b Sedimenttion (mm/h) ± g ± ± c C-rective protein (g/dl) 3.41 ( ) 1.35 ( ) 1.38 ( ) b Age 56.4 ± ± ± >0.05 c BMI (kg/m 2 ) ( ) ( ) 25 ( ) d b MPV (fl) 9.4 ( ) 8.4 ( ) 8.9 ( ) >0.05 b Hemtocrite ( ) 40.6 ( ) 36.7 ( ) h b 24-h urine microlbuminuri (mg/dy) 38 ( ) 17 ( ) e 12 (8 15) <0.001 b DM group with poor control is different from other groups b It ws used the Kruskl Wllis test for sttisticl nlysis c It ws used the ANOVA test for sttisticl nlysis d Control group is different from other groups e All groups re different from ech other f DM group with poor control nd DM group with good control re different from ech other g DM group with poor control nd control group re different from ech other h Control group nd DM group with good control re different from ech other We pllied the C&RT model in order to choose the best predictor for MPO development. In the model, ptients were first divided into two brnches ccording to their type 2 DM vlues (ptients with poor glycemic control nd good glycemic control nd helthy controls) for the prediction of MPO. According to this model, type 2 DM ptients with poor glycemic control hd lower MPO ctivity in neutrophils (66.6 ± 21.6 versus 81.9 ± 14.0). MPO impct on the dignosis of Type 2 DM ws 100 % nd the effects of drugs used to tret DM nd HT ws 63 %. Discussion We determined histochemiclly tht type 2 DM ptients with poor glycemic control hd lower MPO ctivity in neutrophils compred to both helthy controls nd type 2 DM ptients with good glycemic control. Type 2 DM ptients with good glycemic control nd helthy controls did not differ significntly with respect to MPO ctivity in neutrophils. Myeloperoxidse deficiency in neutrophils cn be cquired or hereditry. Acquired MPO deficiency is rrer [10]. Hemtologic mlignncies, common cncers, cytotoxic gents, some ntiinflmmtory drugs, iron deficiency, led intoxiction, thrombotic conditions, renl trnsplnttion, serious infections, neuronl lipofuscinosis, pregnncy nd DM hve been shown mong the cuses of cquired MPO deficiency [8, 10]. Nit et l. showed tht fenofibrte reduced plsm MPO concentrtions [16]. A study tht compred MPO rtios in peripherl neutrophils in dibetic ptients identified no significnt difference in MPO rtios of dibetic ptients with nd without infection [11]. In the study by Sto et l. [12], decresed MPO ctivity ws shown in leukocytes of dibetic ptients. Likewise, study by Uchimur et l. [17] demonstrted mrkedly decresed MPO ctivity in leukocytes of ptients with non-insulin dependent dibetes. On the other hnd, Gorudko et l. [13] found incresed MPO ctivity in plsm of type 2 DM ptients with no crdiovsculr disese. In the study by Moldovenu et l. [18], ptients with DM lso hd higher MPO ctivity compred to the control group. In nother study, serum MPO levels were found to be higher in overweight individuls who hd first-degree reltives with type 2 DM compred to the control group [19]. An niml study demonstrted tht MPO expression ws higher in

4 Pge 4 of 7 Tble 2 Comprison of neuropthy presence nd lbortory dt in ptients with type 2 DM With neuropthy (n = 27) Without neuropthy (n = 43) Controls (n = 31) p MPO (%) 75 (50 86) 80 (70 90) 87 ( ) b HBA1C (%) 9 ( ) c 6.75 ( ) 5.4 ( ) <0.001 b Sedimenttion (mm/h) ± d ± ± e C-rective protein (g/dl) 4.1 ( ) d 1.95 ( ) 1.37 ( ) b b The group with neuropthy nd the control group re different from ech other It ws used the Kruskl Wllis test for sttisticl nlysis c All groups re different from ech other d The group with neuropthy is different from other groups e It ws used the ANOVA test for sttisticl nlysis Tble 3 Comprison of nephropthy development nd lbortory dt in ptients with type 2 DM With nephropthy (n =29) Without nephropthy (n = 41) Controls (n =31) p MPO (%) 80 (70 85) 75 (54 88) 88 (78 92) b HBA1C (%) 9 (7.1 12) 6.9 ( ) 