Folic acid reduces methomyl insecticide damage to testicular cells by altering the DNA methylation environment

doi:10.21203/rs.3.rs-1332534/v1

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Folic acid reduces methomyl insecticide damage to testicular cells by altering the DNA methylation environment

https://doi.org/10.21203/rs.3.rs-1332534/v1

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Methomyl is a carbamate insecticide with confirmed testicular toxicity. This study intends to observe the damage of methomyl on testicular cells and the protective effect of folic acid through in vitro experiments. We treated the GC-1 spermatogonia, TM4 Sertoli cells, and TM3 Leydig cells with Methomyl (0, 250, 500, and 1000 μM) with or not folic acid (0, 10, 100, and 1000 nM) for 24h. It was found that folic acid attenuated the testicular cytotoxicity of methomyl in a dose-dependent manner. In spermatogonia, methomyl significantly inhibited the expression of proliferation genes Ki67 and PCNA at 1000 μM, and increased the expression of apoptosis genes Caspase3 and Bax at each dose, while folic acid could reverse the change caused by methomyl. In Sertoli cells, methomyl dose-dependently inhibited the expression of blood-testis barrier function genes TJP1, Cx43, and N-cadherin, but did not affect Occludin and E-cadherin, while folic acid could significantly up-regulate the expression of TJP1, Cx43, and Occludin. In Leydig cells, methomyl dose-dependently inhibited the expression of steroid synthase P450scc, StAR, Hsd3b1 and down-regulated the level of testosterone, but did not affect Cyp17a1 and Hsd17b1, while folic acid could significantly up-regulate the expression of P450scc, StAR, Hsd3b, Cyp17a1, and Hsd17b1, and reverse testosterone level downtrend. Simultaneously, methomyl did not affect the expression of Dnmt1, Dnmt3A, Dnmt3B genes in testicular cells, while folic acid dose-dependently up-regulated the expression of Dnmt1, Dnmt3A, and Dnmt3B in spermatogonia, Dnmt1 and Dnmt3B in Sertoli cells, and Dnmt1 and Dnmt3A in Leydig cells. In conclusion, folic acid attenuated the damage of methomyl on testicular spermatogonia, Sertoli cells, and Leydig cells by altering the DNA methylation environment.

Methomyl

folic acid

Leydig cell

Sertoli cell

spermatogonia

Methomyl is a carbamate insecticide widely used in the foliar treatment of vegetables, fruits, crops, cotton, and commercial ornamentals (Jiang, Zhang et al. 2020). Human exposure to methomyl in the environment through occupational handling, fly spraying, and consumption of related food (Van Scoy, Yue et al. 2013). Also, due to its high water solubility (57.9 g/L at 25°C) and low adsorption to the soil, methomyl can seep into groundwater and pollute drinking water resources (Meng, Qiu et al. 2016). Methomyl is classified as a restricted-use pesticide due to its toxicity to non-target species (Van Scoy, Yue et al. 2013). The WHO (World Health Organization), EPA (Environmental Protection Agency), and EC (European Commission) has also declared methomyl to be a toxic and hazardous pesticide (Jiang, Zhang et al. 2020). However, methomyl is still widely used as a pesticide in some areas (Liu, Gao et al. 2017). Therefore, Methomyl still has potential harm to human health.

Methomyl's effect on target organisms is due to the inhibition of acetylcholinesterase. Still, it is also an inhibitor of enzymes such as superoxide dismutase and glutathione S-transferase, which catalyze the elimination of reactive oxygen species (ROS) (Mansour, Mossa et al. 2009). Methomyl has been identified to induce cellular DNA damage and apoptosis in vitro (Guanggang, Diqiu et al. 2013). In recent years, studies have found that methomyl at 20 and 200 μg/L treated tilapia for 30 days significantly enhanced Caspase-8 activity, induced apoptosis (Meng, Liu et al. 2019), and led to changes in testicular sex hormones and vitellogenin levels (Meng, Qiu et al. 2016). It is suggested that methomyl has severe testicular toxicity.

