<i>FOXE1</i> polymorphisms and chronic exposure to nitrates in drinking water cause metabolic dysfunction, thyroid abnormalities, and genotoxic damage in women

Gandarilla-Esparza, Diana Dennys; Calleros-Rincón, Esperanza Yasmin; Macias, Hortensia Moreno; González-Delgado, María Fernanda; Vargas, Gonzalo García; Sustaita, Jaime Duarte; González-Zamora, Alberto; Ríos-Sánchez, Efraín; Pérez-Morales, Rebeca

doi:10.1590/1678-4685-GMB-2021-0020

Diana Dennys Gandarilla-Esparza

Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Gómez Palacio, Durango, México.

Esperanza Yasmin Calleros-Rincón

Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Gómez Palacio, Durango, México.

Hortensia Moreno Macias

Universidad Autónoma Metropolitana, División CSH de la Unidad Iztapalapa, Departamento de Economía, Vicentina, Ciudad de México, México.

María Fernanda González-Delgado

Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Gómez Palacio, Durango, México.

Gonzalo García Vargas

Universidad Juárez del Estado de Durango, Facultad de Ciencias de la Salud, Departamento de Investigación. Gómez Palacio, Durango, México.

Jaime Duarte Sustaita

Universidad Juárez del Estado de Durango, Facultad de Ciencias de la Salud, Departamento de Investigación. Gómez Palacio, Durango, México.

Alberto González-Zamora

Universidad Juárez del Estado de Durango, Facultad de Ciencias Biológicas, Laboratorio de Biología Evolutiva, Gómez Palacio, Durango, México.

Efraín Ríos-Sánchez

Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Gómez Palacio, Durango, México.

Rebeca Pérez-Morales

Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Gómez Palacio, Durango, México.

http://orcid.org/0000-0003-1102-4553

Send correspondence to
Rebeca Pérez-Morales. Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Av. Artículo, 123, Fracc., Filadelfia, C.P. 35010. Gómez Palacio, Durango, México. E-mail: rebecapms@ujed.mx

Associate Editor: Mara Hutz

Conflict of Interest

The authors declare that there is no conflict of interest that could be perceived as prejudicial to the impartiality of the reported research.

Author contributions

DDGE performed experimental work and help writing the results; EYCR participated in the design of the study and sampling; HMM performed the multivariate models and statistical analyses; MFGD participated in the analysis of biochemical parameters and interpretation of results; GGGV and JDS performed the analysis in water and urine samples; AGZ and ERS participated in curating the database and conducted the statistical analyses, edited images and participated in writing and discussion of the manuscript; RPM designed the study, participated in the implementation of techniques and wrote the manuscript. All authors read and approved the final version.

Levels of Nitrate mg/L by years (µ ± SD)
Communities		21 marzo Sapioriz Cd. Juárez	San Jacinto Juan E. García,	La Loma Nazareno
2010 - 2012^£ 2013 - 2014^¥ 2015 - 2017^§		3.2 3.8 ± 3.2 4.7 ± 3.3	18.14 24.1 ± 3.2 32.1 ± 3.7	36.5 47.6 ± 7 56.9 ± 14.7
Daily consumption of drinking water (L) (µ±SD)		2.08 ± 1.03	1.91 ± 0.82	2.4 ± 0.95
Parameter	All women n=420	Low level n=139	Medium level n=171	High level n=110	P-value
Age (years)	36 (32-40)	36 (33-40)	35 (30-41)	37 (33-40)	0.4023^*
BMI (kg/m²) < 24.9 25-29.9 ≥ 30	28.5 (25.5-32.4) 88 (21%) 173 (41%) 159 (38%)	29.3 (26.5-32.9) 17 (12% 62 (45%) 60 (43%)	28.5 (24.7-33.3)^a 45 (26%) 56 (33%) 70 (41%)	27.9 (25.3-30)^b 26 (24%) 55 (50%) 29 (26%)	0.0433^* 0.006^ 0.018^ 0.041^**
Education < high school > high school	359 (85%) 61 (15%)	125 (90%) 14 (10%)	135 (79%) 36 (21%)	99 (90%) 11 (10%)	0.007^**
Occupation Homemaker Seasonal agriculture	399 (95%) 21 (5%)	132 (95%) 7 (5%)	159 (93%) 12 (7%)	106 (97%) 4 (3%)	0.40^*
Drinking water Well Purified	335 (80%) 85 (20%)	118 (85%) 21 (15%)	124 (73%) 47 (27%)	93 (85%) 17 (15%)	0.009^**
Smoking current	28 (6%)	11 (7%)	14 (8%)	3 (2%)	0.138^**
Alcohol consumption	57 (13%)	18 (12%)	28 (16%)	11 (10%)	0.293^**

