Prolonged use of finasteride-induced gonadal sex steroids alterations, DNA damage and menstrual bleeding in women

Abstract The aim of the present study was to examine the effect of prolonged use of finasteride on serum levels of dihydrotestosterone (DHT), estradiol (E2), progesterone, testosterone and androstenedione in women during the menstrual period. Further, to screen and compare the 5α-reductase activities through the expression of SRD5A1, SRD5A2 and AR gene and to determine the level of VEGF, VKOR and SAA gene expression and DNA damage. A total of 30 Saudi women aged between 25 and 35 years were enrolled in the study. The selected women were divided into two groups. The first group (n = 15) received 5 mg finasteride/day for prolonged period of one year and second group (n = 15) was taken as a healthy control. ELISA technique was used for measuring the serum levels of the targeted hormones, and Comet assay was used for checking the DNA integrity. Our findings revealed significant decrement of DHT, E2, progesterone and androstenedione levels and elevated levels of testosterone in group treated with daily oral doses of 5 mg finasteride/day compared with the control subjects. mRNA expression suggested that finasteride has concrete effects on the gene expression of the selected genes from the treated group in comparison with the control group. In addition, finasteride induced DNA damage, and heavy menstrual bleeding was noted in women treated with finasteride. In conclusion, the present findings revealed that finasteride has adverse health effects in women associated with gonadal sex steroids alterations, DNA damage and heavy menstrual bleeding with no consensus in the treatment of androgenetic alopecia in women.

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Anti-Obesity and Anti-Adipogenic Effects of Chitosan Oligosaccharide (GO2KA1) in SD Rats and in 3T3-L1 Preadipocytes Models

Molecules ◽

10.3390/molecules26020331 ◽

2021 ◽

Vol 26 (2) ◽

pp. 331

Author(s):

Jung-Yun Lee ◽

Tae Yang Kim ◽

Hanna Kang ◽

Jungbae Oh ◽

Joo Woong Park ◽

...

Keyword(s):

Gene Expression ◽

Body Weight ◽

Density Lipoprotein ◽

Treated Group ◽

Control Group ◽

Chitosan Oligosaccharide ◽

Sd Rats ◽

Adipogenic Gene

Excess body weight is a major risk factor for type 2 diabetes (T2D) and associated metabolic complications, and weight loss has been shown to improve glycemic control and decrease morbidity and mortality in T2D patients. Weight-loss strategies using dietary interventions produce a significant decrease in diabetes-related metabolic disturbance. We have previously reported that the supplementation of low molecular chitosan oligosaccharide (GO2KA1) significantly inhibited blood glucose levels in both animals and humans. However, the effect of GO2KA1 on obesity still remains unclear. The aim of the study was to evaluate the anti-obesity effect of GO2KA1 on lipid accumulation and adipogenic gene expression using 3T3-L1 adipocytes in vitro and plasma lipid profiles using a Sprague-Dawley (SD) rat model. Murine 3T3-L1 preadipocytes were stimulated to differentiate under the adipogenic stimulation in the presence and absence of varying concentrations of GO2KA1. Adipocyte differentiation was confirmed by Oil Red O staining of lipids and the expression of adipogenic gene expression. Compared to control group, the cells treated with GO2KA1 significantly decreased in intracellular lipid accumulation with concomitant decreases in the expression of key transcription factors, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (CEBP/α). Consistently, the mRNA expression of downstream adipogenic target genes such as fatty acid binding protein 4 (FABP4), fatty acid synthase (FAS), were significantly lower in the GO2KA1-treated group than in the control group. In vivo, male SD rats were fed a high fat diet (HFD) for 6 weeks to induced obesity, followed by oral administration of GO2KA1 at 0.1 g/kg/body weight or vehicle control in HFD. We assessed body weight, food intake, plasma lipids, levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) for liver function, and serum level of adiponectin, a marker for obesity-mediated metabolic syndrome. Compared to control group GO2KA1 significantly suppressed body weight gain (185.8 ± 8.8 g vs. 211.6 ± 20.1 g, p < 0.05) with no significant difference in food intake. The serum total cholesterol, triglyceride, and low-density lipoprotein (LDL) levels were significantly lower in the GO2KA1-treated group than in the control group, whereas the high-density lipoprotein (HDL) level was higher in the GO2KA1 group. The GO2KA1-treated group also showed a significant reduction in ALT and AST levels compared to the control. Moreover, serum adiponectin levels were significantly 1.5-folder higher than the control group. These in vivo and in vitro findings suggest that dietary supplementation of GO2KA1 may prevent diet-induced weight gain and the anti-obesity effect is mediated in part by inhibiting adipogenesis and increasing adiponectin level.