Non-enzymatic antioxidants, usually derived from the diet, help maintain cellular redox homeostasis. Folic acid, a water-soluble vitamin, has antioxidant activity comparable to vitamins C and E (Henderson, Tai et al. 2018). In vivo, folate acid in conjunction with vitamins B12 and B6 is involved in nucleotide synthesis, the regeneration of methionine from homocysteine, and oxidation and reduction of one-carbon units required for normal cellular function (Bailey and Gregory 1999). The study found that folic acid could reduce methomyl testicular toxicity and prevent tissue damage by directly scavenging reactive oxygen species and indirectly enhancing cellular redox homeostasis (Shalaby, El Zorba et al. 2010, Sakr, Hassanien et al. 2018).

Due to the lack of in vitro experiments on the damage of methomyl to the testis. In this study, we evaluated the damage of methomyl on spermatogonia, Sertoli cells, and Leydig cells in vitro and further observed the protective effect of folic acid on them.

2.1 Chemicals and reagents

Methomyl and folic acid were obtained from Sigma (St. Louis, MO). The CellTiter96® AQueous cell proliferation assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; MTS) was from Promega (Madison, WI, USA). Mouse iodine (125I) testosterone radioimmunoassay kits (S1094093) were purchased from the North Institute of Biological Technology (Beijing, China). The total RNA TRIzol kit was purchased from Omega Bio-Tek (Doraville, Georgia, USA). Reverse transcription and quantitative real-time PCR (RT-qPCR) kits (Q223) were purchased from Takara Biotechnology Co., Ltd. (Dalian, China). The primers were synthesized by TIANYIHUIYUAN Biotechnology Co., Ltd. (Wuhan, China). The other reagents for experiments were of analytical grade.

2.2 Cell culture and treatment

GC-1 cells (mouse testis spermatogonia), TM4 cells (mouse testis Sertoli cells), and TM3 cells (mouse testis Leydig cells) were acquired from Procell Life Science & Technology Co., Ltd. (Wuhan, China). Cells were cultured in DMEM/F12 medium, supplemented with 10% HS, and 1% penicillin-streptomycin at 37 °C in 5% CO2 in a humidified incubator. The medium was changed every 2–3 days. This study treated cells with 0, 250, 500, and 1000 μM Methomyl for 24 h to explore its damaging effects on testicular cells (Guanggang, Diqiu et al. 2013, Yun, Huang et al. 2017, Saquib, Siddiqui et al. 2021). Subsequently, we treated the cells with 1000 μM Methomyl and 0, 10, 100, and 1000 nM folic acid for 24 h to observe the protective effect of folic acid on testicular cells (Carletti, Correia-Branco et al. 2018).

2.3 Cytotoxicity testing

Cytotoxicity was assessed using the MTS assay. The GC-1 cells, TM3 cells, and TM4 cells were seeded at a concentration of 2 × 10⁵ cells/mL, 150 μL/well into a 96-well tissue culture plate. After the cells were cultured with Methomyl (0, 250, 500, and 1000 μM) with or not folic acid (0, 10, 100, and 1000 nM) for 24h, 20 μL of MTS reagent was added into each well, and the absorbance was assessed at 490 nm with a microplate reader after 1h incubation with MTS.

2.4 Measurement of testosterone level

The testosterone concentrations were measured by enzyme-linked immunosorbent assay (ELISA) kits. All assay procedures followed the manufacturer protocol as our descriptions.

2.5 Total RNA extraction, reverse transcription and RT-qPCR

Total RNA was isolated from cells using TRIzol Reagent following the manufacturer's protocol. The total RNA was reverse transcribed using a first-strand cDNA synthesis kit. Then, RT-qPCR was performed using an SYBR Green qPCR Master Mix Kit and ABI StepOnePlus cycler (Applied Biosystems, Foster City, CA, USA). The mouse primer sequences for the genes used in this study are shown in Table 1. The cycle threshold (Ct) was detected. The relative expression of genes was determined using the 2^△△Ct method with normalization to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression and used as a quantitative control. Primers of target genes are presented in Table 1.