5.4 (5 5.5) <0.001 b Sedimenttion (mm/h) 38 ± ± ± c d C-rective protein (g/dl) 3.44 ( ) 2.41 ( ) 1.38 ( ) e b Control group is different from other groups b It ws used the Kruskl Wllis test for sttisticl nlysis c Control group nd the group without nephropthy re different from ech other d It ws used the ANOVA test for sttisticl nlysis e Control group nd the group with nephropthy re different from ech other Tble 4 Comprison of complictions presence in ptients with type 2 DM Controls n (%) DM with poor control n (%) DM with good control n (%) Exct p Sex Mle 8 (25.8) 21 (52.5) 11 (36.7) >0.05 Femle 23 (74.2) 19 (47.5) 19 (63.3) Neuropthy No 20 (50) 23 (76.7) <0.001 Yes 20 (50) 7 (23.3) CAD No 32 (80) 28 (93.3) <0.001 Yes 8 (20) 2 (6.7) Retinopthy No 31 (77.5) 28 (93.3) <0.001 Yes 9 (22.5) 2 (6.7) PAH No 38 (95) 29 (96.7) <0.001 Yes 2 (5) 1 (3.3) Dibetic foot No 38 (95) 30 (100) >0.05 Yes 2 (5) 0 (0) Fisher test were used in nlyzing ctegoricl vribles nd descriptive sttistics were expressed s frequency (%) It ws used the Chi squre test for sttisticl nlysis dibetic neutrophils but reported decresed MPO ctivity in neutrophils compred to the control group [20]. The reltionship between type 2 dibetes mellitus nd MPO hs been investigted by previous studies, which rrived t contrdicting conclusions [13, 17 19]. There re limited number of studies investigting the reltionship between glycemic response nd MPO. The study by Sto et l. evluting the effect of glycemic response on MPO demonstrted significnt correltion between decresed MPO ctivity in leukocytes nd incresed

5 Pge 5 of 7 Fig. 2 Correltion between HbA1c nd MPO in ptients with DM Fig. 3 Correltion between HbA1c nd MPO in ll groups HbA1c [12]. In our study, we lso found negtive correltion between Hb1C vlues nd MPO levels. Previous studies investigting the reltionship between DM nd MPO used the ELISA method [12, 13, 17 20]. With the literture scn, our study ppers s the first to evlute MPO ctivity in neutrophils in dibetic ptients from histochemicl perspective. Histochemicl evlution of MPO ctivities is cheper nd esier thn serologicl methods. Chronic hyperglycemi increses the relese of rective oxygen species from neutrophils [21]. Neutrophil dysfunctions were shown s one of the cuses of glycemic unresponsiveness in dibetic ptients. Abnormlities in grnulocyte chemotxis, phgocytosis nd microbicidl ctivities were described for ptients with poorlycontrolled dibetes [22]. Impired neutrophil bctericidl function ws strongly ssocited with poor glycemic control nd improved positively with good glycemic control [23]. The chronic hyperglycemi of poorly controlled dibetes cn prime neutrophils nd monocytes [24]. A positive correltion between HbA1c nd white blood cell levels (WBC) in ptients with type 2 DM ws reported in study [25]. Ptients with type 2 DM re in stte of low-degree chronic inflmmtion tht induces hypersecretion of inflmmtory fctors, which results in constntly elevted neutrophilic grnulocyte count [26]. In our study, WBC levels of DM groups with poor glycemic control nd good glycemic control were sttisticlly significntly elevted from the control group (p < 0.001). In study, superoxide dismutse ctivity in the neutrophils of type 2 dibetes ptients ws decresed by 41 % compred to the control group. Glutthione peroxidse (GSHPx) nd glutthione reductse (GR) ctivities of type 2 dibetic ptients were nd %, respectively, compred to controls. No differences were noted in ctlse ctivities. It hs been suggested tht these findings could ccount for some of the mechnisms tht led to incresed sensitivity of type 2 DM ptients to certin infections [27]. Incresed glucose levels led to incresed protein glyction through erly glyction products nd dvnced glyction lte products (AGE) in dibetic ptients. This plys n importnt