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Diabetes mellitus increased integrins gene expression in rat endometrium at the time of embryo implantation

International Journal of Reproductive BioMedicine ◽

10.18502/ijrm.v17i6.4810 ◽

2019 ◽

Author(s):

Abbas Bakhteyari ◽

Yasaman Zarrin ◽

Parvaneh Nikpour ◽

Zeinab Sadat Hosseiny ◽

...

Keyword(s):

Gene Expression ◽

Diabetes Mellitus ◽

Group Treatment ◽

Embryo Implantation ◽

Treated Group ◽

Diabetic Rats ◽

Diabetic Rat ◽

Control Group ◽

Genes Expression ◽

Normal Menstrual Cycle

Background: Diabetes mellitus deeply changes the genes expression of integrin (Itg) subunits in several cells and tissues such as monocytes, arterial endothelium, kidney glomerular cells, retina. Furthermore, hyperglycemia could impress and reduce the rate of successful assisted as well as non-assisted pregnancy. Endometrium undergoes thorough changes in normal menstrual cycle and the question is: What happens in the endometrium under diabetic condition? Objective: The aim of the current study was to investigate the endometrial gene expression of α3, α4, αv, Itg β1 and β3 subunits in diabetic rat models at the time of embryo implantation. Materials and Methods: Twenty-eight rats were randomly divided into 4 groups: control group, diabetic group, pioglitazone-treated group, and metformin-treated group. Real-time PCR was performed to determine changes in the expression of Itg α3, α4, αv, β1, and β3 genes in rat’s endometrium. Results: The expression of all Itg subunits increased significantly in diabetic rats’ endometrium compared with control group. Treatment with pioglitazone significantly reduced the level of Itg subunits gene expression compared with diabetic rats. While metformin had a different effect on α3 and α4 and elevated these two subunits gene expression. Conclusion: Diabetes mellitus significantly increased the expression of studied Itg subunits, therefore untreated diabetes could be potentially assumed as one of the preliminary elements in embryo implantation failure.

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69 AN EASY-TO-PERFORM METHOD TO ASSESS VIABILITY OF FELINE FREEZE-DRIED SPERM

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab69 ◽

2013 ◽

Vol 25 (1) ◽

pp. 182 ◽

Cited By ~ 3

Author(s):

L. C. O. Magalhães ◽

C. M. Melo-Oña ◽

M. J. Sudano ◽

D. M. Paschoal ◽

L. F. Crocomo ◽

...

Keyword(s):