2.6 Statistical analysis

SPSS 19 (SPSS Science Inc., Chicago, Illinois) and Prism 8.0 (Graph Pad Software, La Jolla, CA, USA) was used to analyze data. Quantitative data are expressed as the mean ± S.E.M., a two-tailed Student's t-test was used for comparisons between control and treatment groups. And for studies involving more than two groups, data were evaluated with one-way analysis of variance (ANOVA) followed by Tukey's post-hoc-test. Statistical significance was defined as P<0.05.

3.1 Effects of methomyl and folic acid on testicular cytotoxicity

First, we observed the effects of methomyl (0, 250, 500, and 1000 μM) on the cytotoxicity of GC-1 spermatogonia, TM4 Sertoli cells, and TM3 Leydig cells after 24 h treatment. The results of MTS showed that the absorbance value of methomyl-treated cells decreased, and the cytotoxicity increased significantly (P<0.01, Fig 1A-C). Further, we co-treated cells with methomyl (1000 μM) and folic acid (0, 10, 100, and 1000 nM) and found that folic acid attenuated the cytotoxicity of methomyl to testicular cells in a dose-dependent manner (P<0.05, P<0.01, Fig 1D-F).

3.2 Effects of methomyl and folic acid on proliferation and apoptosis in GC-1 spermatogonia

The results showed that methomyl at 1000 μM significantly inhibited the expressions of proliferation-related genes Ki67 and PCNA, and increased the expressions of apoptosis-related genes Caspase3 and Bax at each dose (P<0.05, P<0.01, Fig 2A-D). The combination of folic acid and methomyl can significantly up-regulate Ki67, PCNA, and down-regulate the expressions of Caspase3 and Bax (P<0.05, P<0.01, Fig 2E-H). It was suggested that methomyl resulted in the inhibition of spermatogonia proliferation and enhanced apoptosis, while folic acid attenuated the damage of methomyl to it.

3.3 Effects of methomyl and folic acid on blood-testis barrier function in TM4 Sertoli cells

We observed the effect of methomyl and folic acid on the blood-testis barrier function of TM4 Sertoli cells. The results showed that methomyl dose-dependently inhibited the expression of blood-testis barrier function genes TJP1, Cx43, and N-cadherin, but did not affect the expression of Occludin and E-cadherin (P<0.05, P<0.01, Fig 3A-E). It was further found that folic acid can significantly up-regulate the expression of TJP1, Cx43, and Occludin (P<0.05, P<0.01, Fig 3F-J). It is suggested that methomyl inhibits Sertoli cell blood-testis barrier function, and folic acid can partially improve the damage caused by methomyl.

3.4 Effects of methomyl and folic acid on testosterone synthesis in TM3 Leydig cells

We observed the effects of methomyl and folic acid on testosterone synthesis in TM3 Leydig cells. The results showed that methomyl dose-dependently inhibited the expression of steroid synthase P450scc, StAR, and Hsd3b1, but did not affect the expression of Cyp17a1 and Hsd17b1 (P<0.01, Fig 4A-E), and down-regulated the level of testosterone in Leydig cells (P<0.01, Fig 4F). It was further found that folic acid could significantly up-regulate the expressions of P450scc, StAR, Hsd3b, Cyp17a1, and Hsd17b1 (P<0.05, P<0.01, Fig 4D-K), and reverse the downward trend of testosterone levels (P<0.05, P<0.01, Fig 4L). It is suggested that methomyl inhibits the synthesis of testosterone in Leydig cells, while folic acid can attenuate the damage of methomyl to it.

