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*E-mail: nunez500412@gmail.com
Abstract Resumen
Context: A clinical study was conducted in a cohort of human subjects Contexto: Se realizó un estudio clínico en una cohorte de sujetos
from 60 to 75 years of age in order to assess the correlation between humanos entre 60 y 75 años de edad para determinar la correlación entre
disease diagnosis and oxidative stress (OS) damage in diabetes mellitus diagnóstico de la enfermedad y daño por estrés oxidativo (EO) en
(DM) and arterial hypertension (AHT). diabetes mellitus (DM) e hipertensión arterial (HTA).
Aims: To provide a sound basis to design a clinical protocol for Objetivos: Proporcionar una base sólida para el diseño de un protocolo
antioxidant adjuvant therapy for DM and AHT. clínico para la terapia adyuvante antioxidante en DM e HTA.
Methods: The values of the OS index were determined from the Métodos: Se determinaron los valores del índice de EO a partir de las
experimental values of biomarkers in erythrocyte lysates obtained from determinaciones experimentales de niveles de antioxidantes totales,
human blood samples including total antioxidant levels, specific biomarcadores específicos del daño oxidativo y actividades de enzimas
biomarkers of oxidative damage, and antioxidant enzyme activities. antioxidantes en lisados de eritrocitos obtenidos de muestras de sangre.
Three study groups were formed: Group I: DM patients (110 subjects), Se conformaron tres grupos de estudio: Grupo I; pacientes DM (110
group II: AHT patients (112 subjects); and Control group: Healthy sujetos), grupo II; pacientes HTA (112 sujetos), y grupo III; voluntarios
volunteers (123 subjects). The data of all groups were analyzed using sanos (123 sujetos). Los datos de los grupos se analizaron con el
SPSS 9.0 software. A nonparametric Friedman test was used to compare programa SPSS 9.0. Se utilizó la prueba no paramétrica de Friedman
several related subgroups, and changes within the groups and para comparar entre subgrupos, y los cambios entre grupos se
subgroups were tested using the Wilcoxon paired test. A Mann-Whitney analizaron mediante la prueba de pares de Wilcoxon. Las diferencias
U test was used to estimate significant differences (p<0.05) between the significativas entre subgrupos (p<0.05) se estimaron con la prueba de
subgroups. Whitney U.
Results: Severe OS was observed in subgroup IIc (AHT III), moderate OS Resultados: Se observó EO severo en el subgrupo IIc (HTA III), EO
was observed in subgroups Ib (DM II) and IIb (AHT II), mild OS was moderado en los subgrupos Ib (DM II) y IIb (HTA II), EO leve en el
observed in subgroup Ia (DM I), and no OS was observed in subgroup subgrupo Ia (DM I), y no se observó EO en el subgrupo IIa (HTA I).
IIa (AHT I). Conclusiones: Los resultados demuestran el posible diseño de protocolos
Conclusions: The results support the possible design of clinical trial de ensayos clínicos para incrementar la eficacia de las terapias estándar
protocols for adjuvant antioxidant therapy in order to increase the de la HTA II y III, y DM II, con el uso de terapias antioxidantes
efficacy of standard therapies for AHT II and III and for DM II. adyuvantes, las que no son recomendables para DM I y HTA I, debido a
Antioxidant therapies for DM I and AHT I are not recommended due to que el EO es leve o no existente, respectivamente.
the presence of only mild OS or no OS, respectively.
Keywords: antioxidant therapy; diabetes mellitus; hypertension; Palabras Clave: biomarcadores redox; estrés oxidativo; diabetes mellitus;
oxidative stress; redox biomarkers. hipertensión arterial; terapia antioxidante.
ARTICLE INFO
Received: January 16, 2019.
Received in revised form: February 21, 2019.
Accepted: February 22, 2019.
Available Online: February 26, 2019.
Declaration of interests: The authors declare no conflict of interest.
Funding: This work was supported by the Ministry of Higher Education, Science and Technology (MESCYT), Dominican Republic, Project FONDOCYT 2012-
2013-2A1-58, and by the National Evangelic University (UNEV), Santo Domingo, Dominican Republic, Project UNEV 2013/004.
