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Microcystin-LR Induced Apoptosis in Rat Sertoli Cells via the Mitochondrial Caspase-Dependent Pathway: Role of Reactive Oxygen Species

View Article: PubMed Central - PubMed

ABSTRACT

Microcystins (MCs), the secondary metabolites of blue-green algae, are ubiquitous and major cyanotoxin contaminants. Besides the hepatopancreas/liver, the reproductive system is regarded as the most important target organ for MCs. Although reactive oxygen species (ROS) have been implicated in MCs-induced reproductive toxicity, the role of MCs in this pathway remains unclear. In the present study, Sertoli cells were employed to investigate apoptotic death involved in male reproductive toxicity of microcystin-LR (MC-LR). After exposure to various concentrations of MC-LR for 24 h, the growth of Sertoli cells was concentration-dependently decreased with an IC50 of ~32 μg/mL. Mitochondria-mediated apoptotic changes were observed in Sertoli cells exposed to 8, 16, and 32 μg/mL MC-LR including the increased expression of caspase pathway proteins, collapse of mitochondrial membrane potential (MMP), and generation of ROS. Pretreatment with a global caspase inhibitor was found to depress the activation of caspases, and eventually increased the survival rate of Sertoli cells, implying that the mitochondrial caspases pathway is involved in MC-LR-induced apoptosis. Furthermore, N-acetyl-l-cysteine attenuated the MC-LR-induced intracellular ROS generation, MMP collapse and cytochrome c release, resulting in the inhibition of apoptosis. Taken together, the observed results suggested that MC-LR induced apoptotic death of Sertoli cells by the activation of mitochondrial caspases cascade, while its effects on the ROS-mediated signaling pathway may contribute toward the initiation of mitochondrial dysfunction.

No MeSH data available.


Related in: MedlinePlus

Effect of MC-LR on the proliferation inhibition of rat Sertoli cells using CCK-8 assay. Cells were exposed to MC-LR at different concentrations (0, 1, 5, 10, 20, 40, and 60 μg/mL) for 24 h, and the optical density (OD) was detected with CCK-8 assay. Data were presented as mean ± SEM of triplicate independent experiments. *P < 0.05 vs. control group.
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Figure 1: Effect of MC-LR on the proliferation inhibition of rat Sertoli cells using CCK-8 assay. Cells were exposed to MC-LR at different concentrations (0, 1, 5, 10, 20, 40, and 60 μg/mL) for 24 h, and the optical density (OD) was detected with CCK-8 assay. Data were presented as mean ± SEM of triplicate independent experiments. *P < 0.05 vs. control group.

Mentions: To evaluate the effect of MC-LR on the growth of Sertoli cells, we treated the cells with a gradient of concentration of MC-LR for 24 h. The results indicated that the cell growth was inhibited in a concentration-dependent manner (Figure 1), and the IC50 dose of MC-LR for 24 h was determined to be 32 μg/mL. Thus, 8, 16, and 32 μg/mL of MC-LR (IC50/4, IC50/2, and IC50, respectively) were used in subsequent experiments.


Microcystin-LR Induced Apoptosis in Rat Sertoli Cells via the Mitochondrial Caspase-Dependent Pathway: Role of Reactive Oxygen Species
Effect of MC-LR on the proliferation inhibition of rat Sertoli cells using CCK-8 assay. Cells were exposed to MC-LR at different concentrations (0, 1, 5, 10, 20, 40, and 60 μg/mL) for 24 h, and the optical density (OD) was detected with CCK-8 assay. Data were presented as mean ± SEM of triplicate independent experiments. *P < 0.05 vs. control group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5016609&req=5

Figure 1: Effect of MC-LR on the proliferation inhibition of rat Sertoli cells using CCK-8 assay. Cells were exposed to MC-LR at different concentrations (0, 1, 5, 10, 20, 40, and 60 μg/mL) for 24 h, and the optical density (OD) was detected with CCK-8 assay. Data were presented as mean ± SEM of triplicate independent experiments. *P < 0.05 vs. control group.
Mentions: To evaluate the effect of MC-LR on the growth of Sertoli cells, we treated the cells with a gradient of concentration of MC-LR for 24 h. The results indicated that the cell growth was inhibited in a concentration-dependent manner (Figure 1), and the IC50 dose of MC-LR for 24 h was determined to be 32 μg/mL. Thus, 8, 16, and 32 μg/mL of MC-LR (IC50/4, IC50/2, and IC50, respectively) were used in subsequent experiments.

View Article: PubMed Central - PubMed

ABSTRACT

Microcystins (MCs), the secondary metabolites of blue-green algae, are ubiquitous and major cyanotoxin contaminants. Besides the hepatopancreas/liver, the reproductive system is regarded as the most important target organ for MCs. Although reactive oxygen species (ROS) have been implicated in MCs-induced reproductive toxicity, the role of MCs in this pathway remains unclear. In the present study, Sertoli cells were employed to investigate apoptotic death involved in male reproductive toxicity of microcystin-LR (MC-LR). After exposure to various concentrations of MC-LR for 24 h, the growth of Sertoli cells was concentration-dependently decreased with an IC50 of ~32 &mu;g/mL. Mitochondria-mediated apoptotic changes were observed in Sertoli cells exposed to 8, 16, and 32 &mu;g/mL MC-LR including the increased expression of caspase pathway proteins, collapse of mitochondrial membrane potential (MMP), and generation of ROS. Pretreatment with a global caspase inhibitor was found to depress the activation of caspases, and eventually increased the survival rate of Sertoli cells, implying that the mitochondrial caspases pathway is involved in MC-LR-induced apoptosis. Furthermore, N-acetyl-l-cysteine attenuated the MC-LR-induced intracellular ROS generation, MMP collapse and cytochrome c release, resulting in the inhibition of apoptosis. Taken together, the observed results suggested that MC-LR induced apoptotic death of Sertoli cells by the activation of mitochondrial caspases cascade, while its effects on the ROS-mediated signaling pathway may contribute toward the initiation of mitochondrial dysfunction.

No MeSH data available.


Related in: MedlinePlus