Sulforaphane Protect Against Cadmium-Induced Oxidative Damage in mouse Leydigs Cells by Activating Nrf2/ARE Signaling Pathway

• Published in: International journal of molecular sciences 2019-02, Vol.20 (3), p.630
• Main Authors: Yang, Shu-Hua, Li, Peng, Yu, Li-Hui, Li, Lin, Long, Miao, Liu, Ming-Da, He, Jian-Bin
• Format: Article
• Language: English
• Subjects: Animals; Antioxidant Response Elements - drug effects; Antioxidants; Antioxidants - pharmacology; Apoptosis; Apoptosis - drug effects; Arsenic; Ascorbic acid; Atrophy; Cadmium; Cadmium Chloride - antagonists & inhibitors; Cadmium Chloride - toxicity; Cell Line; Cell Survival - drug effects; Data analysis; Diabetes mellitus; Dose-Response Relationship, Drug; Enzymes; Flow cytometry; Gene Expression Regulation; Glutamate-Cysteine Ligase - genetics; Glutamate-Cysteine Ligase - metabolism; Glutathione Peroxidase - genetics; Glutathione Peroxidase - metabolism; Heme Oxygenase-1 - genetics; Heme Oxygenase-1 - metabolism; Isothiocyanates - pharmacology; Leydig Cells - cytology; Leydig Cells - drug effects; Leydig Cells - metabolism; Male; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; NAD(P)H Dehydrogenase (Quinone) - genetics; NAD(P)H Dehydrogenase (Quinone) - metabolism; NF-E2-Related Factor 2 - agonists; NF-E2-Related Factor 2 - genetics; NF-E2-Related Factor 2 - metabolism; Nrf2/ARE signaling pathway; oxidative damage; Oxidative Stress - drug effects; Protein expression; Reactive Oxygen Species - antagonists & inhibitors; Reactive Oxygen Species - metabolism; Reproductive disorders; SFN; Signal transduction; Signal Transduction - drug effects; Signal Transduction - genetics; Sperm; Sulforaphane; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Testosterone; Testosterone - biosynthesis; TM3 cell; Toxicity; Tubules; Weight reduction
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20030630

Cadmium (Cd) is harmful for humans and animals, especially for the reproductive system. However, the mechanism of its toxicity has not been elucidated, and how to alleviate its toxicity is very important. This study aimed to explore the role and mechanism of action of sulforaphane (SFN) in protecting mouse Leydigs (TM3) cells from cadmium (Cd)-induced damage. The half-maximal inhibitory concentration (IC ) of Cd and the safe doses of SFN were determined using a methyl thiazolyl tetrazolium (MTT) assay. The testosterone secretion from TM3 cells was measured using the enzyme-linked immunosorbent assay. The intracellular oxidative stress was evaluated using corresponding kits. The cell apoptosis was detected using flow cytometry. The mRNA expression of genes associated with NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling was detected using reverse transcription⁻polymerase chain reaction, including Nrf2, heme oxygenase I (HO-1), glutathione peroxidase (GSH-Px), NAD(P)H:quinone acceptor oxidoreductase 1 (NQO1), and γ-glutamylcysteine synthetase (γ-GCS). The protein expression of Nrf2, GSH-Px, HO-1, γ-GCS, and NQO1 was detected using Western blot analysis. The results showed that the IC of Cd to TM3 cells was 51.4 µmol/L. SFN reduced the release of lactate dehydrogenase from Cd-exposed cells. Cd + SFN 2.5 treatment significantly elevated testosterone concentration compared with the Cd group ( < 0.05). SFN significantly increased total superoxide dismutase (T-SOD) and GSH-Px activity and GSH content in Cd-treated cells ( < 0.05; < 0.01), inhibited the production of malondialdehyde or reactive oxygen species caused by Cd ( < 0.05; < 0.01), and reduced the apoptotic rate of Cd-induced TM3 cells ( < 0.01). SFN upregulated the mRNA expression of , , , , and in Cd-treated cells, indicating the protective effect of SFN against Cd-induced oxidative stress or cell apoptosis by activating the Nrf2/ARE signaling pathway.