Journal: Reproductive toxicology (Elmsford, N.Y.)

Article Title: 3-chloro-1,2-propanediol induces oxidative stress and promotes testicular damage and infertility in rats through CYP2C9.

doi: 10.1016/j.reprotox.2024.108633

Figure Lengend Snippet: Fig. 6. 3-MCPD affects oxidative stress and promotes testicular cell apoptosis through CYP2C9. (A) The 2D structure of 3-MCPD. (B) Visualization result of molecular docking. (C) Western blot analysis of the CYP2C9 in R2C cells under control, low dose 3-MCPD (0.5 mmol/L), or high dose (2 mmol/L), (D) Western blot analysis of the CYP2C9 in R2C cells under control, high dose 3-MCPD (2 mmol/L) or 3-MCPD + sulphametoxazole (CYP2C9 inhibitor, SPZ) treatments for 24 hours. (E) Proliferation detection by CCK8 assay. (F) Progesterone synthesized quantitatively measured. (G) ROS levels using ELISA assay against R2C cell line. (H) Oxidative stress-related molecules HO-1 and SOD were detected by Western blot. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: R2C rat Leydig tumor cells (catalog no. CCL-97) was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Western Blot, Control, CCK-8 Assay, Synthesized, Enzyme-linked Immunosorbent Assay

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: (A) Representative aspect of cells after 24 h of exposure to ethylene dimethanesulfonate (EDS) of rat R2C and mouse MA-10 Leydig cells. Note that at 5 mM, the cell confluency and morphological characteristic seem qualitatively impaired (red arrows; insert), indicating the cytotoxicity caused by EDS. (B) Cell count of R2C and MA-10 cells after 4 and 24 h of EDS exposure at concentrations of 1 or 2 mM (n = 4, each in duplicate). Values are expressed as median (interquartile ranges). Kruskal-Wallis followed by Dunn’s test (p > 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Cell Counting

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: A dose–response study with reporter luciferase assays to determine the effects of EDS on (A) Star , (B) Cyp17a1 , (C) Insl3 , and (D) Gsta3 promoter activity. Rat R2C (n = 4, each in triplicate) and mouse MA-10 (n = 5, each in triplicate) Leydig cells were transfected with different promoter constructs and treated for 24 h with increasing concentrations of EDS as indicated. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. ANOVA followed by Dunnett’s test (*p < 0.05 compared to the control group).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Luciferase, Activity Assay, Transfection, Construct, Control

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: EDS effects on the activity of the Insl3 and Gsta3 gene promoter after 4 and 24 h of exposure to EDS of rat R2C (1 mM of EDS, n = 4–5, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Activity Assay, Luciferase

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: EDS effects on Star promoter activity after 4 h of exposure to EDS of rat R2C (1 mM of EDS, n = 5, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells. EDS effects on Star promoter activity after 24 h of exposure to EDS of rat R2C (1 mM of EDS, n = 3, each in triplicate) and mouse MA-10 (2 mM of EDS, n = 5, each in triplicate) Leydig cells, DC3 granulosa cells (1 mM of EDS, n = 5, each in triplicate), and MSC-1 Sertoli cells (2 mM of EDS, n = 5, each in triplicate). EDS effects were assessed in basal and stimulated (0.5 mM 8Br-cAMP) conditions in R2C Leydig cells. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Activity Assay, Luciferase

Journal: Current Research in Toxicology

Article Title: Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

doi: 10.1016/j.crtox.2023.100147

Figure Lengend Snippet: The EDS-responsive region is located between −400 and −195 bp of the Star promoter. Progressive 5′ deletion constructs of Star gene promoter were transfected in rat R2C Leydig cells and treated with 1 mM EDS for 24 h (n = 3–5, each in triplicate) in the absence or presence of 0.5 mM 8Br-cAMP for 4 h. Luciferase reporter (Luc). Values are expressed as mean ± S.E.M. Welch’s t -test (*p < 0.05).

Article Snippet: Rat and mouse immortalized Leydig cell lines, R2C (ATCC, Manassas, Virginia, USA) and MA-10 (ATCC, Manassas, Virginia, USA), respectively, were used for this study from passages 40 to 52 (R2C) or 08 to 31 (MA-10).

Techniques: Construct, Transfection, Luciferase