Parameters	All women n=420	Low n=139	Medium n=171	High n=110	Reference rank	% out of rank	P-value
Nitrite µmol/mL blood	10.37 (5.72-24.17)	7.18 (4.94-11.87)	9.15 (6.18-23.99) ^a	24.35 (12.05-38.45) ^b,c			0.0001^*
Nitrite µmol/mL urine	4.09 (1.05-7.84)	2.4 (0.89-4.85)	4.45 (0.64-9.52) ^a	5.72 (3.91-8.79) ^b			0.0001^*
% methemoglobin	1.94 (1.16-2.53)	1.33 (0.9-2)	1.56 (1.1-2.08) ^a	2.7 (2.5-2.9) ^b,c	< 1.5	80.71	0.0001^*
Glucose mg/dL	90.35 (79.6-108.15)	86.2 (75.2-100)	88.6 (79.8-101)	103.2 (84.2-132) ^b,c	70-110	30.94	0.0001^*
Cholesterol mg/dL	164.85 (133.5-208.2)	151 (131-178.5)	175.1 (133.5-208.6) ^a	179.85 (136.4-225.8) ^b,c	< 200	29.29	0.0012^*
HDL mg/dL	52.2 (40.4-71)	58.3 (41.5-73)	49.4 (38.5-68.6) ^a	51.1 (41-69.4)	30-75	20.15	0.0684
LDL mg/dL	92 (74.5-132)	82.5 (66.4-104)	98 (79-135) ^a	96 (80-144) ^b	< 130	40.95	0.001^*
Triglyceride mg/dL	104.4 (73.5-145.2)	101.8 (67.1-131.2)	90.1 (69.7-134)	127.7 (96-172.4) ^b	< 150	23.57	0.0001^*

Parameters	All women n=420	Low n=139	Medium n=171	High n=110	Reference rank	P-value
TSH µUI/mL	1.74 (1.24-2.67)	1.63 (1.18-2.42)	2.02 (1.3-3.01) ^a	1.6 (1.17-2.49) ^c	0.4-4	0.0126^*
T3T ng/dL	113 (98.1-132)	110 (95.4-128)	115 (99-140) ^a	113 (100-128)	82-179	0.1564
fT3 pg/mL	3.48 (3.07-4.05)	3.49 (3.17-3.92)	3.59 (3.07-4.63) ^a	3.31 (3.05-3.67) ^{b, c}	1-4	0.0003^*
T4T µg/dL	7.7 (6.8-8.65)	7.85 (7.06-8.97)	7.46 (6.55-8.62) ^a	7.5 (6.8-8.32) ^c	4.5-12.5	0.0186^*
fT4 ng/dL	1.07 (0.97-1.17)	1.08 (1-1.19)	1.03 (0.94-1.13) ^a	1.11 (0.99-1.2) ^c	0.3-6	0.0004^*
Subclinical hypothyroidism	35 (8.33)	10 (7.19)	19 (11.11)	6 (5.45)		0.206§

Genotype	All women n=420	Low n=139	Medium n=171	High n=110	P-value
FOXE1 rs965513
Genotype GG GA AA	241 (57.4%) 150 (35.7%) 29 (6.9%)	84 (60.5%) 47 (33.8%) 8 (5.7%)	107 (62.6%) 55 (32.2%) 9 (5.2%)	50 (45.5%) 48 (43.6%) 12 (10.9%)	0.042 0.43
Allele G A HWE	632 (75%) 208 (25%)	215 (77%) 63 (23%)	269 (78%) 73 (22%)	148 (67%) 72 (33%)
FOXE1 rs1867277
Genotype GG GA AA	214 (51%) 162 (38.6%) 44 (10.4%)	82 (59%) 45 (32.4%) 12 (8.6%)	99 (58%) 56 (32.7%) 16 (9.3%)	33 (30%) 61 (55.5%) 16 (14.5%)	0.0001 0.11
Allele G A HWE	590 (70%) 250 (30%)	209 (75%) 69 (25%)	254 (75%) 88 (25%)	127 (58%) 93 (42%)

