Abstract
This study aimed to evaluate the effects of purple grape juice consumption in pregnancy on oxidative stress parameters in Wistar rat fetuses. Twenty-four pregnant rats were divided into five groups: control group, indomethacin group (received a single dose of indomethacin in DG20), group grape juice DG14 (received an amount for 14 days/first and second gestational trim), group grape juice DG20 (received a dose throughout the gestational period), group grape juice two doses (received two doses, at morning and afternoon). On the 20th day of pregnancy (DG20), rats were anesthetized, and a cesarean section was performed to obtain the fetuses. A sample of liver, heart, and total brain of fetuses was collected for oxidative stress analyses. Values P<0.05 were considered significant. In fetuses’ heart, we observed that the grape juice two dose group decreased sulfhydryl and increased SOD. In the liver, the grape juice decreased TBARS and SOD. There was a decrease in carbonyl and sulfhydryl in the indomethacin and grape juice one dose groups in the brain. We conclude that indomethacin altered oxidative stress parameters only in the fetal brain, and grape juice was presented as an important modulator of antioxidant capacity when consumed in a dose.
Keywords: Grape juice, offspring, pregnancy, indomethacin, oxidative stress
Introduction
Pregnancy is considered a moment of extreme importance in women’s life. In this period, physiological and nutritional changes occur, which can alter maternal metabolism [1]. Also, currently, there is an increase in exposure to free radicals and oxidative stress [2,3]. As a result, fetal circulation is different from the circulation of the newborn. At this stage, the placenta is responsible for the nutrition of the conceptus [4]. However, some organs such as the liver, heart, and brain of the fetus have already been functional since the beginning of development [5]. Therefore, both pre-gestational and gestational nutritional statuses are fundamental, as they are causally related to fetal growth and the development of specific pathologies of this period [6,7].
In this sense, it is worth emphasizing the importance of maintaining good eating habits [1], especially foods capable of acting in the body’s chemical reactions generating health benefits at different stages of life. These should receive special attention [8]. A balanced diet in macro and micronutrients is related to comorbidity prevention, avoiding the need for the use of contraindicated drugs during pregnancy [9], such as nonsteroidal anti-inflammatory drugs (NSAIDs) because these induce an imbalance in the synthesis of prostaglandins. Therefore, the number of studies related to NSAIDs and the consumption of polyphenols with the closure of the fetal arteriosus duct (DA) [10-12]. Furthermore, the vasoconstriction process activated by the use of NSAIDs during pregnancy is related to an increase in the production of reactive oxygen species (ROS) altering cell homeostasis [13], since the increased oxygen demand and energy substrate generation favor the process of oxidative stress during pregnancy [14]. However, studies are rare in the literature that demonstrates this interaction in foetal tissues.
However, several studies have analyzed the influence of maternal intake of foods with antioxidant and anti-inflammatory potential in offspring [15-18]. In this sense, the grape and its derivatives become a reference for its high concentration in phenolic compounds [19,20]. A recently published experimental study from our group observed a decrease in damage to proteins (carbonyls) in daughters’ hippocampus after maternal consumption of grape juice during pregnancy, demonstrating the benefits of these foods in transgenerational models [21].
In this sense, further studies about the polyphenols intake during pregnancy and its action on the offspring’s health are needed. Therefore, this study aimed to evaluate the effects of chronic consumption of purple grape juice at different periods and doses during pregnancy on oxidative stress parameters in the liver, heart, and total brain of Wistar rat fetuses.
Materials and methods
Animals
For this study, 22 female rats and 11 male rats of the species (Rattus norvegicus), Wistar, were heterogenic, approximately three months of age, from the bioterium of the Methodist University Center-Porto Alegre Institute. All animals were fed commercial feed and water at will and housed in plastic cages in the experimental rooms of the Bioterium itself, which have a temperature of 22±1°C, humidity between 60-80%, and light/dark cycle of 12 hours. It is noteworthy that males were used only for mating with females. All experimenters’ procedures were performed with the approval of the Ethics Committee on The Use of Animals (CEUA) of the Methodist University Center IPA, under protocol number 003/2017 and 001/2018.