3.5 Effects of methomyl and folic acid on DNA methylation environment in testicular cells

Finally, we explored the effects of methomyl and folic acid on DNA methylase in testicular cells. The results showed that 1000 μM methomyl did not affect the expression of Dnmt1, Dnmt3A and Dnmt3B genes in testicular cells, while folic acid dose-dependently up-regulated the expression of Dnmt1, Dnmt3A and Dnmt3B in spermatogonia, Dnmt1 and Dnmt3B in Sertoli cells, and Dnmt1 and Dnmt3A in Leydig cells (P<0.05, P<0.01, Fig 5A-I). It is suggested that folic acid attenuates the damage of methomyl on testicular cells by changing the DNA methylation environment.

Pesticides have adverse effects on cells. A study systematically tested the cytotoxic potential of 13 commonly used agricultural pesticides on three human HepG2, Hek293, HeLa cells, and three insect Tn5B1-4, Sf-21, and Drosophila S2 cells and found that some pesticides promoted cell proliferation, while some pesticides have a significant inhibitory effect on cell viability (Yun, Huang et al. 2017). They showed that even 24h exposure to methomyl at 20 μg/ml produced little cytotoxicity to the tested human and insect cells (Yun, Huang et al. 2017). However, human umbilical vein endothelial cells exposed to methomyl for 24 h exhibited significant proliferation inhibition at higher concentrations (500 and 1000 μM) (Saquib, Siddiqui et al. 2021). It is suggested that different cells have different susceptibility to the same insecticide. Folic acid, a member of the vitamin B9 family, is essential for the biosynthesis of nucleotides, amino acids, and S-adenosyl-l-methionine (Li, Ma et al. 2019, Koohpeyma, Goudarzi et al. 2020, Wusigale, Hu et al. 2020). Animal studies have demonstrated that methomyl reduces testosterone levels and impairs sperm quality in male rats, while folic acid improves methomyl's impairment of reproductive performance (Shalaby, El Zorba et al. 2010, Sakr, Hassanien et al. 2018). In this study, we found that 500 and 1000 μM methomyl significantly increased cytotoxicity after 24h in vitro treatment of GC-1 spermatogonia, TM4 Sertoli cells, and TM3 Leydig cells, while folic acid attenuated the testis cytotoxicity of methomyl in a dose-dependent manner.

The testis is composed of seminiferous epithelium and mesenchyme (Hai, Hou et al. 2014). The former is a unique site of spermatogenesis, mainly composed of spermatogonia and Sertoli cells (Wen, Tang et al. 2016), while the latter is the place where mesenchymal cells synthesize and secrete androgens (Zhou, Wu et al. 2019). It is known that spermatogenesis begins with the proliferation of spermatogonia. Spermatogonia continue to self-renew and proliferate throughout adult life to maintain stem cell reserves while providing cells for the spermatogenesis cycle (Manku and Culty 2015). Therefore, the proliferation and apoptosis of spermatogonia are crucial for spermatogenesis. In the testis, the blood-testis barrier composed of a variety of structural proteins (such as TJP1, Cx43, N-cadherin, Occludin, E-cadherin, etc.) between adjacent Sertoli cells, can maintain normal spermatogenesis by participating in the construction of the spermatogenic microenvironment, providing an immune barrier, and conferring cell polarity in the seminiferous epithelium (Cheng and Mruk 2012). Meanwhile, steroidogenesis in Leydig cells involves the conversion of cholesterol to testosterone through several intermediate steroids, a process regulated and catalyzed by P450scc, StAR, Hsd3b1, CYP17a1, and HSD17b1 (Hong, Chen et al. 2016, Zhao, Xiao et al. 2021). We found that in spermatogonia, methomyl at 1000 μM significantly inhibited the expression of proliferation genes Ki67 and PCNA, and increased the expression of apoptosis genes Caspase3 and Bax at various doses, while folic acid could reverse the changes in related gene expression caused by methomyl; In Sertoli cells, methomyl inhibited the expression of blood-testis barrier function genes TJP1, Cx43, N-cadherin in a dose-dependent manner, but did not affect the expression of Occludin and E-cadherin, while folic acid significantly up-regulated the expression of TJP1, Cx43, and Occludin; in Leydig cells, methomyl dose-dependently inhibited the expression of steroid synthase P450scc, StAR, Hsd3b1 and down-regulated the level of testosterone, but did not affect Cyp17a1 and Hsd17b1, while folic acid could significantly up-regulate the expression of P450scc, StAR, Hsd3b, Cyp17a1, and Hsd17b1, and reverse testosterone level downtrend. All in all, Methomyl can damage the cell function of testicular spermatogonia, Sertoli cells, and Leydig cells, while folic acid can reduce this damage.

