ABSTRACT
A widely used type II pyrethroid pesticide cypermethrin (CYP) is one of endocrine disrupting chemicals (EDCs) with anti-androgenic activity to induce male reproductive toxicology. However, the mechanisms have not been fully elucidated. This study was to explore the effects of CYP on apoptosis of mouse Sertoli cells (TM4) and the roles of endoplasmic reticulum (ER)-mitochondria coupling involving 1,4,5-trisphosphate receptor type1-glucose-regulated protein 75-voltage-dependent anion channel 1 (IP3R1-GRP75-VDAC1). TM4 were cultured with different concentrations of CYP. Flow cytometry, calcium (Ca2+) fluorescent probe, transmission electron microscopy and confocal microscopy, and western blot were to examine apoptosis of TM4, mitochondrial Ca2+, ER-mitochondria coupling, and expressions of related proteins. CYP was found to increase apoptotic rates of TM4 significantly. CYP was shown to significantly increase expressions of cleaved caspase-3, cleaved poly ADP-ribose polymerase (PARP). Concentration of mitochondrial Ca2+ was increased by CYP treatment significantly. CYP significantly enhanced ER-mitochondria coupling. CYP was shown to increase expressions of IP3R, Grp75 and VDAC1 significantly. We suggest that CYP induces apoptosis in TM4 cells by facilitating mitochondrial Ca2+ overload regulated by ER-mitochondria coupling involving IP3R1-GRP75-VDAC1. This study identifies a novel mechanism of CYP-induced apoptosis in Sertoli cells.
Subject(s)
HSP70 Heat-Shock Proteins , Membrane Proteins , Pyrethrins , Sertoli Cells , Mice , Animals , Male , Sertoli Cells/metabolism , Mitochondria , Endoplasmic Reticulum/metabolism , Pyrethrins/toxicity , Apoptosis , Calcium/metabolismABSTRACT
Cypermethrin (CYP) has been identified as one kind of endocrine-disrupting chemicals (EDCs) to induce male reproduction damage. This study aimed to investigate the effects and mechanisms of miR-30a-5p on CYP induced apoptosis of TM4 mouse Sertoli cells in vitro. In the present study, 0 µM, 10 µM, 20 µM, 40 µM and 80 µM CYP were used to treat TM4 cells for 24 h. The apoptosis of TM4 cells, the expression level of miR-30a-5p, the protein expressions and the interaction between miR-30a-5p and KLF9 were detected by flow cytometry, quantitative Real-Time PCR, Western blot and luciferase reporter assays. CYP induced apoptosis of TM4 cells, inhibited expression of miR-30a-5p in TM4 cells, and overexpression of miR-30a-5p partially recovered CYP induced cells apoptosis. Furthermore, KLF9 was a potential downstream target of miR-30a-5p predicted by publicly available databases. KLF9 expression level in TM4 cells was significantly elevated after treatment with CYP, and the induction was inhibited by miR-30a-5p mimics transfection. Meanwhile, dual-luciferase reporter assay demonstrated that miR-30a-5p directly targeted KLF9-3'UTR. Moreover, in the presence of CYP, the apoptosis regulator p53 expression was also increased in TM4 cells. Overexpression miR-30a-5p or down-regulation of KLF9 both attenuated the induction of CYP on p53 expression. Overall, the present study demonstrated that miR-30a-5p regulated CYP induced TM4 cells apoptosis by targeting KLF9/p53 axis.
Subject(s)
MicroRNAs , Animals , Mice , Male , MicroRNAs/genetics , Sertoli Cells/metabolism , Tumor Suppressor Protein p53/genetics , Cell Line, Tumor , Cell Proliferation , ApoptosisABSTRACT
Microplastics (MPs) have become an important global issue in recent years. However, MPs in the soil have received far less attention than water. Effective and nondestructive extraction of MPs is important for studying MPs in agricultural soils. This study uses different floatation solutions as experiments and uses MgCl2 as the floatation solution of the density extraction method. Five types of standard MPs (PE, PP, PS, PVC, and PET) are used as the objects of this experiment. The recovery of the two particle sizes was between 90.82% and 109.69%. The extracted standard MPs were then subjected to IR and Raman spectroscopic analysis, and the results showed that Raman spectroscopy was more suitable for the identification of the extracted MPs. Finally, this method collected and verified a vast number of soil samples and further analyzed the abundance and characteristics of the collected MPs.
Subject(s)
Microplastics , Soil , Microplastics/analysis , Plastics/analysis , Sodium Chloride , ChinaABSTRACT
INTRODUCTION: This study aimed to understand the transcriptome characteristics of the skeletal muscle of elderly (EL) men with metabolic syndrome (MS) and to find the hub genes and insight into the molecular mechanisms of skeletal muscle in the occurrence and development of MS. METHODS: In this study, the limma package of R software was used to analyze the differentially expressed genes in the skeletal muscle of healthy young (YO) adult men, healthy EL men, and EL men diagnosed with MS (SX) for at least 10 years. Bioinformatics methods, such as GO enrichment analysis, KEGG enrichment analysis, and gene interaction network analysis, were used to explore the biological functions of differentially expressed genes, and weighted gene coexpression network analysis (WGCNA) was used to cluster differentially expressed genes into modules. RESULTS: Among the YO group, EL group, and SX group, 65 co-differentially expressed genes were found maybe regulated by age factor and MS factor. Those co-differentially expressed genes were enriched into 25 biological process terms and 3 KEGG pathways. Based on the WGCNA results, a total of five modules were identified. Fifteen hub genes may play an essential role in regulating the function of skeletal muscle of EL men with MS. CONCLUSIONS: 65 differentially expressed genes and 5 modules may regulate the function of skeletal muscle of EL men with MS, among which fifteen hub genes may play an essential role in the occurrence and development of MS.