Diet and grape juice
The palleted commercial food consisted of 20.5% protein (predominantly soybean), 54% carbohydrates, 4% fat, 4.5% fiber, 7% ash, and 10% moisture. The integral purple grape juice of the Vitis labrusca Bordeaux species was kindly donated by Nova Aliança. The juices were from the same harvest (2017) and the same batch and were previously analyzed. The total phenolic compounds (2.08±0.05), total flavonoids (2.58±0.002), catechin (15.41±0.59), epicatechin (3.08±0.06), epigallocatechin (0.33±0.005), epigallocatechin gallate (4.13±0.05) and gallic acid (4.92±0.11) were quantified.
Experimental design
The rats were placed for mating, in proportion to two females for one male per box in the late afternoon, and the vaginal smear was performed on the morning of the following day. Gestational day 0 (DG0) was determined by the presence of sperm in the vaginal smear, confirming pregnancy [22]. After the confirmation of pregnancy, pregnant rats were randomly divided into five experimental groups, as follows:
Group 1 (control), composed of five rats who received a daily morning administration of water at the dose of 7 μL/g of animal weight orally (gavage) from gestational day 0 (DG0) until the 20th day (DG20); Group 2 (indomethacin), composed of four rats who received a daily morning administration of water at the dose of 7 μL/g of animal weight orally (gavage) from DG0 and a single administration of indomethacin (Aspen Pharma, Durban, South Africa) at the dose of 10 mg/kg of animal weight orally (gavage) on the 20th day of pregnancy (DG20); Group 3 (DG14), composed of four rats who received a daily morning administration of integral purple grape juice at the dose of 7 μL/g of animal weight orally (gavage) from gestational day 0 (DG0) until the 14th day (DG14) characterizing first and second trimester; Group 4 (DG20), composed of four rats who received a daily morning administration of whole purple grape juice at the dose of 7 μL/g of animal weight orally (gavage) from gestational day 0 (DG0) until the 20th day (DG20); Group 5 (2 doses), composed of five rats who received two daily administrations of grape juice at the dose of 7 μL/g of animal weight each, one in the morning and one in the afternoon orally (gavage) from gestational day 0 (DG0) until the 20th day (DG20).
During treatment (pregnancy), the rats and their food ration were weighed daily on a digital scale (Crystal 200, Gibertini, Italy). Four hours after of administration of the last dose of gavage, the rats were anesthetized with an association of ketamine (75 mg/kg) and xylazine (10 mg/kg) administered intraperitoneally, and a cesarean section was performed to obtain the foetuses, which were euthanized before the first breath. After this procedure, the mothers were euthanized by cardiac puncture (exsanguination). Confirmation of euthanasia was performed by the evaluation of respiratory and cardiac arrest. At the end of this procedure, fetuses’ liver, heart, and total brain were extracted.
Oxidative stress parameters
Oxidative stress parameters were evaluated in the liver, heart, and total brain of foetuses. The tissues were homogenized in a 1,5% KCl solution and frozen (-20°C) until oxidative stress analysis.
Lipid peroxidation levels
The levels of thiobarbituric acid reactive substances (TBARS) were used to evaluate lipid peroxidation. The TBARS assay was performed according to the method described by Ohkawa et al. [23] Briefly, 50 µL of 8.1% sodium dodecyl sulfate (SDS), 375 µL of 20% acetic acid (pH 3.5), and 375 µL of 0.8% thiobarbituric acid (TBA-Merck, Darmstadt, Germany) were added to 200 µL of the sample and then incubated in a boiling water bath for 60 min. After cooling, the mixture was centrifuged (1000 g, 10 min). Next, the supernatant was removed, and absorbance was read at 535 nm on a spectrophotometer (T80 UV/VIS Spectrometer, PG Instruments, Alma Parck, Lutterworth, UK). Commercially available malondialdehyde was used as a standard. Results were expressed as nmol/mg protein.
Levels of oxidatively damaged proteins
The oxidation of serum proteins (carbonyls) was evaluated based on a reaction with 2,4-dinitrophenylhydrazine (DNPH-Sigma, St. Louis, USA) [24]. Briefly, a sample containing 1 mg of protein was reacted with 10 mM 2,4-dinitrophenylhydrazine for 30 min and subjected to protein precipitation with 10% TCA (Trichloroacetic Acid), followed by centrifugation (11,000 g, 3 min, 4°C). Next, the pellet was washed with ethanol:ethyl acetate (1:1) and centrifuged (11,000 g, 3 min, 4°C) three times. Then, pellets were suspended in 6 M guanidine hydrochloride (20 mM KH2PO4, pH 2.4) and read using a spectrophotometer (380 nm). Finally, blank samples were reacted with 2 M HCl instead of 10 mM 2,4-dinitrophenylhydrazine, and were run in parallel. Results were expressed as nmol carbonyl/mg protein.
