Zinc-pretreatment triggers glutathione and Nrf2-mediated protection against inorganic mercury-induced cytotoxicity and intrinsic apoptosis in PC12 cells

Highlights

• Inorganic Hg induced intrinsic apoptosis in PC12 cells via ROS generation.

• Zinc (Zn)-pretreatment inhibited inorganic Hg induced toxicity and apoptosis.

• Zn-protection in iHg mediated toxicity is via its antioxidant properties.

• Zn improved glutathione content and ROS amount against Hg-induced toxicity.

• Zn upregulated nrf2 and HO1 in Hg induced cytotoxicity.

Abstract

Mercury (Hg) is a hazardous metal, poses environmental problems with severe human health effects; whereas zinc (Zn) is an essential micronutrient with antioxidant properties. The purpose of this research was to investigate the effect of Zn on inorganic Hg-induced cytotoxicity in the PC12 cells. The cells were treated with HgCl₂ (5 μM) for 48 h with/without 1 h prior ZnCl₂-treatment (100 μM) and deliberated for further analysis. After 48 h of incubation with only Hg²⁺, the cell showed reduced cell viability, compromised cell membrane, DNA degradation, depleted glutathione level, ROS generation and drastically increased apoptosis. Subsequently, Hg²⁺-treated cells demonstrated a significant downregulation of akt, mTOR, ERK1, Nrf2, HO1, Bcl-2, Bcl-xL, and upregulation of p53, Bax, cytochrome c and cleaved caspase 3, indicating intrinsic apoptosis induction. However, cells pretreated with Zn²⁺ before Hg²⁺-exposure showed a significant improvement in cell viability, cell membrane, DNA damage, glutathione level, ROS amount and apoptotic cells, with a significant upregulation in mTOR, akt, ERK1, Nrf2, HO1, Bcl-2 and Bcl-xL, and downregulation in p53, Bax, cytochrome c and cleaved caspase 3, indicating inhibition of apoptosis. The findings suggested that Zn²⁺-pretreatment not only improves glutathione content but also induces activation of Nrf2-HO1 pathway, which would tend to suppress Hg-cytotoxicity.