_____________________________________
Núñez Sellés et al. Oxidative stress index in arterial hypertension and diabetes mellitus
The sample size that was required to obtain sta- II FPG > 126 mg/dL
tistically significant results was determined using DM: Diabetes mellitus; FPG: Fasting plasma glucose.
validated software (Faul et al., 2007) considering
the incidence of DM and AHT in various age Subjects were evaluated clinically by a physi-
groups in the Santo Domingo population. Three cian, DM and AHT diagnoses were confirmed
groups and 10 subgroups were formed during the based on previous analysis, blood samples (100
study to obtain a significance level (α) = 0.05, po- μL) were collected and data were recorded in the
tency (1-β) = 0.2, standard deviation (σ) = 2, and Data Collection Logbook (DCL) in duplicate using
detection difference (d) = 1. A total of 345 subjects a code. Lists with the personal data of included or
were recruited. excluded subjects were submitted by the physi-
cians to the Principal Researcher.
Inclusion criteria were as follows: i) patients of
both sexes from 60 to 75 years of age with a con- Blood samples
firmed diagnosis of DM (types I and II) and/or
Blood samples (100 μL) were collected by finger
AHT (degrees I, II, and III) from Santo Domingo
puncture with a sterilized lancet and transferred
Province, ii) patients with disease presence for at
into an Eppendorf vial containing 2 mL saline.
least 2 years or more iii) patients who were non-
Samples were maintained at 8°C until transporta-
smokers, iv) patients who did not regularly con-
tion to the analytical laboratory within the next 24
sume alcoholic drinks or stupefacient drugs, and
h. Samples were centrifuged at 3,000 xg. The su-
v) patients who did not consume of antioxidants
pernatant was discarded, and 2 mL of double-
during the last 12 months. Exclusion criteria were
distilled water was added to the erythrocyte frac-
as follows: i) other chronic degenerative diseases,
tion to induce an osmotic shock and the conse-
and ii) participation in other clinical studies.
quence lysis. Fractions of erythrocyte (RBC) lysate
AHT patients were classified according to the (200 μL) were collected with a micropipette, trans-
Clinical Practice Guides for Arterial Hypertension ferred into Eppendorf vials (1 mL), coded, and
Treatment (Mancia et al., 2007) as shown in Table 1. maintained at -10°C in a vertical freezer until pro-
DM patients were classified according to Diabetes cessing within the next 60 days.
Management Guidelines (American Diabetes Associ-
ation, 2016) as shown in Table 2. Laboratory methods
Total antioxidant status (TAS) sorbance was determined at 370-375 nm. HCl,
TAS values were determined based on the ki- ethyl acetate, and ethanol were purchased from JT
netics of formation of free radicals in the reaction Baker, USA; DNPH, TCA, guanidine.HCl, and egg
albumin were purchased from Sigma-Aldrich,
with ABTS+) -2,2'-azinobis-(3-ethylbenzothiazo-
line-6-sulfonic acid radical cation- (Sigma Aldrich, Missouri, USA.
Missouri, USA) at 600 nm according to Ozcan
Sulfhydryl groups (SG)
(2004).
SG values were determined according to Sedlak
Trolox equivalent antioxidant capacity (TEAC) and Lindsay (1968). RBC lysates were dissolved in
1 mL potassium phosphate buffer (50 mM, pH =
TEAC values were determined by the modified
method of Blois (1958). RBC lysates or Trolox were 7.5) and centrifuged. The supernatant was trans-
ferred into a vial containing Ellman’s reagent and
incubated with sodium acetate buffer, 40 mM, pH
= 5.5, and 1 mL ethanol for 1 h at 37°C. Then, 1 mL absorbance was determined at 600 nm. The same
procedure was used in the case of the nonoxidized
of 100 μM 1,1-diphenyl-2-picrylhydrazyl (DPPH)
control protein sample (egg albumin). All reagents
was added after incubation to an aliquot of the
were purchased from JT Baker, USA, except egg
incubated sample and absorbance was determined
albumin from Sigma-Aldrich, Missouri, USA.
at 517 nm in the aliquots with and without DPPH.