Measurement of total sulphydryl groups
Quantifying the concentration of total sulfhydryl groups provides an idea of the level of oxidative attack on plasma proteins and is based on the reaction with 5,5’-Dithio-Bis (2-Nitrobenzoic Acid) [25]. Briefly, 50 μL of tissue or serum was mixed with 980 μL of PBS and 35 μL of 30 mM Tris/3 mM EDTA (pH 8) in a microplate well. After reading baseline absorbances, samples were reacted with 10 μL of 5,5’-dithiobis-(2-nitrobenzoic Acid) (10 mM in ethanol) for one hour. Absorbance was determined spectrophotometrically at 405 nm. The obtained values were compared with those obtained with a cysteine standard curve, and results were expressed as nmol-SH/mg protein.
Measurement of antioxidant enzyme activity
Using the method described by Aebi et al., catalase activity (CAT) was determined using the absorbance of hydrogen peroxide at 240 nm (25°C) [26]. The results were expressed as CAT/mg protein units. Superoxide dismutase (SOD) activity was determined by spectrophotometry based on a decrease in the autocatalytic adrenochrome formation rate at 480 nm [27]. Enzymatic activity was expressed as SOD/mg of protein units.
Protein dosage
Proteins were quantified using the Lowry et al. [28] method, using bovine serum albumin at the standard concentration of 1 mg/mL.
Statistical analysis
The values were determined as parametric or non-parametric, using the Kolmogorov-Sminorv. Non-parametric data were analyzed by Kruskal Wallis, followed by post-test Dunn’s, being expressed as the median and interquartile interval. Parametric data were analyzed by ANOVA from a pathway, followed by post-test Tukey, being expressed as the average and standard error. Values (P≤0.05) indicate a significant difference. All analyses were performed using sigma software Plot version 11.0.
Results
About the oxidative stress parameters, we observed that grape juice consumption during the pregnancy reduced the lipid peroxidation in the liver of fetuses (P≤0.05) (Figure 1A). In oxidative protein damage, we observed that grape juice two doses increased compared to the control and indomethacin groups (P≤0.05) (Figure 1B). Concerning antioxidant enzymes, we observed that the intake of grape juice in the first trimesters of pregnancy or two doses decreased enzyme activity superoxide dismutase compared to the indomethacin group (P≤0.05) (Figure 1D). However, there were no significant differences in sulfhydryl levels and catalase activity (Figure 1C and 1E).
Figure 1.
Analysis of cellular damage parameters and antioxidant capacity. Lipid peroxidation levels (TBARS) (A). Levels of protein oxidation (carbonyls) (B). Levels of sulfhyidryls grouping (C). Levels of activity of the enzyme superoxide dismutase (SOD) (D). Levels of catalase enzyme activity (CAT) (E) in the liver of Wistar rat fetuses that were treated with purple grape juice during the gestational period. Data expressed in median and interquartile interval. *Difference between these groups in relation to the control group. #Difference between the groups in relation to the indomethacin group. Statistical difference according to ANOVA of a pathway, with post Dunn’s test, P<0.05.
Regarding the analysis of these parameters in the heart of foetuses we observed that grape juice and indomethacin could not alter lipid damage markers and proteins (Figure 2A and 2B). However, the intake of 2 doses of grape juice decreases sulfhydryl amount. It increases superoxide dismutase activity compared to the one dose grape juice group (P≤0.05) (Figure 2C and 2D), no significant difference was observed in the catalase enzyme (Figure 2E).
Figure 2.
Analysis of cellular damage parameters and antioxidant capacity. Lipid peroxidation levels (TBARS) (A). Levels of protein oxidation (carbonyls) (B). Levels of sulfhyidryls grouping (C). Levels of activity of the enzyme superoxide dismutase (SOD) (D). Levels of catalase enzyme activity (CAT) (E) in the heart of Wistar rat fetuses that were treated with purple grape juice during the gestational period. Data expressed on average and standard error. $Difference between these groups grape juice DG20 and grape juice 2 doses. Statistical difference according to ANOVA of a pathway, with Tukey’s post-test, P<0.05.
