Cyclosporine A Induces Cardiac Developmental Toxicity in Zebrafish by Up-Regulation of Wnt Signaling and Oxidative Stress

Abstract

Due to the widely application of Cyclosporine A (CsA) as an immunosuppressant in clinic, it is necessary to study its potential toxicity. Therefore, we used zebrafish as a model animal to evaluate the toxicity of CsA on embryonic development. Exposure of zebrafish embryos to CsA at concentrations of 5 mg/L, 10 mg/L, and 15 mg/L from 12 hpf to 72 hpf resulted in abnormal embryonic development, including cardiac malformation, pericardial edema, decreased heart rate, decreased blood flow velocity, deposition at yolk sac, shortened body length, and increased distance between venous sinus and arterial bulb (SV-BA). The expression of genes related to cardiac development was disordered, and the apoptotic genes were up-regulated. Oxidative stress level was up-regulated and accumulated in pericardium in a dose-dependent manner. Astaxanthin (ATX) treatment could significantly alleviate zebrafish heart defects. CsA induced up-regulation of Wnt signaling in zebrafish, and IWR-1, an inhibitor of Wnt signaling pathway, could effectively rescue the heart defects induced by CsA. Together, our study indicated that CsA induced cardiac developmental toxicity in zebrafish larvae through up-regulating oxidative stress and Wnt signaling, contributing to a more comprehensive evaluation of the safety of the drug.

Pentachloronitrobenzene Reduces the Proliferative Capacity of Zebrafish Embryonic Cardiomyocytes via Oxidative Stress

“…This finding was consistent with the previous reports that the reduced levels of the tbx2b gene inhibited cardiac proliferation [22]. We also performed H&E staining of the paraffin sections of the heart and found that the heart was significantly linearized, whereas the number of cardiomyocytes was significantly reduced [19]. This observation suggested that the abnormal zebrafish heart development caused by PCNB might be due to the decreased proliferation of heart cells and substantial reduction in the ability of the heart to loop.…”

“…All zebrafish breeds were obtained from the National Zebrafish Resource Center and were raised under standard temperature (28 ± 1 • C) and 14 h light:10 h dark photoperiod conditions according to the Institutional Animal Care and Use Committee. Breeding zebrafish requires water with a pH of 7 and a conductivity of 500 µS/cm, feeding at 9:00 a.m. and 2:00 p.m. every day with brine shrimp larvae [19]. All zebrafish embryos were euthanized by ice bath after experimentation.…”

“…As an ideal animal model for cardiovascular research [17], zebrafish has been widely used in the screening of drug cardiotoxicity. For example, in 2020, Huang et al, studied the cardiotoxicity of the herbicide Oxadiazon-Butachlor [18] on zebrafish, and in 2021, Wan et al, analyzed the toxicity of the clinical drug Cyclosporine A [19] on zebrafish heart. As the first organ in zebrafish to develop and function [20], the heart begins to develop as early as at 24 h post-fertilization (hpf) [7].…”

Cysteamine affects skeletal development and impairs motor behavior in zebrafish

“…The downregulation of cell cycle key genes ccne1, cdk2, cdk6, and the upregulation of ccnd1 suggested that cell proliferation was also seriously affected by the disordered expression of these genes. According to previous publications, excessive accumulation of ROS leads to oxidative stress, affects mitochondrial function, hinders cell proliferation, and further promotes cell apoptosis (Bai et al, 2021;Wan et al, 2021). Therefore, the apoptosis of zebrafish skeletal cells exposed to cysteamine was probably mediated by oxidative stress.…”

Identification of key pharmacodynamic markers of American ginseng against heart failure based on metabolomics and zebrafish model

“…The heart of zebrafish larvae was then imaged with inverted microscopy (Olympus IX83, Japan) (Supplementary Figure S1). The pericardial area, the venous congestion area (Lu et al, 2020), and the distance between the sinus venosus and bulbus arteriosus (SV-BA) of the heart (Wan et al, 2021) were counted by Image Pro Plus5.1 software. The data were analyzed by one-way analysis of variance (ANOVA) using GraphPad Prism 6.01 software and expressed as the mean ± standard error (SE).…”