Trolox and DPPH were purchased from Sigma-
Superoxide dismutase (SOD)
Aldrich, Missouri, USA; other reagents from JT
Baker, USA. SOD activity was determined according to Mes-
tro and McDonald (1986). RBC lysates were mixed
Malondialdehyde (MDA) with 2.8 mL buffer containing 100 μL 05 M Tris
[tris(hydroxymethyl)aminomethane], pH = 8.2, in
MDA concentrations were determined accord-
distilled water and 50 μL of EDTA solution (1
ing to Esterbauer and Cheeseman (1990). RBC ly-
mM). Then, 50 μL pyrogallol solution (0.124 M)
sates were analyzed using an LPO-586 kit (Calbio-
was added and absorbance values were deter-
chem, La Jolla, CA, USA). The production of a sta-
mined at 420 nm after 10 s and then every 10 s for
ble chromophore after 40 min incubation at 45°C
1 min (six absorbance values). The same procedure
was measured at 586 nm. Freshly prepared solu-
tions of MDA bis[dimethylacetal] were used as a was used in the case of a blank sample without
standard and were assayed under identical condi- blood. Absorbance differences between the sample
tions. MDA was purchased from Sigma-Aldrich, and the blank were calculated and SOD activity
Missouri, USA. was determined. All reagents were purchased
from Sigma-Aldrich, Missouri, USA.
Carbonyl groups (CG)
Catalase (CAT)
CG values were determined according to Oliver
et al. (1987). RBC lysates were suspended in 1 mL CAT activity was determined according to
hydrochloric acid (2 M) and 1 mL 2,4- Haining and Legan (1972). RBC lysates were
dinitrophenylhydrazine (0.2 %) and incubated mixed with 0.9 mL potassium phosphate buffer
with stirring at 37°C for 1 h. Then, 1 mL 10 % tri- (pH = 7) and 500 μL buffered hydrogen peroxide
solution (0.05 M) in a quartz cell. Absorbance at
chloroacetic acid (TCA) was added and the precip-
240 nm was determined at 10 and 70 s. Blank ab-
itate was extracted with 10 mL ethanol:ethyl ace-
sorbance was determined using 1.5 mL buffer so-
tate (1:1). The solid was reprecipitated with TCA
lution. Absorbance differences between the sample
(10 %). Protein control sample (egg albumin) was
and the blank were calculated and CAT activity
dissolved in 1 mL guanidine hydrochloride in po-
was determined. All reagents were purchased
tassium phosphate buffer (20 mM, pH = 6.5). The
from Sigma-Aldrich, Missouri, USA.
sample and the control were centrifuged, and ab-
Table 4. Mean age and gender data from the cohort population study.
OSD ± SD
Gender Age range (years old) Term MiA Term MiB Term MiC
(p<0.05)
OSI ± SD
Gender Age range (years old) Term MiA Term MiB Term MiC
(p<0.05)
OSI ± SD
Gender Age range (years old) Term MiA Term MiB Term MiC
(p < 0.05)
≥65 y 40.6 ± 2.4a 46.0 ± 2.4ab 43.1 ± 2.6ab 69.1 ± 1.8 abcde
Different letters mean statistical difference (p<0.05) between the same series. OSI: Oxidative stress index. MiA = A1 (PP) + A2 (TAS) + A3 (TEAC).
MiB = B1 (MDA) + B2 (carbonyl groups) + B3 (sulfhydryl groups). MiC = C1 (SOD) + C2 (CAT) + C3 (GPx). PP: Peroxidation potential, TAS: Total
antioxidant status, TEAC: Trolox equivalent antioxidant capacity, MDA: Malondialdehyde, SOD: Superoxide dismutase, CAT: Catalase, GPx:
Glutathione peroxidase.
Table 9. Determination of the oxidative damage degree in groups I and II according to the oxidative stress index.