Regarding the brain tissue of fetuses, we observed that the grape juice two doses decreased TBARS levels (P≤0.05) (Figure 3A). Indomethacin and grape juice one dose decreased compared to the control groups, DG14 grape juice, and group 2 doses for carbonyl levels (P≤0.05) (Figure 3B). About the sulfhydryl group, indomethacin group, and grape juice, one dose showed a decrease compared to the control groups, DG14 grape juice, and group 2 doses (P≤0.05) (Figure 3C). However, both juice and indomethacin did not alter SOD and CAT activity (Figure 3D and 3E).
Figure 3.
Analysis of cellular damage parameters and antioxidant capacity. Lipid peroxidation levels (TBARS) (A). Levels of protein oxidation (carbonyls) (B). Levels of sulfhyidryls grouping (C). Levels of activity of the enzyme superoxide dismutase (SOD) (D). Levels of catalase enzyme activity (CAT) (E) in the total brain of Wistar rat fetuses that were treated with purple grape juice during the gestational period. Data expressed in median and interquartile interval and carbonyls expressed on average and standard error. *Difference between these groups in relation to the control group. #Difference between the groups in relation to the group indomethacin. $Difference between the grape juice groups. Statistical difference according to ANOVA of a pathway, with post test of Dunn’s or Tukey, P<0.05.
Discussion
Studies by our research group have demonstrated the importance and benefits of consuming grape juice during pregnancy [18,21]. Given this, our study evaluated the consumption of grape juice at different stages and doses during the gestational period, evaluating fetuses’ liver, heart, and brain tissue. This study investigated the relationship between maternal food consumption and the impacts caused on the offspring compared with indomethacin (an anti-inflammatory drug) during the gestational period. As a result, we can see that grape juice decreased lipid peroxidation in the liver, increased carbonyl and decreased superoxide dismutase, increased sulfhydryl levels in the heart. In addition to reducing lipid peroxidation, protein damage and altering sulfhydryl levels in brain tissue, and demonstrating that maternal nutrition influences the redox homeostasis of the offspring [18,21,29].
By evaluating, in detail, the biomarkers of oxidative stress in the liver of fetuses, we observed a decrease in lipid peroxidation in the groups whose mothers ingested grape juice. This result corroborates Vega et al. [30], who observed protection from lipid peroxidation in fetuses of mothers who received resveratrol and a low-protein diet during pregnancy. This protection demonstrated that grape compounds could protect against the damage caused by an inadequate diet. Also, in the liver, we observed that grape juice 2 doses, ingested in excess, increased protein oxidation compared to the control group. In a recent experimental study, ethanol exposed during pregnancy increased protein oxidation levels compared to control, demonstrating that the fetal liver is an organ affected by maternal ingestion [31]. Thus, possibly justifying that the excess doses of foods even rich in polyphenols should be controlled during pregnancy, as they can alter the redox homeostasis in fetal liver tissue. However, the literature already described that the consumption of grape juice during the gestational period decreased the levels of carbonyl in the mothers’ liver. However, it could not protect against the damage caused by the ingestion of a diet rich in lipids [29].
Regarding the antioxidant defenses in the liver of fetuses, we observed a decrease in the superoxide dismutase enzyme activity in the groups that consumed grape juice compared to the indomethacin group. Corroborating our study, Vega et al. [30] also observed a decrease in SOD in the liver of fetuses of mothers who ingested a low-protein diet with resveratrol administration compared to the control group. One explanation for this occurrence is that antioxidants play the role of the enzyme, acting as “SOD-like”, thus generating a decrease in enzyme activity [32].
Our study observed changes in the antioxidant defense system in the heart of fetuses generated by the consumption of grape juice in two doses, with a decrease in the content of sulfhydryls and an increase in SOD compared to the grape juice group DG20. Furthermore, Tarry-adkins et al. [33], evaluated the influence of a low-protein diet during pregnancy followed by a recovery from low birth weight through breastfeeding. After 3 and 22 days of birth, the authors observed an increase in catalase expression and CuZnSOD decrease in the hearts of puppies with weight recovery. The results were more significant at 22 days after birth, positively reflecting maternal nutrition’s influence on the antioxidant capacity. But it is important to mention that the adequate amount of grape juice ingested during pregnancy did not influence cell damage markers such as TBARS and Carbonyl. Also, it didn’t alter the activity of antioxidant enzymes in fetuses’ heart, as there were no differences in the control group.
