Involvement of Nrf2-HO-1/JNK-Erk Signaling Pathways in Aconitine-Induced Developmental Toxicity, Oxidative Stress, and ROS-Mitochondrial Apoptosis in Zebrafish Embryos

Abstract

Aconitine (AC), one of the bioactive diterpenoid alkaloids extracted from Aconitum plants, is widely used in traditional herbal medicine to treat various diseases. Emerging evidence indicates that AC has attracted great interest for its wide cardiotoxicity and neurotoxicity. However, the toxic effects of AC on embryonic development and its underlying mechanisms remain unclear. Here, a developmental toxicity assay of AC was performed on zebrafish embryos from 4 to 96 h post fertilization (hpf), and its underlying mechanisms were discussed. AC exposure impaired the cardiac, liver, and neurodevelopment. Especially, a high dose of AC (7.27 and 8.23 μM) exposure resulted in malformations at 72 and 96 hpf, including reduced body length, curved body shape, pericardial edema, yolk retention, swim bladder and brain developmental deficiency, and degeneration of dopaminergic neurons. High-concentration AC exposure caused a deficient cardiovascular system with cardiac dysfunctions, increased heart rates at 72 and 96 hpf, and reduced locomotor behavior at 120 hpf. AC treatment significantly increased the ROS level and triggered cell apoptosis in the heart and brain regions of embryos at 96 hpf in 7.27 and 8.23 μM AC treatment zebrafish. Oxidative stress was confirmed by reduced levels of T-SOD activity associated with accumulation of lipid peroxidation in larvae. The expression levels of oxidative stress-related genes (Nrf2, HO-1, Cat, and Sod-1) Erk1/2 and Bcl-2 were significantly downregulated at 96 hpf. The expression pattern of JNK and mitochondrial apoptosis-related genes (Bad, Bax, Cyto C, Casp-9, and Casp-3) was significantly upregulated. Taken together, all these parameters collectively provide the first evidence of AC-induced developmental toxicity in zebrafish embryo/larvae through ROS-medicated mitochondrial apoptosis involving Nrf2/HO-1 and JNK/Erk pathways.

An Updated Meta-Analysis Based on the Preclinical Evidence of Mechanism of Aconitine-Induced Cardiotoxicity

“…Moreover, 21 studies were deleted because the outcome measurement was not of interest. Ultimately, 32 eligible articles ( Liang et al, 1991 ; Meng, 2006 ; Fu, 2007 ; Liu, 2007 ; Wang, 2007 ; Wang et al, 2007 ; Zhang, 2007 ; Xu, 2008 ; Liu, 2009 ; Deng, 2010 ; Fang et al, 2012 ; Zhang et al, 2012 ; Wang, 2013 ; Zhou et al, 2013 ; Sun et al, 2014 ; Yu, 2015 ; Cui et al, 2018 ; Gao, 2018 ; Gao et al, 2018 ; Hu et al, 2018 ; Li et al, 2018 ; Zhang et al, 2018 ; Liu et al, 2019 ; Wang M. et al, 2020 ; Li et al, 2020 ; Peng et al, 2020 ; Zhang et al, 2020 ; Zhou et al, 2020 ; Xia et al, 2021 ; Yang et al, 2021 ; Ye et al, 2021 ; Wang S. et al, 2022 ) were identified for further evaluation, data extraction, and analysis ( Figure 2 ).…”

“…Of them, the animals used in three studies ( Liang et al, 1991 ; Zhang et al, 2012 ; Sun et al, 2014 ) comprised Sprague–Dawley (SD) and Wistar rats, and two studies ( Deng, 2010 ; Wang, 2013 ) guinea pigs. The weight of rats ranged from 150 to 350 g, and the weight of guinea pigs ranged from 300 to 350 g. Among which, zebrafish embryos were employed in seven studies ( Fang et al, 2012 ; Cui et al, 2018 ; Liu et al, 2019 ; Wang S. et al, 2020 ; Li et al, 2020 ; Xia et al, 2021 ; Ye et al, 2021 ), H9c2 cell in nine studies ( Gao, 2018 ; Gao et al, 2018 ; Hu et al, 2018 ; Zhang et al, 2018 ; Li et al, 2020 ; Peng et al, 2020 ; Zhang et al, 2020 ; Yang et al, 2021 ; Wang W. et al, 2022 ), primary cardiomyocytes of SD rats in 10 studies ( Meng, 2006 ; Fu, 2007 ; Wang, 2007 ; Wang et al, 2007 ; Zhang, 2007 ; Xu, 2008 ; Liu, 2009 ; Zhou et al, 2013 ; Yu, 2015 ; Li et al, 2018 ), primary cardiomyocytes of Wistar rats in three studies ( Liu, 2007 ; Sun et al, 2014 ; Zhou et al, 2020 ). Meanwhile, three studies chose primary cardiomyocytes of guinea pigs ( Wang, 2013 ) and AC-16 cells ( Cui et al, 2018 ; Zhou et al, 2020 ).…”

“…A large number of cardiomyocyte apoptosis is also considered a crucial marker for evaluating the cardiotoxicity of compounds ( Hu et al, 2018 ). In our study, the apoptosis rate was conducted in seven studies ( Gao, 2018 ; Hu et al, 2018 ; Zhang et al, 2018 ; Liu et al, 2019 ; Li et al, 2020 ; Xia et al, 2021 ; Yang et al, 2021 ). The meta-analysis of two studies ( Gao, 2018 ; Zhang et al, 2018 ) showed that compared with the control group, the apoptosis rate prominently surged and boomed in a concentration-dependent manner [ n = 18, SMD = 20.01, 95% CI (10.86 to 29.15), p < 0.0001; heterogeneity χ 2 = 1.68, df = 1 ( p = 0.20), I 2 = 40%] ( Figure 3D ).…”

Silibinin Alleviates Muscle Atrophy Caused by Oxidative Stress Induced by Cisplatin through ERK/FoxO and JNK/FoxO Pathways

“…Following Nrf2 nuclear translocation, it can positively regulate genes containing antioxidant response elements (ARE) in the promoter. The proteins encoded by these genes can participate in many protective reactions in the body, including antioxidant defense, protein degradation, cell growth, and DNA repair [ 24 ]. This may explain why the ROS content in the DDP-treated mice was significantly higher than that in other treatment groups in our study.…”

“…The MAPK family is often involved in cell apoptosis [ 22 , 23 ]. Furthermore, JNK/ERK related to oxidative stress has been reported to inhibit the nuclear factor erythrocyte 2-related factor 2 transcription factor (Nrf2) during apoptosis [ 24 ]. Nrf2 plays an important role in the activation of antioxidant enzymes and regulates the production of intracellular ROS.…”