OSI
Age range
Gender Subgroup Ia Subgroup Ib Subgroup IIa Subgroup IIb Subgroup IIc
(years old)
(DM type I) (DM type II) (AHT degree I) (AHT degree II) (AHT degree III)
Male 60 - 64 y 33.0 ± 6.8 53.4 ± 3.5b -5.8 ± 12.4 56.6 ± 5.3 abcd 67.9 ± 2.3 abcd
≥65 y 29.0 ± 8.6 46.1 ± 7.2c -9.7 ± 12.3 51.8 ± 4.5 abcd 66.5 ± 1.2 abcd
≥65 y 26.1 ± 8.8a 43.9 ± 6.1abc 6.4 ± 7.8 65.9 ± 2.0 abcd 69.1 ± 1.8 abcde
extrapolated to other age populations or geo- implies a useful tool to demonstrate the role of or
graphical origins with unknown baseline data for relationship between these biomarkers and the
the OS biomarkers used. pathophysiology of a disease. Measuring the base-
line values in naive situations in the case of a sin-
The aim to demonstrate how a single measure
gle test of biomarkers has resulted in a valuable
of any type of OS biomarker reflects the responses
information in some cases (Lee et al., 2007).
of the body to external and/or internal factors and
hypertension, OS promotes endothelial dysfunc- II and III and DM II may interfere with the OS–
tion, vascular remodeling, and inflammation, lead- mediated signaling pathways and is a safe and
ing to vascular damage. Although experimental promising recommendation.
evidence indicates a causative role for OS in hy-
The present study heads its conclusions to bet
pertension, the human data are less convincing.
for a correct application of effective antioxidant
This may be related, in part, to suboptimal meth-
therapy may target the pathophysiogical changes
ods used to assess the redox state (Montezano et
involved in the development of the disease and
al., 2015). The results of subgroups IIb and IIc indi-
may prevent the complications or lower their oc-
cated the presence of moderate and severe ODD,
currence and severity. The favorable effects of an-
respectively (Table 9), signifying that an increase
tioxidants given as a supplement in DM, even if
in OSI may lead to higher degrees of hypertension.
specific antioxidant needs have not been deter-
The question whether antioxidant therapy would
mined, may significantly improve prognoses and
be recommended in these subgroups remains un-
were reported to prevent the complications (Testa
answered. Most clinical trials on the use of the
et al., 2016).
antioxidant vitamins C, E, and A (β-carotene) or
their combinations to lower blood pressure have Thus, complications of DM II and/or AHT II-III
failed (Heart Protection Study Collaborative and OS related to these diseases may be prevented
Group, 2002), which has limited support for the or ameliorated by the opportune application of
use of antioxidants as adjuvants as antihyperten- antioxidant adjuvant therapy and may benefit by
sive therapies. Nevertheless, the use of antioxidant the selection of appropriate and specific com-
cocktails, considering the different targets of the pounds.
antioxidant therapy, has not been reported. At
present, it is not clear what mechanisms should be CONCLUSIONS
targeted and what types of compounds should be Our experimental evidence suggests that an
used in order to increase the efficacy of standard appropriate OS diagnosis can optimize an inter-
antihypertensive treatment with antioxidants vention with an antioxidant cocktail. OSI determi-
(Münzel et al., 2010). nation in DM II and AHT II and III within a well-
A number of studies have reported the benefi- designed clinical protocol will provide the neces-
cial effect of dietary antioxidants in hypertension sary support to optimize the use of antioxidant
and cardiovascular risk; however, the lack of effi- therapies in these diseases to increase efficacy and
cacy in lowering the blood pressure in the cases of to decrease comorbidities in combination within
AHT has been highlighted in other scenarios the standard therapies currently used to treat these
(Yusuf et al., 2000). Nonetheless, other negative diseases. Currently, despite considerable contro-
factors associated with chronic diseases are in- versy, increasing use of antioxidants in combina-
volved in their pathogeneses, and OS plays a ma- tion with the standardized antihypertensive or
jor role; hence, AHT and DM, in particular, may antidiabetic therapies appears to be the most effec-
benefit from balanced diets containing antioxidant tive means to improve the prognosis of DM II and
components (Poprac et al., 2017). It should be not- AHT II and III due to control over the molecular
ed that more than one (three or four) antioxidant mechanisms involved in the regulation of vascular
components are expected to be combined in a for- function, metabolic balance and redox states by
mulation to provide a real benefit in the adjuvant proper management with specific antioxidants
treatment of AHT and DM (Wu et al., 2014). This (Sorriento et al., 2018). The application of antioxi-
may be because balanced diets enriched with a dant therapies may be useful to combat complica-
wide variety of fruits are advantageous in improv- tions or deteriorations due to physiological imbal-
ing the levels of blood sugar and blood pressure in ances in DM and AHT.