Epigenetic information encoded by 5-methylcytosine (5mC) plays a key role in mammalian development and human disease. De novo production and maintenance of 5mC is regulated by DNA methyltransferases, namely DNMT1, DNMT3A, and DNMT3B. DNA methylation in mammals is thought to be critical for essential functions, including gene silencing leading to genomic imprinting, X chromosome inactivation, and repression of transposable elements (Liu, Liu et al. 2020). Folic acid is known to be critical in the process of supplying methyl groups for methylation in one-carbon metabolism, and low folate status is associated with an increased risk of cardiovascular disease, various cancers, and neural tube defects (Crider, Yang et al. 2012). Studies have found that folic acid can restore hepatic triglyceride accumulation induced by a high-fat sucrose diet by altering the methylation pattern of fatty acid synthase promoter (Cordero, Gomez-Uriz et al. 2013), inhibiting amyloid β-peptide production by modulating the activity of DNMTs in N2a-APP cells (Li, Jiang et al. 2015), and also alleviate oxidative stress-induced apoptosis in vivo and in vitro by increasing DNMTs activity and VPO1 promoter methylation level (Luo, Zhang et al. 2013). Thus, by altering DNA methylation, the use of folic acid can be used as a treatment modality for many diseases. In this paper, we found that methomyl did not affect the expression of Dnmt1, Dnmt3A, Dnmt3B genes in testicular cells, while folic acid dose-dependently up-regulated the expression of Dnmt1, Dnmt3A and Dnmt3B in spermatogonia, Dnmt1, and Dnmt3B in Sertoli cells, and Dnmt1 and Dnmt3A in Leydig cell. It is suggested that folic acid attenuates the damage of methomyl to testicular cells by changing the DNA methylation environment.

We confirmed the damage of methomyl to testicular spermatogonia, Sertoli cells, and Leydig cells, and proposed the protective mechanism of folic acid attenuating methomyl on testicular cell damage by changing the DNA methylation environment. Considering the serious harm of methomyl to human society, further mechanistic studies using animal models are urgently needed. This study provides new insights into the toxic effects and protective mechanisms of methomyl.

Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

Competing Interests

The authors have no relevant financial or non-financial interests to disclose

Author Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by [Mengxi Lu] and [Yi Liu]. The first draft of the manuscript was written by [Yi Liu] and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Disclosure of potential conflicts of interest

There are no potential conflicts of interest in this research.

Research involving Human Participants and/or Animals

This study does not involve studies of human participants and animals.

Informed consent

This study does not involve studies of human participants.