Indomethacin is an NSAID widely used in AD constriction induction models [34-36]. Still, the counterpoint, although contraindicated, is a drug widely used during pregnancy to combat fever, pain, and inflammation due to a lack of clarification of its adverse effects [37]. However, the results of this drug on the fetus are rare in the literature since the main organ involved in the metabolization of this drug is the maternal liver, and there is a passage, even in a short period, to the fetus through the placenta [38,39]. In our study, we observed that indomethacin did not damage lipids and proteins, nor did it alter the antioxidant activity in the liver and heart of fetuses. Perhaps the explanation is the dose and time of action since the drug was administered four hours before the cesarean. However, it showed a relative change in the fetal brain.
Brain tissue is rich in polyunsaturated fatty acids, making it highly susceptible to free radicals [40]. Our study observed that grape juice in 2 doses decreased lipid peroxidation in the fetal brain compared to the control group. Previous studies by our group have already highlighted the beneficial effects of grape juice on brain tissue, corroborating our findings [20,41,42]. However, regarding protein oxidation, we observed that both indomethacin and DG20 grape juice decreased levels in brain tissue compared to the control group. The effects of indomethacin on protein oxidation in the brain have not been described in the literature. However, indomethacin increased carbonyl levels in gastric tissue in an experimental study [43]. As for the decrease in the oxidation of proteins generated by grape juice, Gabardo et al. [44], evaluated the effects of grape juice on brain tissue and observed a decrease in carbonyl levels in the hippocampus after in vitro incubation of grape juice and also protection of grape juice against increased damage from protein oxidation generated by temozolomide (TMZ) in the cortex, hippocampus and cerebellum.
Regarding the analysis of the antioxidant capacity in the fetal brain, we only observed changes in the levels of the sulfhydryl group, where indomethacin decreased the levels of non-enzymatic activity. Hilal et al. [45] evaluated the effect of indomethacin at different concentrations in brain tissue of male rats, and observed a significant decrease in glutathione reductase (GSH), as well as in the activity of SOD and CAT, demonstrating the genotoxic effects of this drug’s anti-inflammatory drugs. However, unlike this study, we did not observe changes in the activity of superoxide dismutase and catalase enzymes. In our research, grape juice also decreased non-enzymatic activity levels. This result differs from another study from our group that demonstrated that grape juice consumption during pregnancy did not change sulfhydryl levels in rats’ cortex, hippocampus, and cerebellum [42]. However, grape juice associated with gavage stress generated an increase in sulfhydryl in the cerebral cortex, demonstrating that grape juice and other important factors from this period contribute to the alteration of oxidative stress biomarkers [42].
Assessing the results obtained in our study, we can conclude that indomethacin administered at the end of pregnancy changed oxidative stress parameters only in fetal brain tissue. Moreover, it was realized that the consumption of grape juice during pregnancy could reduce or modulate oxidative damage and antioxidant defenses in fetuses. Thus, we can define that the daily and moderate consumption of grape juice promotes better results when compared to the data obtained in the groups that consumed only 14 days of gestation or ingested the drink in two doses. Despite this, further studies should be encouraged, including knowing the effects of using anti-inflammatory drugs for a more extended period and their action on the offspring. Despite this, we can say that the consumption of grape juice by pregnant women helps balance the antioxidant defense in their offspring.
Acknowledgements
The authors would like to thank the Methodist University Center-IPA, Ibravin (the Brazilian Wine Institute) provided the grape juice to the experiment. CAPES (Coordination for the Improvement of Higher Education Personnel), FAPERGS (Research Support Foundation of the State of Rio Grande do Sul), CNPq (National Council for Scientific and Technological Development).
Disclosure of conflict of interest
None.