certain cases; thus, complimentary therapy against The results of the present trial support second-
the endothelial dysfunctions associated with AHT ary prevention and indicate that the use of antiox-
http://jppres.com/jppres J Pharm Pharmacogn Res (2019) 7(2): 112
Núñez Sellés et al. Oxidative stress index in arterial hypertension and diabetes mellitus
idant therapies in DM I and AHT I should not be macrovascular complications: Avenues for a mechanistic-
recommended in patients who have demonstrated based therapeutic approach. Curr Diab Rev 7: 313–324.
mild antioxidant responses to their current treat- Frijhoff J, Winyard PG, Zarkovic N, Davies SS, Stocker R,
Cheng D, Knight AR, Taylor EL, Oettrich J, Ruskovska T,
ment strategies. Gasparovic AC, Cuadrado A, Weber D, Poulsen HE,
Grune T, Schmidt HHW, Ghezzi P (2015) Clinical
CONFLICT OF INTEREST relevance of biomarkers of oxidative stress. Antioxid
Redox Signal 23: 1144–1170.
The authors declare no conflict of interest.
Gil L, Martínez G, González I, Tarinas A, Álvarez A, Giuliani
A, Molina R, Tápanes R, Pérez J, León OS (2003)
ACKNOWLEDGMENTS Contribution to characterization of oxidative stress in
HIV/AIDS patients. Pharmacol Res 47: 217–224.
This work was supported by the Ministry of Higher Edu-
cation, Science and Technology (MESCYT), Dominican Re- Haining JL, Legan JS (1972) Improved assay for catalase based
public, Project FONDOCYT 2012-2013-2A1-58, and by the upon steady-state substrate concentration. Anal Biochem
National Evangelic University (UNEV), Santo Domingo, Do- 45: 469–479.
minican Republic, Project UNEV 2013/004. The authors thank Heart Protection Study Collaborative Group (2002)
R. Silverio, D.A. Terrero, and M. Beras for their assistance in MRC/BHF Heart Protection Study of antioxidant vitamin
recruiting patients in the hospitals. D. Jackson, L. Kelly, J.M. supplementation in 20,536 high-risk individuals: A
Jimenez, and L. Perez for their assistance in recruiting healthy randomised placebo-controlled trial. Lancet 360: 23–33.
volunteers. M. Butler and P. Santana for the logistic support Lee SA, Kallianpur A, Xiang YB, Wen W, Cai Q, Liu Q, Shu
by the Faculty of Health Sciences, UNEV. XO (2007) Intra-individual variation of plasma adipokine
levels and utility of single measurement of these
SUPPLEMENTARY INFORMATION biomarkers in population-based studies. Cancer
Epidemiol Biomark Prev 16: 2464–2470.
Clinical Trial Approval by the National Regulatory Com-
Lodovicia M, Giovannellia L, Pitozzia V, Bigaglia E, Bardinib
mission. Protocol No. 008-2016. (www.conabios.gob.do).
G, Rotellab CM (2008) Oxidative DNA damage and
plasma antioxidant capacity in type 2 diabetic patients
REFERENCES with good and poor glycaemic control. Mutat Res 638:
98–102.
Ahmad KA, Yuan Yuan D, Nawaz W, Ze H, Zhuo CX, Talal B,
Qilong D (2017) Antioxidant therapy for management of Mancia G, Backer GD, Dominiczak A, Cifkova R, Fagard R.,
oxidative stress induced hypertension. Free Radic Res 51: Germano G, Narkiewicz K, Ruilope L, Rynkiewicz A,
428–438. Schmieder RE, Struijker Boudier HAJ, Zanchetti A (2007)
Guías de práctica clínica para el tratamiento de la
American Diabetes Association (2016) Standards of medical
hipertensión arterial 2007. Rev Esp Cardiol 60(9): 968.e1-
care in diabetes-2016. Diabetes Care 39: S1–S106.
e94.