Dna Damage ◽

Acridine Orange ◽

Freeze Drying ◽

Membrane Damage ◽

Daily Basis ◽

Control Group ◽

Dna Integrity ◽

Sperm Cells ◽

Significance Level ◽

Freeze Dried

Freeze-drying sperm seems to be the ultimate alternative to preserve sperm cells from endangered species because of its easiness in transportation and storage of samples. This technique has already been used for equine, bovine, murine, rabbit, canine, and feline sperm cells. However, it is still important to verify the DNA integrity of such samples to produce viable offspring. The objective of this study was to verify the reliability of acridine orange (AO) staining to assess DNA damage of feline freeze-dried sperm samples. Three normospermic cats were used as sperm donors, and sperm samples were collected with the use of an artificial vagina without the presence of an oestrous queen. The control group (G1) used only fresh semen. To increase the range of sperm variability before freeze-drying, the sperm samples were supplemented with 2 extenders, namely SOF (G2) and human tubal fluid (G3). These extenders were selected because of their lack of cryoprotectant agents with a view to producing membrane damage and thus putting DNA integrity at risk. Samples were taken to a freeze-dryer (Edwards do Brasil, Brazil) to produce stable free-dried sperm. Over 50 samples were assessed for DNA damage by using AO and alkaline comet assay (CA). Acridine orange taints red/orange the sperm cells that present sperm damage, and CA shows long comet tails for cells with DNA damage. The CA was selected to countercheck the AO results owing to its higher accuracy. Nevertheless, both AO and CA count 100 cells per analysis, and a total of 3 repetitions per sample were sufficient. For the statistical analysis, SAS PROC GLM was used, and the significance level was set at 0.05. The AO staining showed that the G1 samples had a greater DNA damage (27.1%) if compared with G2 (15.2%) and G3 (23.1%; P < 0.0001). Nevertheless, G2 and G3 displayed sufficient DNA integrity throughout the whole lyophilization process. The CA, which is a more precise evaluation technique, proved that the groups had distinguished results but still were ordered in the same way: G1 was the one with more DNA damage (17.7%), G3 was second (14.4%), and G2 was third (3.2%) in DNA alterations. Just like AO, the CA showed difference among treatments (P < 0.0001). Since AO is a technique that requires very little sophistication and presented about the same results as the very elaborate CA method, it might be used on a daily basis to assess feline sperm DNA that underwent the freeze-drying process.

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A Comparison of Thioredoxin 1 Gene Expression in Schizophrenia Patients before and after Treatment with Risperidone

10.53350/pjmhs211551551 ◽

2021 ◽

Vol 15 (5) ◽

pp. 1551-1563

Author(s):

E. Azizi ◽

M. Hosseinzadeh ◽

P. Vahdatian ◽

A. Adibi ◽

A. Azizifar ◽

...

Keyword(s):

Gene Expression ◽

Rna Extraction ◽

Serum Levels ◽

Key Words Schizophrenia ◽

First Episode ◽

Control Group ◽

Blood Leukocytes ◽

Before And After ◽

Schizophrenic Patients ◽

Dsm Iv

Background: Schizophrenia is a mental disorder characterized by distortions in thinking, perception, emotions, language, self-sense, and behavior. Recent research suggests that Reactive Oxygen Species (ROS) are involved in the pathophysiology of schizophrenia. Studies have also shown the increased plasma and serum levels of the Trx1 molecule in schizophrenia patients. In the present study, the researchers compared the expression levels of Trx1 mRNA in peripheral blood leukocytes of Iranian schizophrenia patients compared to healthy controls. Methods: First-episode patients (n=35) who met DSM-IV criteria for schizophrenia were recruited from patients referred to psychiatrists in the city of Ilam and Farabi Hospital in Kermanshah. Healthy people were also selected by recruiting people who, according to a psychiatrist, did not have any mental illness. Diagnoses were made for each patient by two independent experienced psychiatrists and confirmed by the Structured Clinical Interview for DSM-IV (SCID). Patients were treated with risperidone for three months and then compared with thirty- five healthy volunteers. Patients were sampled before and after treatment and then by RNA Extraction and DNA synthesis, Trx1 gene expression was performed by real-time PCR method. Results: Comparison of Trx1 gene expression in PBMCs of schizophrenic patients before and after treatment with the control group showed that the expression of Trx1 gene of the “before” treatment group was significantly increased compared with that of the control group (P= 0.0007). Also, Trx1 gene expression in PBMCs of “before” and “after “groups showed that Trx1 gene expression of “after” group was significantly decreased compared to the “before” group (P= 0.014). These results showed that the mean of positive, negative, and general psychopathology was reduced significantly in schizophrenic patients before and after treatment in all three cases (P <0.001). Conclusion: the expression of TRX in PBMCs of schizophrenic patients decreased after risperidone treatment. This reduction of expression was statistically significant and indicates the possible effect of risperidone on the expression of the TRX gene in PBMCs of these patients and decreased gene expression is associated with reduced symptoms. Confirmation of the achievement of this study requires further research. Key words: Schizophrenia, Thioredoxin, Risperidone