References
- 1.Baião MR, Deslandes SF. Alimentação na gestação e puerpério. Rev Nutr. 2006;19:245–53. [Google Scholar]
- 2.Agarwal A, Gupta S, Sikka S. The role of free radicals and antioxidants in reproduction. Curr Opin Obstet Gynecol. 2006;18:325–32. doi: 10.1097/01.gco.0000193003.58158.4e. [DOI] [PubMed] [Google Scholar]
- 3.Vannucchi CI, Jordao AA, Vannucchi H. Antioxidant compounds and oxidative stress in female dogs during pregnancy. Res Vet Sci. 2007;83:188–93. doi: 10.1016/j.rvsc.2006.12.009. [DOI] [PubMed] [Google Scholar]
- 4.Finnemore A, Groves A. Physiology of the fetal and transitional circulation. Semin Fetal Neonatal Med. 2015;20:210–6. doi: 10.1016/j.siny.2015.04.003. [DOI] [PubMed] [Google Scholar]
- 5.Rudolph AM. Hepatic and ductus venosus blood flows during fetal life. Hepatology. 1983;3:254–8. doi: 10.1002/hep.1840030220. [DOI] [PubMed] [Google Scholar]
- 6.Bonfim C. Nutritional status and gestational complications: a review. Rev Saúde Com. 2014;10:409–21. [Google Scholar]
- 7.August PM, Maurmann RM, Saccomori AB, Scortegagna MC, Flores EB, Klein CP, Santos BG, Stone V, Dal Magro BM, Cristhian L, Santo CN, Hözer R, Matté C. Effect of maternal antioxidant supplementation and/or exercise practice during pregnancy on postnatal overnutrition induced by litter size reduction: brain redox homeostasis at weaning. Int J Dev Neurosci. 2018;71:146–55. doi: 10.1016/j.ijdevneu.2018.09.003. [DOI] [PubMed] [Google Scholar]
- 8.Gernand AD, Schulze KJ, Stewart CP, West KP Jr, Christian P. Micronutrient deficiencies in pregnancy worldwide: health effects and prevention. Nat Rev Endocrinol. 2016;12:274–89. doi: 10.1038/nrendo.2016.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Ferraz L, Albiero C, Boechat SG, Fonseca IP, Farias VP, Braga A, Lopes PF. Micronutrients and their importance in the gestational period. Rev Sabber Scientific. 2018;7:68–82. [Google Scholar]
- 10.Fonseca CS, Viloria MIV, Repetti L. Fetal changes induced by the use of anti-inflammatory drugs during pregnancy. Rural Science. 2002;32:529–34. [Google Scholar]
- 11.Zielinsky P, Piccoli AL Jr, Manica JL, Nicoloso LH. New insights on fetal ductal constriction: role of maternal ingestion of polyphenol-rich foods. Expert Rev Cardiovasc Ther. 2010;8:291–8. doi: 10.1586/erc.09.174. [DOI] [PubMed] [Google Scholar]
- 12.Zielinsky P, Manica JL, Piccoli AL Jr, Nicoloso LH, Barra M, Alievi MM, Vian I, Zilio A, Pizzato PE, Silva JS, Bender LP, Pizzato M, Menezes HS, Garcia SC. Fetal ductal constriction caused by maternal ingestion of green tea in late pregnancy: an experimental study. Prenat Diagn. 2012;32:921–6. doi: 10.1002/pd.3933. [DOI] [PubMed] [Google Scholar]
- 13.Ovalı F. Molecular and mechanical mechanisms regulating ductus arteriosus closure in preterm infants. Front Pediatr. 2020;8:516. doi: 10.3389/fped.2020.00516. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Casanueva E, Viteri FE. Iron and oxidative stress in pregnancy. J Nutr. 2003;133(Suppl 2):1700S–1708S. doi: 10.1093/jn/133.5.1700S. [DOI] [PubMed] [Google Scholar]
- 15.Emiliano AF, de Cavalho LC, da Silva Cristino Cordeiro V, da Costa CA, de Oliveira PB, Queiroz EF, Col Moreira DD, Boaventura GT, de Moura RS, Resende AC. Metabolic disorders and oxidative stress programming in offspring of rats fed a high-fat diet during lactation: effects of a vinifera grape skin (ACH09) extract. J Cardiovasc Pharmacol. 2011;58:319–28. doi: 10.1097/FJC.0b013e3182244a51. [DOI] [PubMed] [Google Scholar]
- 16.Arola-Arnal A, Oms-Oliu G, Crescenti A, del Bas JM, Ras MR, Arola L, Caimari A. Distribution of grape seed flavanols and their metabolites in pregnant rats and their fetuses. Mol Nutr Food Res. 2013;57:1741–52. doi: 10.1002/mnfr.201300032. [DOI] [PubMed] [Google Scholar]