Block G, Dietrich M, Norkus EP, Morrow JD, Hudes M, Caan
Mañon Rossi W, Garrido G, Nuñez Selles AJ (2016)
B, Packer L (2002) Factors associated with oxidative stress
Biomarkers of oxidative stress in antioxidant therapy. J
in human populations. Am J Epidemiol 156:274–285.
Pharm Pharmacogn Res 4: 62–83.
Blois M (1958) Antioxidant determinations by the use of a
Marrocco I, Altieri F, Peluso I (2017) Measurement and clinical
stable free radical. Nature 181: 1199–2000.
significance of biomarkers of oxidative stress in humans.
Escobar Aedo AL, Pérez Ortega Y, Vera González M, Oxid Med Cell Long 2017: 6501046.
Peñaranda Guzmán O, Álvarez León A (2011) Effect of
Mestro RF, McDonald W (1986) Oxidative enzymes in tissue
Vimang supplementation on oxidative stress markers in
homogenates in Greenwald RA (ed) CRC Handbook for
young patients with diabetes mellitus type 1. Rev Cub
Oxygen Radical Research. CRC Press, Boca Raton FL. p.
Endocrinol 22: 103–117.
291–296.
Esterbauer H, Cheeseman KH (1990) Determination of
Mondal LK, Bhaduri G, Bhattacharya B (2018) Biochemical
aldehydic lipid peroxidation products: malonaldehyde
scenario behind initiation of diabetic retinopathy in type
and 4-hydroxynonenal. Methods Enzymol 186: 407–421.
2 diabetes mellitus. Indian J Ophthalmol 66:535–540.
Faul F, Erdfelder E, Lang AG, Buchner A (2007) GPower 3: A
Montezano AC, Dulak-Lis M, Tsiropoulou S, Harvey A,
flexible statistical power analysis for the social,
Briones AM, Touyz RM (2015) Oxidative stress and
behavioral and biomedical sciences. Behav Res Meth 39:
human hypertension: vascular mechanisms, biomarkers,
175–191.
and novel therapies. Can J Cardiol 31: 631–641.
Folli F, Corradi D, Fanti P, Davalli A, Paez A, Giaccari A,
Münzel T, Gori T, Bruno RM, Taddei S (2010) Is oxidative
Muscogiuri G (2011) The role of oxidative stress in the
stress a therapeutic target in cardiovascular disease? Eur
pathogenesis of type 2 diabetes mellitus micro-and
Heart J 31: 2741–2748.
http://jppres.com/jppres J Pharm Pharmacogn Res (2019) 7(2): 113
Núñez Sellés et al. Oxidative stress index in arterial hypertension and diabetes mellitus
National Office of Statistics (2012) Dominican Republic: Vital oxidative protein damage and antioxidant enzyme
Statistics 2010. IX National Census of Population & activities in plasma of patients with different degrees of
Housing. Santo Domingo, Dominican Republic. essential hypertension. J Hum Hypertens 20: 149–155.
Nuñez Sellés AJ, Martínez G, Mañón Rossi W (2017) Method Sinha N, Dabla PK (2015) Oxidative stress and antioxidants in
for determining oxidative stress index in patients with hypertension-a current review. Curr Hypertens Rev 11:
diabetes mellitus and arterial hypertension. Oficina 132–142.
Nacional de la Propiedad Industrial (ONAPI), República Society of Cardiology, Dominican Republic (2012) Study of
Dominicana. 2017; P2017–54, March 14. Cardiovascular Risk Factors and Metabolic Syndrome II.
Oliver CN, Ahn B, Moerman EJ, Goldstein S, Stadtman ER (EFRICARD II -in Spanish-). Santo Domingo, Dominican
(1987) Age related changes in oxidised protein. J Biol Republic.