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Single-Nucleotide Variation Spectrum of the Factor XI Gene in Women with Heavy Menstrual Bleeding

Blood ◽

10.1182/blood.v124.21.4213.4213 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4213-4213

Author(s):

Sophie Wiewel-Verschueren ◽

André B Mulder ◽

Karina Meijer ◽

René Mulder

Keyword(s):

Heavy Chain ◽

Light Chain ◽

Heavy Menstrual Bleeding ◽

Menstrual Bleeding ◽

Control Group ◽

Single Nucleotide Variants ◽

Factor Xi ◽

Single Nucleotide ◽

Hardy Weinberg Equilibrium ◽

Bleeding Score

Abstract Introduction: In plasma, factor XI (FXI) circulates as a homodimeric precursor of a serine protease (FXIa), which plays an essential role in the contact activation of coagulation through the conversion of FIX to FIXa in a calcium-dependent manner. Each FXI monomer contains a heavy chain and a light chain that are joined together by disulphide bonds. The heavy chain contains all four apple domains and the light chain contains the serine protease domain. The fourth apple domain is necessary for dimerization. The gene for FXI is located on the long arm of chromosome 4 (4q35) and contains 15 exons and 14 introns. To date, more than 240 mutations have been reported (http://www.factorxi.org). The prevalence of FXI deficiency in Caucasians is reported as low, but might be underestimated. Women with low FXI levels (<70%) are prone to excessive bleeding during menstruation. However, bleeding manifestations are not well correlated with the plasma FXI levels and bleeding episodes can vary widely among patients with similar low FXI levels. In our previous study (Knol et al, AJOG, 2013), we found 4% FXI deficiency (< 70%) in unselected Dutch women with heavy menstrual bleeding (HMB). We also found that patients had significantly longer APTT compared to controls (26.5 vs 25.0 sec; p=0.001), despite higher levels of factor VIII. This turned out to be caused by lower median levels of FXI (100 vs 124 IU/dL; p<0.001). These lower levels of FXI and increased bleeding tendency could be caused by the presence of specific single-nucleotide variants in the FXI gene in women with HMB. To our knowledge, systemic analysis of FXI gene variants in women with HMB has not been reported. Aim: to determine the single-nucleotide variants of the FXI gene in women with heavy menstrual bleeding. Methods: the study was approved by the Institutional Review Board of the University Medical Center of Groningen. Informed consent was obtained from all patients. We included patients referred for heavy menstrual bleeding (PBAC-score >100). We measured the Tosetto bleeding score by questionnaire. FXI activity levels were determined by an one-stage clotting assay (Siemens, Marburg, Germany). Reference interval was 65-150%. Direct sequencing analysis of all 15 exons and flanking introns of the factor XI gene was performed to detect single-nucleotide variants. Results were compared with the HapMap and 1000 genome database using the Fisher exact probability test on a 2x3 contingency table. In addition, we tested each single-nucleotide variant for Hardy-Weinberg equilibrium. Results: We included 49 patients. Median FXI level was 96% (range 61%-155%). We found 31 different single-nucleotide variants in 49 patients with HMB. Seven out of 31 could not be compared to the control group due to small numbers, unknown frequency, or absence from the database. From the remaining 24 single-nucleotide variants (Figure 1), two (rs925451 and rs2241817) showed a significant difference compared to the control group (P<0.01). These two single-nucleotide variants showed also a significant departure from the Hardy-Weinberg equilibrium (P<0.01). There was no correlation between the number of single-nucleotide variants and the FXI level, PBAC-score or bleeding score. Conclusions: Our study provides a detailed analysis of single-nucleotide variants of the factor XI gene. Among these 31 variants, rs925451 and rs2241817 seem to be associated with heavy menstrual bleeding.Overall, these data may serve as reference group for future studies on the molecular background of factor XI deficiency and heavy menstrual bleeding. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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Immunomodulatory and Anti-Inflammatory Potentials of Trigona Honey in the Therapy and Prevention against Respiratory Infection in Wistar Rats