role in occurrence of complictions [28]. In ddition, AGE is one of the most importnt cuses by which hyperglycemi cuses cellulr nd tissue dmge. Some AGEs occur on criticl protein sites nd my led to enzymtic inctivtion nd loss of physiologicl function [29]. Thus, decresed MPO ctivity in ptients with poorly controlled dibetes my be due to hyperglycemissocited negtive modultion of the enzymtic ctivity [20]. This hypothesis ws described for other enzymes previously. It suggests tht the presence of high plsm glucose concentrtion might cuse ltertions in the moleculr conformtion of the enzyme or its ctlytic site, possibly by glyction of mino cids [30]. There re vrying opinions nd results in the literture regrding the conditions tht low MPO ctivity in neutrophils could led to [31, 32]. Is MPO friend or foe? [31]. It hs been suggested tht chronic inflmmtory process nd serious infections my be more frequent in ptients with totl or subtotl MPO deficiency [32]. Different publictions hve reported tht serious infections s spordic cses developed in less thn 5 % of the individuls with MPO deficiency [8]. While it hs been suggested tht there might be reltionship between MPO

6 Pge 6 of 7 deficiency nd cncer risk [8], different publiction hs reported no increse in the incidence of cncer in individuls with MPO deficiency [31]. It hs been suggested tht MPO deficiency hs crdioprotective effects [32]. Becuse this ws cross-sectionl study, informtion on ptients infections nd mlignncies were derived from ptient reports. The reltionship between MPO deficiency nd infections or mlignncies could not be evluted s conclusive dt could not be obtined. We believe tht evlution by prospective studies on the clinicl conditions tht could be cused by MPO deficiency we observed in ptients with poor glycemic control would be helpful. There re severl studies evluting the reltionship between MPO deficiency nd therosclerosis [33, 34]. In our study, sttisticlly significnt difference ws not noted in the MPO rtio between DM ptients with nd without neuropthy (p > 0.05). Also, no significnt difference ws observed between DM ptients with nd without nephropthy with respect to MPO levels (p > 0.05). In our study, sttisticl evlution could not be performed since the number of ptients with coronry rtery disese nd dibetic rethinopthy in ech group were insufficient. This ws interpreted s finding tht differs from the findings of studies which demonstrted protective effects of deficient levels of MPO in neutrophils from therosclerosis [33, 34]. The role of oxidtive stress in development of dibetic microvsculr complictions is known [35] but poor glycemic control is the primry cuse in development of the microvsculr complictions [36]. The study by Zhng demonstrted tht MPO ws not necessry to induce n experimentl therosclerosis model in MPO-deficiency-induced mice [37]. Our study indictes tht microvsculr complictions in dibetic ptients cnnot be ssocited lone with MPO ctivity. Our study did not yield ny finding which suggests tht decresed MPO ctivity in neutrophils in dibetic ptients prevented microvsculr complictions. In conclusion, poor glycemic control in dibetic ptients results in decresed MPO ctivity in neutrophils histochemiclly. We believe tht vribility in MPO ctivity by glycemic response in dibetic ptients, complictions nd their clinicl relevnce need to be ssessed in prospective studies. Authors contributions MU nd IY conceptulized the ide for the study, collected the dt, performed literture review, nd wrote the mnuscript. MU, IY, EG nd FK prticipted in the design of the study, prticipted in the discussion, nd were involved in drfting the mnuscript. GK, IKO nd MT were involved in performing the sttisticl nlysis. GK, FK, EG nd ZB prticipted in the discussion. All uthors hve red nd pproved the finl mnuscript. Author detils 1 Deprtment of Endocrinology, Fculty of Medicine, Adnn Menderes University, Aydın, Turkey. 