- 17.Hachul ACL, Boldarine VT, Neto NIP, Moreno MF, Carvalho PO, Sawaya ACHF, Ribeiro EB, Oller do Nascimento CM, Oyama LM. Effect of the consumption of green tea extract during pregnancy and lactation on metabolism of mothers and 28d-old offspring. Sci Rep. 2018;8:1869. doi: 10.1038/s41598-018-20174-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18.Hilger DK, Wohlenberg MF, Schaffer TK, Machado FS, Gonçalves LK, Bortolato G, Dani G, Rodrigues A, Funchal C, Dani C. Purple grape juice, an important flavonoids source, influence in biochemical parameters in offspring of Wistar rats. Food and Nutr Sci. 2015;6:683–91. [Google Scholar]
- 19.Abe LT, Mota RV, Lajolo FM, Genovese MI. Phenolic compounds and antioxidant capacity of cultivars of Grapes Vitis labrusca L. and Vitis vinifera L. Ciênc Tecnol Aliment. 2007;27:394–400. [Google Scholar]
- 20.Dani C, Oliboni LS, Agostini F, Funchal C, Serafini L, Henriques JA, Salvador M. Phenolic content of grapevine leaves (Vitis labrusca var. Bordo) and its neuroprotective effect against peroxide damage. Toxicol In Vitro. 2010;24:148–53. doi: 10.1016/j.tiv.2009.08.006. [DOI] [PubMed] [Google Scholar]
- 21.Proença ICT, Gonçalves LK, Schmitz F, Mello A, Funchal CS, Wyse A, Dani C. Purple grape juice consumption during the gestation reduces acetylcholinesterase activity and oxidative stress levels provoked by high-fat diet in hippocampus from adult female rats descendants. An Acad Bras Cienc. 2021;93:e20191002. doi: 10.1590/0001-3765202120191002. [DOI] [PubMed] [Google Scholar]
- 22.Dias R, Trindade JCS, Rudge MVC. Experimental arterial hypertension and pregnancy in rats: repercussions on weight, length and organs of newborns. Rev Bras Ginecol and Obs. 2000;22:567–72. [Google Scholar]
- 23.Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95:351–8. doi: 10.1016/0003-2697(79)90738-3. [DOI] [PubMed] [Google Scholar]
- 24.Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER. Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 1990;186:464–78. doi: 10.1016/0076-6879(90)86141-h. [DOI] [PubMed] [Google Scholar]
- 25.Aksenov MY, Markesbery WR. Changes in thiol content and expression of glutathione redox system genes in the hippocampus and cerebellum in Alzheimer’s disease. Neurosci Lett. 2001;302:141–145. doi: 10.1016/s0304-3940(01)01636-6. [DOI] [PubMed] [Google Scholar]
- 26.Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121–6. doi: 10.1016/s0076-6879(84)05016-3. [DOI] [PubMed] [Google Scholar]
- 27.Bannister JV, Calabrese L. Assays for superoxide dismutase. Methods Biochem Anal. 1987;32:279–312. doi: 10.1002/9780470110539.ch5. [DOI] [PubMed] [Google Scholar]
- 28.Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75. [PubMed] [Google Scholar]
- 29.Gonçalves L, Bortolato G, Braccini Neto R, Frusciante MR, Funchal C, Dani C. Grape juice consumption with or without high fat diet during pregnancy reduced the weight gain and improved lipid profile and oxidative stress levels in liver and serum from Wistar rats. Beverages. 2018;4:78. [Google Scholar]
- 30.Vega CC, Reyes-Castro LA, Rodríguez-González GL, Bautista CJ, Vázquez-Martínez M, Larrea F, Chamorro-Cevallos GA, Nathanielsz PW, Zambrano E. Resveratrol partially prevents oxidative stress and metabolic dysfunction in pregnant rats fed a low protein diet and their offspring. J Physiol. 2016;594:1483–99. doi: 10.1113/JP271543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31.Ince E, Curabeyoğlu F, Akyol S. Oxidative stress in lymphoid tissues and complement activation in alcoholic mother rats and their newborns. Gen Physiol Biophys. 2019;38:91–100. doi: 10.4149/gpb_2018018. [DOI] [PubMed] [Google Scholar]
