Oxidative stress, DNA damage, and cellular response in hydrogen peroxide-induced cell injury of mud crab (Scylla paramamosain)

doi:10.1016/j.fsi.2021.04.015

Fish & Shellfish Immunology

Volume 114, July 2021, Pages 82-89

https://doi.org/10.1016/j.fsi.2021.04.015 Get rights and content

Highlights

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First transcriptome of the mud crab response to hydrogen peroxide stress.
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H₂O₂ activated FOXO signaling pathway.
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DNA damage response and apoptosis signaling pathways played important roles in response to oxidative stress.

Abstract

Oxidative stress is considered as the toxicity mechanism of environmental stressors on aquatic organisms. This study aims to explore the effects of oxidative stress on physiological responses, DNA damage and transcriptional profiles of the mud crabs Scylla paramamosain. In the present study, mud crabs were injected with 0.1% and 1% hydrogen peroxide (H₂O₂) for 72 h. The results showed that superoxide dismutase and catalase activities significantly decreased after H₂O₂ injection. Malondialdehyde content, H₂O₂ content, aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase activity significantly increased after H₂O₂ injection. Moreover, DNA damage occurred after H₂O₂ injection. Transcriptome analysis showed that 531 and 372 differentially expressed genes (DEGs) were identified after 0.1% and 1% H₂O₂ injection, respectively. These DEGs were mainly involved in the oxidative stress response and immune functions. All these results indicated that oxidative stress could impair both antioxidant defense systems and immune systems. Transcriptome analysis provided valuable information on gene functions associated with the response to oxidative stress in the mud crab.

Introduction

The mud crab (Scylla paramamosain) is one of the most important marine crustacean species in China. Its production exceeded 158,000 tons in 2018. However, mud crab farming suffers from large economic losses due to disease outbreaks. Mud crabs are subjected to various environmental stressors, causing a depression in the immune response and an increased vulnerability to disease [1]. A previous study has shown that ammonia exposure disturbed physiological responses, and caused liver damage in the mud crab [2]. Oxidative stress is considered to be one of the toxicity mechanisms of environmental stressors on aquatic organisms [3]. This results from an imbalance between the antioxidant system, and reactive oxygen species (ROS) production. ROS includes: superoxide anion, hydrogen peroxide and hydroxyl radicals. Under oxidative stress, the overproduction of ROS can damage important biomolecules or result in cell death. In contrast, organisms evolve some strategies to protect cells against the oxidative damage. Thus, understanding the defense mechanism against oxidative stress is important for mud crab farming.

Hydrogen peroxide (H₂O₂), a major ROS, is a strong oxidizing agent that can oxidize and damage proteins, lipids, and DNA, which causes oxidative damage [4]. H₂O₂ was used as a model chemical to study the oxidative stress in vivo and in vitro [5,6]. In aquaculture, H₂O₂ is used as a disinfectant due to its relatively lower toxicity [7]. H₂O₂ is also used to control bacteria and parasites in fish [8]. Excessive H₂O₂ in the aquatic environment can easily pass through cell membranes, and cause oxidative stress [9]. Many studies have reported the effects of H₂O₂ toxicity on aquatic animals. Roque et al. [10] suggested that stress indicators, such as plasma cortisol and hemoglobin levels, were changed in Dicentrarchus labrax after H₂O₂ exposure. H₂O₂ exposure decreased the innate immune response in Paralichthys olivaeceus [11]. H₂O₂ treatment caused cellular inflammation and severe hepatotoxic damage in Oreochromis niloticus [12]. However, the mechanism involved in H₂O₂-induced oxidative damage in crustaceans is still poorly understood.

In the present study, we used H₂O₂ to study the effects of oxidative stress on physiological responses, DNA damage, and the transcriptional profiles of mud crabs. Our study provides a useful foundation for understanding the mechanisms of oxidative stress in crustaceans.

Section snippets

Animals

Mud crabs (50 ± 3.6 g) were purchased in Taishan city (Guangdong Province, China). Before the experiment, the mud crabs were acclimated in 200 L tanks with of 10‰ salinity seawater at 25–27 °C for one week. They were fed oyster meat twice a day.

Mud crabs were assigned at random to three experimental groups. Each treatment group consisted of 60 mud crabs. There were three replicates per treatment. Through preliminary experiments, we found that 96 h LC50 was approximately 5% the value of H₂O₂. In

Effect of H₂O₂ on oxidative stress

Compared with the control group, the H₂O₂ content in the hepatopancreas significantly increased from 6 to 72 h after the 0.1% and 1% H₂O₂ injection (Fig. 1A). SOD and CAT activity did not change at 6 h after the 0.1% H₂O₂ injections. SOD and CAT activity significantly decreased from 12 to 72 h after 0.1% and 1% H₂O₂ injection (Fig. 1B and C). Compared with the control group, the MDA content was higher values from 6 to 72 h after the 0.1% and 1% H₂O₂ injections (Fig. 1D).

Effect of H₂O₂ on biochemical parameters

As shown in Fig. 2A, ALT

Discussion

Environmental stress is one of the most serious problems in aquaculture because it can cause a depression in the immune ability and even death. Oxidative stress is considered a mechanism of toxicity in aquatic organisms under environmental stress. In the present study, H₂O₂ was used to induce oxidative stress. This study provides evidence of the mechanisms behind the response to oxidative stress in mud crabs.

H₂O₂ can readily diffuse across cell membranes. In this study, H₂O₂ content in the

CRediT authorship contribution statement

Chang-Hong Cheng: Conceptualization, Methodology, Formal analysis, Investigation, Writing – original draft. Hong-Ling Ma: Resources. Guang-Xin Liu: Supervision. Yi-Qin Deng: Methodology, Supervision. Juan Feng: Writing – review & editing. Yu-Kun Jie: Methodology, Formal analysis. Zhi-Xun Guo: Funding acquisition, Writing – review & editing.

Acknowledgments

This research was supported by the China Agriculture Research System (CARS-48), National Natural Science Foundation of China (32002380), Central Public-interest Scientific Institution Basal Research Fund, CAFS (2020TD42 and 2021SD05), Basic and applied basic research fund of Guangdong Province (2019A1515011548 and 2021A1515012483), Science and Technology Program Project of Guangzhou (201904010327), Central Public-interest Scientific Institution Basal Research Fund, South China Sea Fisheries

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Full length articleOxidative stress, DNA damage, and cellular response in hydrogen peroxide-induced cell injury of mud crab (Scylla paramamosain)

Highlights

Abstract

Introduction

Section snippets

Animals

Effect of H2O2 on oxidative stress

Effect of H2O2 on biochemical parameters

Discussion

CRediT authorship contribution statement

Acknowledgments

Ecotoxicol. Environ. Saf.

Environ. Toxicol. Pharmacol.

Fish Shellfish Immunol.

Aquaculture

Aquaculture

Aquat. Toxicol.

Anal. Biochem.

Aquat. Toxicol.

Anal. Biochem.

Methods

Chemosphere

Aquat. Toxicol.

J. Exp. Mar. Biol. Ecol.

Chemosphere

Rev Adv Canc Res

Agricult Natural Resour

Chemosphere

Fish Shellfish Immunol.

J. Inorg. Biochem.

Full length article
Oxidative stress, DNA damage, and cellular response in hydrogen peroxide-induced cell injury of mud crab (Scylla paramamosain)

Effect of H₂O₂ on oxidative stress

Effect of H₂O₂ on biochemical parameters