Subject(s)
Metabolic Syndrome , Transcriptome , Male , Humans , Aged , Metabolic Syndrome/genetics , Gene Expression Profiling , Gene Regulatory Networks , Muscle, Skeletal/metabolismABSTRACT
Cypermethrin (CP) exhibits anti-androgenic effects through antagonism on androgen receptor (AR) activation. This study was to identify whether AR-mediated disabled 2 interacting protein (DAB2IP)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway was involved in CP-induced mouse Sertoli cells (TM4) proliferation disorder. Real-Time Cell Analysis-iCELLigence system was to measure cell proliferation. Bioinformatic analyses were performed to identify AR-regulated genes. Quantitative Real-Time PCR and western blot were to detect the genes and proteins levels in AR-mediated DAB2IP/PI3K/AKT pathway. Results showed CP suppressed TM4 proliferation and the expression of AR. Activation of AR restored the inhibition efficacy of CP on TM4 proliferation. AR regulated DAB2IP expression and phosphorylation levels of PI3K and AKT in CP-exposed TM4 cells. In addition, knockdown of DAB2IP alleviated the inhibition efficacy of CP on cell proliferation and phosphorylation of PI3K and AKT. Taken together, AR was a modulator in CP-induced inhibition of Sertoli cells proliferation by negatively regulating DAB2IP/PI3K/AKT signaling pathway. The study may provide a new insight for the mechanisms of male reproductive toxicity induced by CP.
ABSTRACT
Cypermethrin, an extensively used pyrethroid pesticide, is regarded as one of many endocrine-disrupting chemicals (EDCs) with anti-androgenic activity to damage male reproductive systems. We previously found cypermethrin-induced apoptosis in mouse Sertoli cells TM4. We hypothesized cypermethrin-induced TM4 apoptosis by the endoplasmic reticulum (ER) pathway. This study aimed to explore the roles of the ER pathway in cypermethrin-induced apoptosis in TM4 cells. The cells were treated with cypermethrin for 24 h at various concentrations (0 µM, 10 µM, 20 µM, 40 µM, and 80 µM). Flow cytometry was used to test for apoptosis. Western blot was used to test protein expressions in the ER stress pathway. The results showed that the apoptosis rate of TM4 cells increased with increased concentrations of cypermethrin, and a significant difference was detected in the 80-µM group. The protein expressions of glucose-regulated protein 78 (GRP78), protein kinase R (PKR)-like ER kinase (PERK), p-PERK, α subunit of eukaryotic initiation factor (eIF2α), p-eIF2α, activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), caspase-12, caspase-9, and caspase-3 increased with increased concentrations of cypermethrin. The results suggested cypermethrin-induced apoptosis in TM4 cells regulated by the ER pathway involving PERK-eIF2α-ATF4-CHOP. The study provides a new insight into cypermethrin-induced apoptosis in Sertoli cells.
Subject(s)
Pyrethrins , eIF-2 Kinase , Activating Transcription Factor 4/metabolism , Animals , Apoptosis , Endoplasmic Reticulum , Eukaryotic Initiation Factor-2/metabolism , Male , Mice , Pyrethrins/toxicity , Sertoli Cells , Signal Transduction , eIF-2 Kinase/metabolismABSTRACT
With the rapid increase in the use of plastic films, microplastic (MP) pollution in agricultural soils has become a global environmental problem. Propiconazole is widely used in agriculture and horticulture; however, its role in plastic film degradation remains elusive. Butylene adipate-co-terephthalate (PBAT) and polyethylene (PE) films were used to analyze the effects of propiconazole on plastic film and MP degradation. We identified the surface morphologies of PBAT and PE at different propiconazole concentrations and soil pH values, as well as the adsorption and release characteristics of heavy metals during the degradation process via scanning electron microscopy, Fourier transform infrared spectroscopy and inductively coupled plasma mass spectrometry. Propiconazole accelerated the degradation of MPs, adsorption of heavy metals (Ni and Zn), and release of Sn at low concentrations (≤40 mg/kg); however, these effects were evidently absent at a high concentration (120 mg/kg). Furthermore, MPs were more prone to degradation in acidic or alkaline soils than in neutral soil when they coexisted with propiconazole. Hence, we suggest that PBAT and PE plastic films may not be suitable for application in acidic and alkaline soils with propiconazole, because of shorter rupture time and more heavy metal adsorption. PBAT degraded faster, absorbed and released more heavy metals than PE. Under all tested conditions, the heavy metal contents in MPs gradually approached those in soil, which proves that MPs are carriers of heavy metal pollutants. These results may help in assessing the impact of MPs on soil environments and provide a theoretical basis for the standardized propiconazole and plastic film usage.