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11i3.2385 ◽

2020 ◽

Vol 11 (3) ◽

pp. 2955-2962

Author(s):

Ibrahim Khaled Al-kafaween ◽

Abu Bakar Mohd Hilmi ◽

Mohamed M. Soliman

Keyword(s):

Respiratory Infection ◽

Wistar Rats ◽

Serum Levels ◽

Negative Control Group ◽

Treated Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Gene Expressions ◽

Anti Inflammatory

Trigona honey (TH) is well known for its therapeutic characteristics. To date, the study of Trigona honey as a prophylactic or immune booster prior to the bacterial infection of the invivo model is not well covered. This study aims to investigate anti-inflammatory and immune activities in Wistar rats infected with respiratory infection following with Trigona honey. 25 Wistar rats were assigned to possitive groups, negative control group, positive control group was fed TH (5 g / kg body weight) orally, the untreated group was infected with Staphylococcus aureus to induce respiratory infection, the treated group has been infected with S. aureus followed by treatment with TH at a dose of 1.5 ×108 CFU / mL and the preventive group ingested TH one week before S. aureus infection. Blood was obtained for biochemical analysis. Lung tissues have been collected for molecular examination. The results showed a significant decrease in serum levels of ALT, AST, urea and creatinine in the preventive and treated groups, serum IgG increased significantly (P<0.05) in the preventive and treated groups, IFN-y increased in the preventive group while decreased in the treated group, and IL-8 increased in the treated group while decreased in the preventive group. The mRNA expression of AGP is up-regulated in the positive control, preventive and treated groups. The α2-MG, TNF-α , and mRNA expressions showed lower regulation after administration of TH in preventive and treated groups. The results show the ability of TH to counteract immune and inflammatory changes in serum levels and gene expressions.

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Evaluation of EOS Gene Expression and IL-6 Serum Levels in Iraqi Patients with Psoriasis

Al-Rafidain Journal of Medical Sciences ( ISSN: 2789-3219 ) ◽

10.54133/ajms.v1i.36 ◽

2021 ◽

Vol 1 ◽

pp. 89-93

Author(s):

Shawq Al-Naqqash ◽

Mohammed Mahdi Jawad ◽

Zainab Thamer Showait Al-Asady ◽

Sarmed Adnan Abdulrazaq

Keyword(s):

Gene Expression ◽

Topical Therapy ◽

Critical Role ◽

Serum Levels ◽

Pleiotropic Effect ◽

Control Group ◽

Dermatology Clinic ◽

Healthy Donors ◽

Significant Difference ◽

Soluble Mediator

Background: EOS (encoded by the IKZF4 gene) is a member of the zinc finger transcription factor IKaros family, and plays a critical role in Treg suppressor functions, and maintaining Treg stability. IL-6 is a soluble mediator with a pleiotropic effect on inflammation, immune response, and hematopoiesis. Aim: To estimate serum IL-6 level and EOS gene expression in Iraqi patients with psoriasis. Method: Twenty-two patients with psoriasis (8 females, 14 males) with age ranged 18-72 years, were recruited from Baghdad Teaching Hospital, Dermatology Clinic, Baghdad, and 24 healthy donors. The serum levels of IL-6 by ELISA and the gene expression of IKZF4 (EOS gene) by RT-qPCR technique. Results: The results showed a non-significant difference in the level of IL-6 in those treated with topical therapy and others treated with Etanercept compared to control. A non-significant increase in patients treated with topical therapy was reported compared to patients treated with Etanercept. There was a higher significant percentage of IKZF4 gene expression folding in psoriasis patients treated with Etanercept compared to control group, while no significant differences reported between patients treated with topical therapy, Etanercept, and the control group. Conclusion: Activation of Regulatory T cells (Tregs) with Etanercept enhances EOS expression and decreases IL-6 production more than topical treatment in patients with psoriasis.