Parameter	All women n=420	Low n=139	Medium n=171	High n=110	Reference rank	P-value
Index of nuclear division	1.85 (1.82-1.86)	1.84 (1.81-1.86)	1.85 (1.83-1.87)	1.85 (1.78-1.87)	1.3-2.2	0.0896
Mononuclear cells %	15.5 (12.7-18.2)	16.9 (13.9-19.4)	15.8 (12.9-17.9) ^a	13 (10.8-15.6) ^b,c	-	0.0001^*
Binucleated cells %	34.9 (29.7-39.4)	38.4 (34.6-40.1)	32.6 (27.3-38.4) ^a	34.2 (26.5-39.3) ^c	30-60	0.0001^*
Multinucleated cells	22 (19-26)	21 (18-26)	23 (19-26) ^a	21.5 (19-26)	-	0.0711
Apoptosis cells %	15 (11-19)	16 (11-21)	15 (12-19)	13 (11-19)	0-7	0.4990
Necrotic cells %	11 (8-16)	12 (9-16)	11 (8-16)	11 (8-18)	0-9	0.7126
Micronucleus %	38.9 (35.6-45.6)	35.2 (31.6-38.7)	41.8 (37.1-47.7) ^a	42.6 (37.6-48.3) ^c	0-30	0.0001^*
Cells with bridge %	24 (19-29)	24 (19-29)	24 (19-28)	25.5 (21-31)	0-10	0.0321
Cells with bubble %	25 (19-31)	25 (19-34)	23 (18-28) ^a	26 (21-34)	0-5	0.0021^*

	TSH µUI/mL			T4 ng/dL			T4 free			T3 pg/mL			T3 free
	β	SE	p-value	β	SE	p-value	β	SE	p-value	β	SE	p-value	β	SE	P-value
Low exposure	1			1			1			1			1
Medium exposure High exposure	0.21 -0.01	0.08 0.08	0.007^* 0.833	-0.07 -0.03	0.02 0.03	0.008^* 0.298	-0.03 -0.0007	0.02 0.02	0.070 0.972	0.07 0.02	0.03 0.03	0.008^* 0.447	0.12 -0.007	0.03 0.03	0.0001^* 0.822
Nitrite µmol/mL blood	0.02	0.03	0.500	0.002	0.01	0.843	0.006	0.008	0.433	0.001	0.01	0.882	-0.007	0.01	0.571
Nitrite µmol/mL urine	-0.03	0.02	0.180	-0.01	0.007	0.018^*	0.013	0.005	0.012^*	0.005	0.007	0.463	0.005	0.008	0.521
% <Methemoglobin	0.002	0.04	0.966	-0.005	0.02	0.716	-0.007	0.01	0.542	0.02	0.02	0.274	0.005	0.02	0.770
Glucose mg/dL	0.08	0.10	0.417	-0.04	0.03	0.217	-0.004	0.02	0.854	-0.09	0.03	0.007^*	-0.07	0.04	0.080
Cholesterol mg/dL	-0.05	0.09	0.549	-0.07	0.03	0.030^*	-0.04	0.02	0.092	-0.03	0.03	0.311	-0.05	0.03	0.125
HDL mg/dL	0.02	0.07	0.725	-0.005	0.02	0.851	0.006	0.02	0.761	0.03	0.03	0.264	0.008	0.03	0.788
LDL mg/dL	0.12	0.09	0.182	0.02	0.03	0.451	0.05	0.02	0.021^*	0.03	0.03	0.309	-0.011	0.03	0.463
Triglyceride mg/dL	0.06	0.06	0.317	0.002	0.02	0.893	0.008	0.01	0.581	0.07	0.02	0.0001^*	0.018	0.02	0.419
FOXE1 rs965513 GG GA AA	1 -0.17 -0.40	0.06 0.13	0.015^* 0.002^*	1 -0.03 -0.11	0.02 0.04	0.204 0.017^*	1 -0.02 -0.03	0.01 0.03	0.258 0.301	1 0.02 0.09	0.02 0.05	0.299 0.039^*	1 0.03 0.10	0.02 0.05	0.278 0.048^*
FOXE1 rs1867277 GG GA AA	1 -0.001 -0.09	0.07 0.11	0.984 0.418	1 0.002 0.01	0.02 0.04	0.904 0.751	1 0.02 0.03	0.02 0.03	0.100 0.312	1 0.02 0.03	0.02 0.04	0.241 0.418	1 0.006 -0.02	0.03 0.04	0.814 0.611
% Binucleated cells with MN	0.0002	0.0004	0.605	0.0001	0.0001	0.643	-0.0002	0.0001	0.034^*	0.0001	0.0001	0.754	-0.0001	0.0001	0.956

[1] Send correspondence to
Rebeca Pérez-Morales. Universidad Juárez del Estado de Durango, Facultad de Ciencias Químicas, Laboratorio de Biología Celular y Molecular, Av. Artículo, 123, Fracc., Filadelfia, C.P. 35010. Gómez Palacio, Durango, México. E-mail: rebecapms@ujed.mx

Brasil

Brasil

FOXE1 polymorphisms and chronic exposure to nitrates in drinking water cause metabolic dysfunction, thyroid abnormalities, and genotoxic damage in women

Abstract