Availability of data and materials

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Bailey, L. B. and J. F. Gregory, 3rd (1999). "Folate metabolism and requirements." J Nutr129(4): 779-782.
Carletti, J. V., A. Correia-Branco, C. R. Silva, N. Andrade, L. O. P. Silva and F. Martel (2018). "The effect of oxidative stress induced by tert-butylhydroperoxide under distinct folic acid conditions: An in vitro study using cultured human trophoblast-derived cells." Reprod Toxicol77: 33-42.
Cheng, C. Y. and D. D. Mruk (2012). "The blood-testis barrier and its implications for male contraception." Pharmacol Rev64(1): 16-64.
Cordero, P., A. M. Gomez-Uriz, J. Campion, F. I. Milagro and J. A. Martinez (2013). "Dietary supplementation with methyl donors reduces fatty liver and modifies the fatty acid synthase DNA methylation profile in rats fed an obesogenic diet." Genes Nutr8(1): 105-113.
Crider, K. S., T. P. Yang, R. J. Berry and L. B. Bailey (2012). "Folate and DNA methylation: a review of molecular mechanisms and the evidence for folate's role." Adv Nutr3(1): 21-38.
Guanggang, X., L. Diqiu, Y. Jianzhong, G. Jingmin, Z. Huifeng, S. Mingan and T. Liming (2013). "Carbamate insecticide methomyl confers cytotoxicity through DNA damage induction." Food Chem Toxicol53: 352-358.
Hai, Y., J. Hou, Y. Liu, Y. Liu, H. Yang, Z. Li and Z. He (2014). "The roles and regulation of Sertoli cells in fate determinations of spermatogonial stem cells and spermatogenesis." Semin Cell Dev Biol29: 66-75.
Henderson, A. M., D. C. Tai, R. E. Aleliunas, A. M. Aljaadi, M. B. Glier, E. E. Xu, J. W. Miller, C. B. Verchere, T. J. Green and A. M. Devlin (2018). "Maternal folic acid supplementation with vitamin B12 deficiency during pregnancy and lactation affects the metabolic health of adult female offspring but is dependent on offspring diet." FASEB J32(9): 5039-5050.
Hong, J., F. Chen, X. Wang, Y. Bai, R. Zhou, Y. Li and L. Chen (2016). "Exposure of preimplantation embryos to low-dose bisphenol A impairs testes development and suppresses histone acetylation of StAR promoter to reduce production of testosterone in mice." Mol Cell Endocrinol427: 101-111.
Jiang, W., C. Zhang, Q. Gao, M. Zhang, J. Qiu, X. Yan and Q. Hong (2020). "Carbamate C-N Hydrolase Gene ameH Responsible for the Detoxification Step of Methomyl Degradation in Aminobacter aminovorans Strain MDW-2." Appl Environ Microbiol87(1).
Koohpeyma, H., I. Goudarzi, M. Elahdadi Salmani, T. Lashkarbolouki and M. Shabani (2020). "Folic Acid Protects Rat Cerebellum Against Oxidative Damage Caused by Homocysteine: the Expression of Bcl-2, Bax, and Caspase-3 Apoptotic Genes." Neurotox Res37(3): 564-577.
Li, W., M. Jiang, S. Zhao, H. Liu, X. Zhang, J. X. Wilson and G. Huang (2015). "Folic Acid Inhibits Amyloid beta-Peptide Production through Modulating DNA Methyltransferase Activity in N2a-APP Cells." Int J Mol Sci16(10): 25002-25013.
Li, W., Y. Ma, Z. Li, X. Lv, X. Wang, D. Zhou, S. Luo, J. X. Wilson and G. Huang (2019). "Folic Acid Decreases Astrocyte Apoptosis by Preventing Oxidative Stress-Induced Telomere Attrition." Int J Mol Sci21(1).
Liu, H. Y., S. M. Liu and Y. Z. Zhang (2020). "Maternal Folic Acid Supplementation Mediates Offspring Health via DNA Methylation." Reprod Sci27(4): 963-976.
Liu, Y., Y. Gao, G. Liang and Y. Lu (2017). "Chlorantraniliprole as a candidate pesticide used in combination with the attracticides for lepidopteran moths." PLoS One12(6): e0180255.