Chem 262: 5488–5491. Sorriento D, De Luca N, Trimarco B, Iaccarino G (2018) The
Ozcan E (2004) A novel automated direct measurement antioxidant therapy: New insights in the treatment of
method for total antioxidant capacity using a new hypertension. Front Physiol 9: 528.
generation, more stable ABTS cation. Clin Biochem 37: Stephens JW, Khanolkar MP, Bain SC (2009) The biological
277–285. relevance and measurement of plasma markers of
Ozdemirler G, Mehmetcik G, Oztezcan S, Toker G, Sivas A, oxidative stress in diabetes and cardiovascular disease.
Uysal M (1995) Peroxidation potential and antioxidant Atheroscl 202: 321–329.
activity of serum in patients with diabetes mellitus and Testa R, Bonfigli AR, Genovese S, De Nigris V, Ceriello A
myocardial infarction. Horm Metab Res 27: 194–196. (2016) The possible role of flavonoids in the prevention of
Peruzzi C, Nascimento S, Gauer B, Nardi J, Sauer E, Göethel diabetic complications. Nutrients 8: 310.
G, Cestonaro L, Fão N, Cattani S, Paim C, Souza J, Wu Y, Tang L, Chen B (2014) Oxidative stress: Implications
Gnoatto D, Garcia SC (2019) Inflammatory and oxidative for the development of diabetic retinopathy and
stress biomarkers at protein and molecular levels in antioxidant therapeutic perspectives. Oxid Med Cell
workers occupationally exposed to crystalline silica. Long 2014: 752387.
Environ Sci Pollut Res Int 26(2): 1394–1405.
Yao EH, Fukuda N, Matsumoto T, Kobayashi N, Katakawa M,
Poprac P, Jomova K, Simunkova M, Kollar V, Rhodes CJ, Yamamoto C, Tsunemi A, Suzuki R, Ueno T, Matsumoto
Valko M (2017) Targeting free radicals in oxidative stress- K (2007) Losartan improves the impaired function of
related human diseases. Tr Pharmacol Sci 38: 592–607. endothelial progenitor cells in hypertension via an
Profumo E, Buttari B, Saso L, Rigano R (2014) Pleiotropic antioxidant effect. Hypertens Res 30: 1119–1128.
effects of statins in atherosclerotic disease: focus on the Yoshida Y, Saito Y, Hayakawa M, Habuchi Y, Imai Y, Sawai Y,
antioxidant activity of atorvastatin. Curr Top Med Chem Niki E (2007) Levels of lipid peroxidation in human
14: 2542–2551. plasma and erythrocytes: Comparison between fatty
Sas K, Szabó E, Vécsei L (2018) Mitochondria, oxidative stress acids and cholesterol. Lipids 42: 439–449.
and the kynurenine system, with a focus on ageing and Yusuf S, Sleight P, Pogue J, Bosch J, Davies R, Dagenais G
neuroprotection. Molecules 23: pii: E191. (2000) Effects of an angiotensin-converting-enzyme,
Sedlak J, Lindsay RH (1968) Estimation of total protein bond ramipril, on cardiovascular events in high-risk patients.
and non-protein sulfhydryl group with Ellman’s reagent. N Engl J Med 342: 145–153.
Anal Biochem 25: 192–205. Zilmer M, Soomets U, Rehema A, Langel Ü (2005) The
Simic DV, Mimic-Oka J, Pljesa-Ercegovac M, Savic-Radojevic glutathione system as an attractive therapeutic target.
A, Opacic M, Matic D, Simic T (2006) Byproducts of Drug Design Rev 22: 121–127.
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AUTHOR CONTRIBUTION:
Contribution Núñez Sellés AJ Mañon Rossi WI Núñez Musa R Guillén Marmolejos R Martínez-Sánchez G
Concepts or ideas x x
Design x
Literature search x x x
Experimental studies x x x x x
Clinical trials x x x x
Data acquisition x x x x
Data analysis x x x
Statistical analysis x
Manuscript preparation x x x x x
Manuscript editing x x x
Manuscript review x x x x x
Citation Format: Núñez Sellés AJ, Mañon Rossi WI, Núñez Musa R, Guillén Marmolejos R, Martínez-Sánchez G (2019) The oxidative stress
index in human population with arterial hypertension and diabetes mellitus. J Pharm Pharmacogn Res 7(2): 103–115.