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The effect of Lactobacillus johnsonii Ncc533 (La1) on the balance of Th1/Th2 cells in BALB/c mice

Clinical & Investigative Medicine ◽

10.25011/cim.v34i4.15369 ◽

2011 ◽

Vol 34 (4) ◽

pp. 254 ◽

Cited By ~ 4

Author(s):

Junli Yang ◽

Wen Li ◽

Ruopeng Sun ◽

Baomin Li

Keyword(s):

Flow Cytometry ◽

Immune Response ◽

Serum Levels ◽

Treated Group ◽

Specific Ige ◽

Th2 Cells ◽

Control Group ◽

Lactobacillus Johnsonii ◽

Asthma Model ◽

Ifn Γ

Purpose: To determine the effect of Lactobacillus johnsonii Ncc533 (La1) on Th1/Th2 balance, the production of IL-4 and IFN-γ by splenocytes was evaluated following its administration to mice from newborn to adult. Changes in IL-4 and IFN-γ expression and serum levels of OVA-specific-IgE were then investigated in an asthma model. Methods: Using flow cytometry (FCM) and ELISA, the percentage of IL-4 and IFN-γ expressing splenocytes and serum levels of OVA-specific-IgE were measured in different groups of mice. Results: The percentages of IL-4 and IFN-γ expressing splenocytes in the offspring and in the adults of the La1-treated group were not significantly different when compared with the water-treated group. In the asthma model, the percentages of IL-4 expressing cells and the serum levels of OVA-specific-IgE in the La1-treated and water-treated group were significantly increased compared with those in the control group. The percentage of IFN-γ expressing cells was significantly lower in the La1-treated and water-treated groups. The percentage of IL-4 expressing cells and the serum levels of OVA-specific-IgE in the La1-treated group were significantly lower compared with those in the water-treated group, whereas the percentage of IFN-γ expressing cells was significantly higher. Conclusion: Administration of La1 had no effect on the immune system from the neonate to the adult in the normal mice. It did, however, significantly alter the percentages of IL-4 or IFN-γ expressing CD4+ T lymphocytes in the asthma model, suggesting that administration of La1 might regulate the immune response.

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Local and Systemic Up-regulation of TNFα, IL-1β and IL-6 in Mice Intratracheally Inoculated with Porphyromonas gingivalis

Acta Veterinaria Brno ◽

10.2754/avb200877030377 ◽

2008 ◽

Vol 77 (3) ◽

pp. 377-385 ◽

Cited By ~ 2

Author(s):

Z. Pavlica ◽

A. Nemec ◽

A. Nemec-Svete ◽

D. Eržen ◽

D. A. Crossley ◽

...

Keyword(s):

Immune Response ◽

Immune Responses ◽

Porphyromonas Gingivalis ◽

Total Protein ◽

Protein Concentration ◽

Serum Levels ◽

Treated Group ◽

Control Group ◽

Total Protein Concentration ◽

Sterile Phosphate Buffer

The objective of the study was to find whether a single intratracheal inoculation with live Porphyromonas gingivalis ATCC 33277 influences local and systemic inflammatory and immune responses in mice.Twelve-week-old BALB/c mice were intratracheally inoculated with 2.9 × 109 CFU P. gingivalis ATCC 33277 diluted in 40 μl sterile phosphate buffer (treated group) or with sterile PBS (control group). The animals were sacrificed 2, 6, 24, 72 and 168 h after inoculation. TNFα, IL-1β, IL-6 and total protein concentrations were measured in the serum, lungs and kidneys. Six hours after P. gingivalis inoculation, TNFα concentration was significantly increased in serum (p = 0.02) and kidneys (p = 0.04), but in the lungs TNFα production was enhanced already 2 h (p < 0.0001) after inoculation, reaching the peak after 6 h (p < 0.0001). The IL-1β concentration was also significantly increased in serum after 2 h (p = 0.006), remaining significantly elevated up to 3 days (p ≤ 0.0001) after inoculation. In lungs IL-1β levels were significantly increased 6 and 24 h (p < 0.0001) and in kidneys 24 h (p < 0.0001) and 168 h (p = 0.01) after inoculation. The IL-6 concentration was significantly increased in serum after 72 and 168 h (p < 0.0001). However, IL-6 was significantly increased in lungs after 6 h (p < 0.0001), remaining elevated until 72 h and in kidneys 2 and 6 h (p < 0.0001) after inoculation. Significantly increased total protein concentration was detected in kidneys 6 and 24 h (p < 0.0001) after inoculation. These results suggest that a single intratracheal inoculation with P. gingivalis stimulates the local and systemic inflammatory and immune response, as shown by increased tissue and serum levels of proinflammatory cytokines.