Luo, S., X. Zhang, M. Yu, H. Yan, H. Liu, J. X. Wilson and G. Huang (2013). "Folic acid acts through DNA methyltransferases to induce the differentiation of neural stem cells into neurons." Cell Biochem Biophys66(3): 559-566.
Manku, G. and M. Culty (2015). "Mammalian gonocyte and spermatogonia differentiation: recent advances and remaining challenges." Reproduction149(3): R139-157.
Mansour, S. A., A. T. Mossa and T. M. Heikal (2009). "Effects of methomyl on lipid peroxidation and antioxidant enzymes in rat erythrocytes: in vitro studies." Toxicol Ind Health25(8): 557-563.
Meng, S., L. Qiu, G. Hu, L. Fan, C. Song, Y. Zheng, W. Wu, J. Qu, D. Li, J. Chen and P. Xu (2016). "Effects of methomyl on steroidogenic gene transcription of the hypothalamic-pituitary-gonad-liver axis in male tilapia." Chemosphere165: 152-162.
Meng, S. L., T. Liu, X. Chen, L. P. Qiu, G. D. Hu, C. Song, L. Fan, Y. Zheng, J. Z. Chen and P. Xu (2019). "Effect of Chronic Exposure to Methomyl on Tissue Damage and Apoptosis in Testis of Tilapia (Oreochromis niloticus) and Recovery Pattern." Bull Environ Contam Toxicol102(3): 371-376.
Sakr, S., H. Hassanien, M. J. Bester, S. Arbi, A. Sobhy, H. El Negris and V. Steenkamp (2018). "Beneficial effects of folic acid on the kidneys and testes of adult albino rats after exposure to methomyl." Toxicol Res (Camb)7(3): 480-491.
Saquib, Q., M. A. Siddiqui, S. M. Ansari, H. A. Alwathnani, J. Musarrat and A. A. Al-Khedhairy (2021). "Cytotoxicity and genotoxicity of methomyl, carbaryl, metalaxyl, and pendimethalin in human umbilical vein endothelial cells." J Appl Toxicol41(5): 832-846.
Shalaby, M. A., H. Y. El Zorba and R. M. Ziada (2010). "Reproductive toxicity of methomyl insecticide in male rats and protective effect of folic acid." Food Chem Toxicol48(11): 3221-3226.
Van Scoy, A. R., M. Yue, X. Deng and R. S. Tjeerdema (2013). "Environmental fate and toxicology of methomyl." Rev Environ Contam Toxicol222: 93-109.
Wen, Q., E. I. Tang, X. Xiao, Y. Gao, D. S. Chu, D. D. Mruk, B. Silvestrini and C. Y. Cheng (2016). "Transport of germ cells across the seminiferous epithelium during spermatogenesis-the involvement of both actin- and microtubule-based cytoskeletons." Tissue Barriers4(4): e1265042.
Wusigale, L. Hu, H. Cheng, Y. Gao and L. Liang (2020). "Mechanism for Inhibition of Folic Acid Photodecomposition by Various Antioxidants." J Agric Food Chem68(1): 340-350.
Yun, X., Q. Huang, W. Rao, C. Xiao, T. Zhang, Z. Mao and Z. Wan (2017). "A comparative assessment of cytotoxicity of commonly used agricultural insecticides to human and insect cells." Ecotoxicol Environ Saf137: 179-185.
Zhao, L., Y. Xiao, C. Li, J. Zhang, Y. Zhang, M. Wu, T. Ma, L. Yang, X. Wang, H. Jiang, Q. Li, H. Zhao, Y. Wang, A. Wang, Y. Jin and H. Chen (2021). "Zearalenone perturbs the circadian clock and inhibits testosterone synthesis in mouse Leydig cells." J Toxicol Environ Health A84(3): 112-124.
Zhou, R., J. Wu, B. Liu, Y. Jiang, W. Chen, J. Li, Q. He and Z. He (2019). "The roles and mechanisms of Leydig cells and myoid cells in regulating spermatogenesis." Cell Mol Life Sci76(14): 2681-2695.

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Folic acid reduces methomyl insecticide damage to testicular cells by altering the DNA methylation environment

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