NF-κB Pathway Contributes to Cadmium-Induced Apoptosis of Porcine Granulosa Cells

• Published in: Biological trace element research 2013-06, Vol.153 (1-3), p.403-410
• Main Authors: Wang, Chunqiang, Ma, Wei, Su, Yuhong
• Format: Article
• Language: English
• Subjects: acridine orange; Animals; apoptosis; Apoptosis - drug effects; Biochemistry; Biomedical and Life Sciences; Biotechnology; cadmium; Cadmium - toxicity; ethidium; Female; fluorescent antibody technique; genes; granulosa cells; Granulosa Cells - cytology; Granulosa Cells - drug effects; Granulosa Cells - metabolism; Life Sciences; NF-kappa B - metabolism; Nutrition; Oncology; oxidation; proteins; reactive oxygen species; Reactive Oxygen Species - metabolism; Sesquiterpenes - pharmacology; Swine; transcription factor NF-kappa B; Western blotting
• ISSN: 0163-4984; 1559-0720
• DOI: 10.1007/s12011-013-9650-7

To better understand the mechanism of cadmium (Cd)-induced apoptosis of porcine granulosa cells, we examined the nuclear factor-kappa B (NF-κB) p65 subunits intracellular translocation and the expression of some downstream apoptotic-related genes. Apoptosis and reactive oxygen species (ROS) production in porcine granulosa cells exposed to cadmium chloride (CdCl₂) were determined by acridine orange/ethidium bromide double staining and 2,7-dichlorodihydro-fluorescein-diacetate oxidation staining, respectively. The results showed that the apoptosis of porcine granulosa cells induced by CdCl₂ significantly increased in a time- and dose-dependent manner along with the increasing of ROS production, and 10 μM parthenolide, an inhibitor NF-κB, can accelerate the process of apoptosis. Moreover, immunofluorescence and western blot results showed that CdCl₂ could stimulate the translocation of p65 into nucleus in porcine granulosa cells. Furthermore, CdCl₂ also significantly stimulate the expression of Bcl-2 proteins in porcine granulosa cells than that in the control. In contrast, we did not find any change of Bax expression in granulosa cells upon exposure of cadmium. Taken together, these results demonstrate that the activation of NF-κB pathway may play a crucial role in cadmium-induced apoptosis of porcine granulosa cells.