2 Deprtment of Hemtology, Fculty of Medicine, Adnn Menderes University, Aydın, Turkey. 3 Deprtment of Pthology, Fculty of Medicine, Adnn Menderes University, Aydın, Turkey. 4 Deprtment of Biosttistics, Fculty of Medicine, Adnn Menderes University, Aydın, Turkey. Competing interests The uthors declre tht they hve no competing interests. Experimentl power In one-wy ANOVA study, smple sizes of 30, 30, nd 30 re obtined from the three groups whose mens re to be compred. The totl smple of 90 subjects chieves 98 % power to detect differences mong the mens versus the lterntive of equl mens using n F test with 0.05 significnce level. Funding This reserch did not receive ny specific grnt from ny funding gency in the public, commercil or not-for-profit sector. Received: 1 September 2015 Accepted: 19 December 2015 References 1. Htnk E, Montegudo PT, Mrrocos MS, Cmp A. Neutrophils nd monocytes s potentilly importnt sources of proinflmmtory cytokines in dibetes. Clin Exp Immunol. 2006;146: Herrer BS, Hsturk H, Kntrci A, Freire MO, Nguyen O, Knsl S, et l. Impct of resolvin E1 on murine neutrophil phgocytosis in type 2 dibetes. Infect Immun. 2015;83: de Vries MA, Alipour A, Klop B, vn de Geijn GJ, Jnssen HW, Njo TL, et l. Glucose-dependent leukocyte ctivtion in ptients with type 2 dibetes mellitus, fmilil combined hyperlipidemi nd helthy controls. Metbolism. 2015;64: Krol E, Agueel R, Bnue S, Smogorzewski M, Kumr D, Mssry SG. Amlodipine reverses the elevtion in [C2 +]i nd the impirment of phgocytosis in PMNLs of NIDDM ptients. Kidney Int. 2003;64: Sefil F, Uluts KT, Dokuyucu R, Sumbul AT, Yengil E, Ygiz AE, et l. Investigtion of neutrophil lymphocyte rtio nd blood glucose regultion in ptients with type 2 dibetes mellitus. J Int Med Res. 2014;42: Brownlee M. The pthobiology of dibetic complictions: unifying mechnism. Dibetes. 2005;54: Nuseef WM. Dignostic ssys for myeloperoxidse nd myeloperoxidse deficiency. Methods Mol Biol. 2014;1124: Lnz F. Clinicl mnifesttion of myeloperoxidse deficiency-review. J Mol Med (Berl). 1998;76: Mrchetti C, Ptrirc P, Solero GP, Brlle FE, Romno M. Genetic chrcteriztion of myeloperoxidse deficiency in Itly. Hum Mutt. 2004;23: Dinuer MC, Newburger PE. The phgocyte system nd disorders of grnulopoiesis nd grnulocyte function. In: Orkin SH, Nthn DG, Ginsburg D, Look AT, Fisher DE, Lux SE, editors. Nthn nd Oski s Hemtology of Infncy nd Childhood. Phildelphi: Sunders-Elsevier; p Abdi Liie Z, Soudbkhsh A, Atrod L, Toogeh GH, Nkhjvni M, Mousvipnh P, et l. Myeloperoxidse deficiency in neutrophils of dibetic ptients with nd without infectious disese. Tehrn Univ Med J. 2008;65(12): Sto N, Shimizu H, Suw K, Shimomur Y, Kobyshi I, Mori M. MPO ctivity nd genertion of ctive O 2 species in leukocytes from poorly controlled dibetic ptients. Dibetes Cre. 1992;15: Gorudko IV, Kostevich AV, Sokolov AV, Konsttinov EÉ, Tspev NL, Mironov EV, et l. Incresed myelopepoxidse ctivity is risk fctor for ishemic hert disese in ptients with dibetes mellitus. Biomed Khim. 2012;58: Americn Dibetes Assocition. Executive summry: stndrds of medicl cre in dibetes Dibetes Cre. 2014;37(Suppl 1):S Kithr M, Simonin Y, Eyre HJ. Neutrophil myeloperoxidse: simple, reproducible technique to determine ctivity. J Lb Clin Med. 1979;93: Nit C, Bl C, Porojn M, Hncu N. Fenofibrte improves endothelil function nd plsm myeloperoxidse in ptients with type 2 dibetes