- 32.Schaffer TK, Wohlenberg MF, Medeiros N, Martins JB, Agostini F, Funchal C, Dani C. Evaluation of antioxidant activity of grapevine leaves extracts (Vitis labrusca) in liver of Wistar rats. An Acad Bras Cienc. 2016;88:187–96. doi: 10.1590/0001-3765201620140658. [DOI] [PubMed] [Google Scholar]
- 33.Tarry-Adkins JL, Martin-Gronert MS, Fernandez-Twinn DS, Hargreaves I, Alfaradhi MZ, Land JM, Aiken CE, Ozanne SE. Poor maternal nutrition followed by accelerated postnatal growth leads to alterations in DNA damage and repair, oxidative and nitrosative stress, and oxidative defense capacity in rat heart. FASEB J. 2013;27:379–90. doi: 10.1096/fj.12-218685. [DOI] [PubMed] [Google Scholar]
- 34.Takami T, Momma K, Imamura S. Increased constriction of the ductus arteriosus by dexamethasone, indomethacin, and rofecoxib in fetal rats. Circ J. 2005;69:354–8. doi: 10.1253/circj.69.354. [DOI] [PubMed] [Google Scholar]
- 35.Beaute JI, Friedman KG. Indomethacin induced ductus arteriosus closure in midgestation fetus. Clin Case Rep. 2018;6:506–8. doi: 10.1002/ccr3.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 36.Erenel H, Karslı MF, Korkmaz SÖ, Şen C. Are complicated monochorionic twins more susceptible to indomethacin-induced fetal ductal constriction? Two cases of laser surgery for twin-twin transfusion syndrome. Turk J Obstet Gynecol. 2018;15:200–3. doi: 10.4274/tjod.78095. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 37.Antonucci R, Zaffanello M, Puxeddu E, Porcella A, Cuzzolin L, Pilloni MD, Fanos V. Use of non-steroidal anti-inflammatory drugs in pregnancy: impact on the fetus and newborn. Curr Drug Metab. 2012;13:474–90. doi: 10.2174/138920012800166607. [DOI] [PubMed] [Google Scholar]
- 38.Morozova EV. Structure of the mesenteric lymph nodes in fetuses and offspring of white rats under the effect of indomethacin on the mother-fetus system. Arkh Anat Gistol Embriol. 1989;96:48–55. [PubMed] [Google Scholar]
- 39.Morozova EV, Moiseeva ML. Morphological characteristics of the thymus gland of albino rats after prenatal administration of indomethacin. Arkh Anat Gistol Embriol. 1990;98:33–8. [PubMed] [Google Scholar]
- 40.Bassett CN, Montine TJ. Lipoproteins and lipid peroxidation in Alzheimer’s disease. J Nutr Health Aging. 2003;7:24–9. [PubMed] [Google Scholar]
- 41.Rodrigues AD, Scheffel TB, Scola G, Santos MT, Fank B, de Freitas SC, Dani C, Vanderlinde R, Henriques JA, Coitinho AS, Salvador M. Neuroprotective and anticonvulsant effects of organic and conventional purple grape juices on seizures in Wistar rats induced by pentylenetetrazole. Neurochem Int. 2012;60:799–805. doi: 10.1016/j.neuint.2012.01.009. [DOI] [PubMed] [Google Scholar]
- 42.Wohlenberg M, Machado F, Shaffer T, Botolato G, Marinho JP, Medeiros NS, Mello A, Agostini F, Gerson S, Funchal C, Dani C. Effect of chronic treatment with grape juice during pregnancy and nursing in central nervous system of female Wistar rats. J Nutr Heal Food Eng. 2017;6:75–81. [Google Scholar]
- 43.Pineda-Peña EA, Martínez-Pérez Y, Galicia-Moreno M, Navarrete A, Segovia J, Muriel P, Favari L, Castañeda-Hemández G, Cháves-Pinã AE. Participation of the anti-inflammatory and antioxidative activity of docosahexaenoic acid on indomethacin-induced gastric injury model. Eur J Pharmacol. 2018;818:585–92. doi: 10.1016/j.ejphar.2017.11.015. [DOI] [PubMed] [Google Scholar]
- 44.Gabardo T, Frusciante R, Marinho JP, Santos M, Abujamra AL, Dani C, Funchal C. In vitro antioxidant activity of purple grape juice (vitis labrusca) against temozolamide treatment in rat brains. Beverages. 2018;4:99. [Google Scholar]
- 45.Hilal Ahmad M, Fatima M, Hossain MM, Mondal AC. Determination of potential oxidative damage, hepatotoxicity, and cytogenotoxicity in male Wistar rats: role of indomethacin. J Biochem Mol Toxicol. 2018;32:e22226. doi: 10.1002/jbt.22226. [DOI] [PubMed] [Google Scholar]