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In-depth comparative toxicogenomics of glyphosate and Roundup herbicides: histopathology, transcriptome and epigenome signatures, and DNA damage

10.1101/2021.04.12.439463 ◽

2021 ◽

Author(s):

Robin Mesnage ◽

Mariam Ibragim ◽

Daniele Mandrioli ◽

Laura Falcioni ◽

Fiorella Belpoggi ◽

...

Keyword(s):

Oxidative Stress ◽

Gene Expression ◽

Dna Methylation ◽

European Union ◽

Dna Damage ◽

Protein Misfolding ◽

Control Group ◽

The European Union ◽

Dna Methylation Profiling ◽

Methylation Profiling

Background. Health effects from exposure to glyphosate-based herbicides is an intense matter of debate. Toxicity including genotoxicity of glyphosate alone has been repeatedly tested over the last 40 years. Contrastingly, few studies have conducted comparative investigations between glyphosate and its commercial herbicide formulations, such as Roundup. We thus performed the first in-depth comparative toxicogenomic evaluation of glyphosate and a typical European Union Roundup formulation by determining alterations in transcriptome and epigenome profiles. Methods. Glyphosate and the European Union reference commercial formulation Roundup MON 52276 (both at 0.5, 50, 175 mg/kg bw/day glyphosate equivalent concentration) were administered to rats in a subchronic 90-day toxicity study. Standard clinical biochemistry and kidney and liver histopathology was performed. In addition, transcriptomics and DNA methylation profiling of liver and selective gene expression analysis of kidneys was conducted. Furthermore, a panel of six mouse embryonic reporter stem cell lines validated to identify carcinogenic outcomes (DNA damage, oxidative stress, and protein misfolding) were used to provide insight into the mechanisms underlying the toxicity of glyphosate and 3 Roundup formulations. Results. Histopathology and serum biochemistry analysis showed that MON 52276 but not glyphosate treatment was associated with a statistically significant increase in hepatic steatosis and necrosis. Similar lesions were also present in the liver of glyphosate-treated groups but not in the control group. MON 52276 altered the expression of 96 genes in liver, with the most affected biological functions being TP53 activation by DNA damage and oxidative stress as well as the regulation of circadian rhythms. The most affected genes in liver also had their expression similarly altered in kidneys. DNA methylation profiling of liver revealed 5,727 and 4,496 differentially methylated CpG sites between the control group and the group of rats exposed to glyphosate and MON 52276, respectively. Direct DNA damage measurement by apurinic/apyrimidinic lesion formation in liver was increased with glyphosate exposure. Mechanistic evaluations showed that two Roundup herbicides but not glyphosate activated oxidative stress and misfolded protein responses. Conclusions. Taken together, the results of our study show that Roundup herbicides are more toxic than glyphosate, activating mechanisms involved in cellular carcinogenesis and causing gene expression changes reflecting DNA damage. This further highlights the power of high-throughput omics methods to detect metabolic changes, which would be missed by relying solely on conventional biochemical and histopathological measurements. Our study paves the way for future investigations by reporting a panel of gene expression changes and DNA methylation sites, which can serve as biomarkers and potential predictors of negative health outcomes resulting from exposure to glyphosate-based herbicides.

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