GC-1 spg

DEP Induces Oxidative Stress in GC-1 spg Cells

DEP particles trigger oxidative stress and inflammation which negatively affects male reproductive health by disrupting spermatogenesis. Shi's team investigates how DEP triggers oxidative stress in GC-1 spg cells by analyzing the role of reactive oxygen species (ROS) buildup and the Keap1-Nrf2 signaling pathway.

They investigated oxidative stress contributions to DEP-induced cytotoxic effects on GC-1 spg cells by measuring cellular oxidative damage through biochemical indices. After DEP exposure, DCFH-DA fluorescence intensity increased significantly in a dose-dependent manner, showing that DEP increased ROS formation in GC-1 spg cells (Fig. 1a and b). Excess ROS can damage cell membranes and cause mitochondrial injury. We assessed mitochondrial membrane potential (MMP) using JC-1 dye. As DEP concentration increased, J-monomer fluorescence increased, while J-aggregate fluorescence decreased. Quantitative analysis showed a significant decrease in the J-aggregate/J-monomer ratio in DEP-treated cells, indicating mitochondrial depolarization (Fig. 1c and d). MDA is a marker for lipid peroxidation. In GC-1 spg cells treated with DEP, MDA concentration increased with DEP concentration (Fig. 1e). They also measured GSH levels, which decreased with DEP treatment (Fig. 1f). SOD activity was also reduced in DEP-exposed cells. The results show that DEP treatment caused oxidative damage and stimulated antioxidant defenses.

Fig. 1. DEP induced oxidative stress and mitochondrial damage in GC-1spg cell (Shi J, Tian F, et al., 2024).

ZnO NPsInduces Apoptosis and Autophagy of Mouse GC-1 spg Cells

Zinc oxide nanoparticles (ZnO NPs), widely used for their unique properties, have raised health concerns due to their potential to induce reproductive toxicity, particularly affecting spermatogenesis. Yang's team investigated how oxidative stress contributes to ZnO NPs-induced apoptosis and autophagy in mouse GC-1 spg cells, shedding light on mechanisms underlying reproductive toxicity and potential spermatogenesis failure.

ZnO NPs inhibited GC-1 spg cell viability in a dose-dependent manner (Fig. 2A). They increased protein levels of cleaved Caspase-8, Caspase-3, and Bax, while reducing Bcl-2 levels (Fig. 2B and C). ZnO NPs also raised early (AnnexinV+/PI?) and late apoptosis (AnnexinV+/PI+) cell counts (Fig. 2D and E), indicating apoptosis induction. ZnO NPs significantly raised LC3 content, LC3-II/LC3-I ratio, and ATG 5, Beclin 1 protein levels (Fig. 3A and B). TEM revealed few autophagosomes in control cells but many in ZnO NPs- and starvation-treated cells, which showed organelle degradation like mitochondria and endoplasmic reticulum (Fig. 3C), indicating autophagy induction.

Fig. 2. ZnO NPs induces apoptosis of mouse GC-1 spg cells (Yang D, Zhang M, et al., 2020).

Fig. 3. ZnO NPs induces autophagy of mouse GC-1 spg cells (Yang D, Zhang M, et al., 2020).