Liver Fatty Acid Binding Protein Deficiency Provokes Oxidative Stress, Inflammation, and Apoptosis-Mediated Hepatotoxicity Induced by Pyrazinamide in Zebrafish Larvae

Abstract

g Pyrazinamide (PZA) is an essential antitubercular drug, but little is still known about its hepatotoxicity potential. This study examined the effects of PZA exposure on zebrafish (Danio rerio) larvae and the mechanisms underlying its hepatotoxicity. A transgenic line of zebrafish larvae that expressed enhanced green fluorescent protein (EGFP) in the liver was incubated with 1, 2.5, and 5 mM PZA from 72 h postfertilization (hpf). Different endpoints such as mortality, morphology changes in the size and shape of the liver, histological changes, transaminase analysis and apoptosis, markers of oxidative and genetic damage, as well as the expression of certain genes were selected to evaluate PZA-induced hepatotoxicity. Our results confirm the manner of PZA dose-dependent hepatotoxicity. PZA was found to induce marked injury in zebrafish larvae, such as liver atrophy, elevations of transaminase levels, oxidative stress, and hepatocyte apoptosis. To further understand the mechanism behind PZA-induced hepatotoxicity, changes in gene expression levels in zebrafish larvae exposed to PZA for 72 h postexposure (hpe) were determined. The results of this study demonstrated that PZA decreased the expression levels of liver fatty acid binding protein (L-FABP) and its target gene, peroxisome proliferator-activated receptor ␣ (PPAR-␣), and provoked more severe oxidative stress and hepatitis via the upregulation of inflammatory cytokines such as tumor necrosis factor alpha (TNF-␣) and transforming growth factor ␤ (TGF-␤). These findings suggest that L-FABP-mediated PPAR-␣ downregulation appears to be a hepatotoxic response resulting from zebrafish larva liver cell apoptosis, and L-FABP can be used as a biomarker for the early detection of PZA-induced liver damage in zebrafish larvae. P yrazinamide (PZA) is an important first-line drug in tuberculosis (TB) combination chemotherapy and is used during the initial 2 months of treatment for its remarkable sterilizing activity (1). Hepatotoxicity is the major adverse effect of PZA and usually occurs in the first 2 months of treatment (2). Previously reported studies showed a high incidence of hepatotoxicity with a high dosage of 40 to 70 mg/kg of body weight and a low rate of liver injury with a daily dose of Ͻ35 mg/kg (3). Although a reduction of the clinical dose considerably decreased the rate of PZA-induced hepatotoxic side effects, PZA is still considered to induce higher hepatotoxicity than other first-line antituberculosis drugs (4, 5). PZA hepatotoxicity cannot be ignored; however, little is known about the exact mechanism of PZA-induced hepatotoxicity.Previous research, by using microarray analyses and proteomics studies, found that metabolism, inflammation, and oxidative stress pathways were probably involved in PZA-induced hepatotoxicity (6, 7). A recent investigation showed that the metabolite 5-hydroxypyrazinoic acid (5-OH-PA) is responsible for PZAinduced hepatotoxicity (8). Despite an increasing number of researchers who are trying to reevaluate the hepatotoxic potentia...

Triptolide‐induced hepatotoxicity via apoptosis and autophagy in zebrafish

“…They were obtained from the Biology Institute of Shandong Academy of Sciences. They were raised and staged as previously described (Zhang et al, ).…”

“…The protocol to determine the liver area was as described previously (Zhang et al, ). After the larvae were anesthetized with 1 mg/mL tricaine, the larvae were fixed in 4% paraformaldehyde.…”

“…The selection of liver location was based on the fluorescence under a fluorescence microscope. The protocol for hematoxylin‐eosin staining was as described previously (Zhang et al, ). The fixed larvae were stained with hematoxylin and eosin.…”

Epidemiological, clinical and mechanistic perspectives of tuberculosis in older people

“…In DM with poor metabolic control, the hyperglycaemia and advanced glycation end‐products are strongly conducive to the development of oxidative stress and mitochondrial dysfunction, often culminating in disease complications resulting in organ or system dysfunction . Similarly, oxidative stress and mitochondrial dysfunction have also been shown to be responsible for toxicities incurred by isoniazid, pyrazinamide, aminoglycosides and fluoroquinolones in different experimental models . Due to the shared pathogenetic mechanisms in organ injury, pertaining to poorly managed DM and toxicities incurred by anti‐tuberculosis drugs, the aftermath could be seriously amplified in terms of morbidity and even mortality .…”

“…62 Similarly, oxidative stress and mitochondrial dysfunction have also been shown to be responsible for toxicities incurred by isoniazid, pyrazinamide, aminoglycosides and fluoroquinolones in different experimental models. [63][64][65][66] Due to the shared pathogenetic mechanisms in organ injury, pertaining to poorly managed DM and toxicities incurred by anti-tuberculosis drugs, the aftermath could be seriously amplified in terms of morbidity and even mortality. 67 In the geriatric population, as oxidative stress and mitochondrial dysfunction are also underscored mechanisms of the ageing process, with chronic inflammation as the pathogenetic basis in many 'degenerative' disorders, quite like DM, it would be reasonable to anticipate a similar interacting scenario resulting in antituberculosis drug toxicities among the geriatric patients.…”