1. [Bisphenol A alters glucose metabolism in rat Sertoli cells in vitro].
- Author
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Huang W, Huang HM, Wang H, Zhao JC, Li MZ, Wang HQ, Wang XS, and Wang PT
- Subjects
- Animals, Benzhydryl Compounds administration & dosage, Cell Proliferation drug effects, Cells, Cultured, Culture Media, Serum-Free, Dimethyl Sulfoxide pharmacology, In Vitro Techniques, Infertility, Male chemically induced, Male, Phenols administration & dosage, RNA, Messenger metabolism, Random Allocation, Rats, Rats, Wistar, Sertoli Cells metabolism, Spermatogenesis drug effects, Benzhydryl Compounds pharmacology, Glucose metabolism, L-Lactate Dehydrogenase metabolism, Phenols pharmacology, Sertoli Cells drug effects
- Abstract
Objective: To observe the influence of different concentrations of bisphenol A (BPA) on glucose metabolism and lactate dehydrogenase (LDH) expression in rat Sertoli cells in vitro and investigate the mechanisms of BPA inducing male infertility., Methods: Using two-step enzyme digestion, we isolated Sertoli cells from male Wistar rats and constructed a primary Sertoli cell system, followed by immunohistochemical FasL staining. We randomly divided the Sertoli cells into a control group to be cultured in the serum-free minimal essential medium (MEM) plus dimethyl sulfoxide (DMSO) and three experimental groups to be treated with 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA, respectively, in the MEM plus DMSO. After 48 hours of treatment, we measured the proliferation of the cells by CCK-8 assay, determined the concentrations of metabolites by NMR spectroscopy, and detected the expression of LDH in the Sertoli cells by RT-PCR and Western blot., Results: The purity of the isolated Sertoli cells was (96.05 ± 1.28)% (n = 10). Compared with the control group, the 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA groups showed no remarkable changes in the proliferation of Sertoli cells ([98 ± 8]%, [96 ± 3]%, and [95 ± 3]%, P >0.05), but the 10 μmol/L and 1 mmol/L of BPA groups exhibited significantly decreased concentrations of intracellular glucose ([3.89 ± 0.07] vs [3.36 ± 0.24] and [3.04 ± 0.21] pmol/cell, P <0.05) and lactate ([0.43 ± 0.06] vs [0.29 ± 0.05] and [0.20 ± 0.03] pmol/cell, P <0.05). The expression of LDH mRNA was decreased with the increased concentration of BPA, while that of LDH protein reduced only in the 1 mmol/L BPA group (P <0.05)., Conclusion: High-concentration BPA decreases the expression of LDH and alters glucose metabolism in Sertoli cells, and therefore may reduce the provision of lactate for germ cells and impair spermatogenesis.
- Published
- 2015