Background: Alcoholic liver disease (ALD) is a major health problem referring to the collection of liver damage caused by excessive alcohol intake. The search for effective and safe alternatives for compounds from plants to protect the liver from extensive damages and delay the progress to a disease is still a big effort done in the scientific community. 2,3,5,6-Tetramethylpyrazine (TMP) is a compound found in a Chinese herbal medicinal plant, Ligusticum chuanxiong Hort and in some other plants. Objective: This study was done to assess the hepatoprotective effects of TMP against ALD using histopathological analysis of zebrafish livers subjected to different exposure groups. TMP has been mainly used for the treatment of cardio- and cerebrovascular diseases due to its antioxidant, anti-inflammatory, and antiapoptotic properties. Methodology: Adult male zebrafish were exposed to three TMP concentrations (40, 60, and 80 mg/L TMP) and to 1% v/v of ethanol. The dissected livers of the zebrafish were processed for fixing on glass slides using the H&E stains and were observed under the light compound microscopes for scoring. The safety of the TMP to the early life stages of the zebrafish was tested using the Zebrafish Embryotoxicity Test (ZFET). Results: Results showed that TMP was able to dose-dependently decrease mean scores for the four parameters diagnostic of ALD, i.e., steatosis, inflammation, cell death, and ballooning degeneration. These scores were comparable to those of the untreated group (no ethanol + no treatment) and positive control (ethanol + Hepasil DTXTM), with all groups' scores being statistically different from those of the negative control group (ethanol + no treatment) (p<0.05). Results for the ZFET showed that incidence of embryo mortality as well as teratogenic malformations of embryos exposed to TMP were significantly lower compared to the positive control group. Conclusion The hepatoprotective role of TMP was implied because anomalies such as cholestasis, vessel congestion, and hemorrhage were only observed in the ethanol-treated group and not in the other groups. In the analysis of the early development of the embryos using the Zebrafish Embryotoxicity Test (ZFET), TMP was found to be non-toxic and non-teratogenic at concentrations used for liver treatment. These initial findings on TMP provided justification for its plausibility as a hepatoprotective compound against alcoholic liver diseases (ALD).
Liver Diseases, Alcoholic

Background: The escape of polyethylene microbeads from waste-water treatment facilities to aquatic habitats has been a major concern by scientific communities due to the adverse effects on aquatic organisms as well as the well-being of the marine and terrestrial ecosystems. Objective: This study was conducted to evaluate the embryotoxic and teratogenic effects of polyethylene microbeads on the early development of the zebrafish Danio rerio using the Fish Embryo Acute Toxicity Test (FET). Methodology: Sixty (60) zebrafish embryos were exposed to polyethylene microbead suspensions (PE-MBS) of 20 μg/L, 200 μg/L, and 2000 μg/L concentrations. Using FET, the toxicological endpoints (i.e., egg coagulation, lack of somite formation, non-detachment of tail, and lack of heartbeat) were observed every 24 hours until the 96th-hour exposure. Hatching of the embryo from the chorion was observed from 48-96 hpf (hours-post fertilization), and at least four parameters of teratogenicity (i.e., edema of the pericardium and yolk sac, bent axis, tail curvature, and collapsed swim bladder) was observed at 144 hpf. Results: Significant differences between means and variances in the embryotoxic and teratogenic effects were observed for all treatment groups in relation to the negative control (reconstituted water). The emulsifier control (0.01% Tween 80, p-value=0.9), the solvent control (1% DMSO, p-value = 0.9), and the 20 μg/L PE-MBS (p-value = 0.92) did not significantly differ with the negative control group. However, the positive control (5% ethanol, pvalue= 7.8) and 200 μg/L (p-value = 1.1), and 2000 μg/L (p-value = 1.48) of PE-MBS were significantly embryotoxic and teratogenic to the developing organism. Conclusion The high concentrations of PE-MBS (200 μg/L and 2000 μg/L) may induce early hatching, mortality, and malformations. Tukey Kramer post hoc test substantiated that PE-MBS toxicity is dose-dependent since embryotoxicity and teratogenicity increase at higher concentrations. Further studies should be conducted to know more about the adverse effects of polyethylene microbeads on the development, physiology, and genomics of freshwater fishes.
Zebrafish