7 Pge 7 of 7 mellitus: n open-lbel interventionl study. Dibetol Metb Syndr. 2014;6: Uchimur K, Ngsk A, Hyshi R, Mkino M, Ngt M, Kkizw H, et l. Chnges in superoxide dismutse ctivities nd concentrtions nd myeloperoxidse ctivities in leukocytes from ptients with dibetes mellitus. J Dibetes Complictions. 1999;13: Moldovenu E, Tnsenu C, Tnsenu S, Kosk T, Mne G, Mrt DS, et l. Plsm mrkers of endothelil dysfunction in type 2 dibetics. Eur J Intern Med. 2006;17: Gómez-Grcí A, Rodríguez MR, Gómez-Alonso C, Ocho DYR, Alvrez- Aguilr C. Myeloperoxidse is ssocited with insulin resistnce nd inflmmtion in overweight subjects with first-degree reltives with type 2 dibetes mellitus. Dibetes Metb J. 2015;39: de Souz Ferreir C, Arújo TH, Ângelo ML, Penncchi PC, Okd SS, de Arújo Pul FB, et l. Neutrophil dysfunction induced by hyperglycemi: modultion of myeloperoxidse ctivity. Cell Biochem Funct. 2012;30: Sito Y, Tkhshi I, Iwne K, Okubo N, Nishimur M, Mtsuzk M, et l. The influence of blood glucose on neutrophil function in individuls without dibetes. Luminescence. 2013;28: Alb-Loureiro TC, Munhoz CD, Mrtins JO, Cerchiro GA, Scvone C, Curi R, et l. Neutrophil function nd metbolism in individuls with dibetes mellitus (Review). Brz J Med Biol Res. 2007;40: Gllcher SJ, Thomson G, Frser WD, Fisher BM, Gemmell CG, McCuish AC. Neutrophil bctericidl function in dibetes mellitus: evidence for ssocition with blood glucose control. Dibet Med. 1995;12: Ayilvrpu S, Kntrci A, Fredmn G, Turkoglu O, Omori K, Liu H, Iwt T, Ygi M, Hsturk H, Vn Dyke TE. Dibetes-induced oxidtive stress is medited by C 2+ independent phospholipse A2 in neutrophils. J Immunol. 2010;184: Tong PC, Lee KF, So WY, et l. White blood cell count is ssocited with mcro nd microvsculr complictions in Chinese ptients with type 2 dibetes. Dibetes Cre. 2004;27: Lou M, Luo P, Tng R, Peng Y, Yu S, Hung W, He L. Reltionship between neutrophil-lymphocyte rtio nd insulin resistnce in newly dignosed type 2 dibetes mellitus ptients. BMC Endocr Disord. 2015;2(15):9. doi: /s Muchov J, Liptkov A, Orszghov Z, Griov I, Tison P, Crsky J, Durckov Z. Antioxidnt systems in polymorphonucler leucocytes of type 2 dibetes mellitus. Dibet Med. 1999;16: Goh SY, Cooper ME. Clinicl review: the role of dvnced glyction end products in progression nd complictions of dibetes. J Clin Endocrinol Metb. 2008;93: Brownlee M. Biochemistry nd moleculr cell biology of dibetic complictions. Nture. 2001;414: Iked K, Higshi T, Sno H, Jinnouchi Y, Yoshid M, Arki T, et l. N ε (crboxymethyl) lysine protein dduct is mjor immunologicl epitope in proteins modified with dvnced glyction end products of the Millrd rection. Biochemistry. 1996;35: Klebnoff SJ. Myeloperoxidse: friend nd foe (Review). J Leukoc Biol. 2005;77: Kutter D, Devquet P, Vnderstocken G, Pulus JM, Mrchl V, Gothot A. Consequences of totl nd subtotl myeloperoxidse deficiency: risk or benefit? Act Hemtol. 2000;1041: Tng WH, Wu Y, Nicholls SJ, Hzen SL. Plsm myeloperoxidse predicts incident crdiovsculr risks in stble ptients undergoing medicl mngement for coronry rtery disese. Clin Chem. 2011;57: Golubinsky V, Brndt-Elisson U, Gn LM, Kjerrulf M, Nilsson H. Endothelil function in mouse model of myeloperoxidse deficiency. Biomed Res Int. 2014;2014: Jy D, Hitomi H, Griendling KK. Oxidtive stress nd dibetic crdiovsculr complictions. Free Rdic Biol Med. 2006;40: Nsr CE, Hoogwerf BJ, Fimn C, Reddy SS. United Kingdom Prospective Dibetes Study (UKPDS). Effects of glucose nd blood pressure control on complictions of type 2 dibetes mellitus. Cleve Clin J Med. 1999;66: Zhng R, Brennn ML, Fu X, Aviles RJ, Perce GL, Penn MS, et l. Assocition between myeloperoxidse levels nd risk of coronry rtery disese. JAMA. 2001;286: Submit your next mnuscript to BioMed Centrl nd we will help you t every step: We ccept pre-submission inquiries Our selector tool helps you to find the most relevnt journl We provide round the clock customer support Convenient online submission Thorough peer review Inclusion in PubMed nd ll mjor indexing services Mximum visibility for your reserch Submit your mnuscript t