Zebrafish larvae were used to further understand the liver toxicity of nux vomica. The histopathology, protein expression, and gene expression were assessed to confirm apoptosis in the liver, and then, profiles of the metabolites in zebrafish were investigated by untargeted metabolomic assessment to understand the potential toxicity mechanism of nux vomica. Histopathological observations showed that nux vomica caused damage to liver cells. Western blot results indicated increased expression of activated caspase3, and the result of real-time polymerase chain reaction showed a significant increase in the expression level of caspase-3, caspase-8, and caspase-9 genes (p < 0.05) compared with the control group. The liver injury from nux vomica was linked to the downregulation of amino acid (e.g., proline and alanine) and fatty acid (e.g., palmitoleic acid) metabolism and upregulation of some other fatty acid (e.g., arachidic acid) and purine (e.g., xanthine and uric acid) metabolism. The hepatotoxicity of nux vomica resulted from metabolic pathway disturbances, including small molecules involved in energy, purine, lipids, and amino acid metabolism.
Get full access to this article
View all access options for this article.
References
1.
YinW, WangT-S, YinF-Z, CaiB-C. Analgesic and anti-inflammatory properties of brucine and brucine N-oxide extracted from seeds of Strychnos nux-vomica. J Ethnopharmacol, 2003; 88:205–214.
2.
DengXK, YinW, LiWD, YinFZ, LuXY, ZhangXC, et al.The anti-tumor effects of alkaloids from the seeds of Strychnos nux-vomica on HepG2 cells and its possible mechanism. J Ethnopharmacol, 2006; 106:179–186.
3.
KhanM, GargA, SrivastavaSK, DarokarMP. A cytotoxic agent from Strychnos nux-vomica and biological evaluation of its modified analogues. Med Chem Res, 2011; 21:2975–2980.
4.
GoldbergAM.Animals and alternatives: societal expectations and scientific need. ATLA, 2004; 32:545–551.
5.
Caldwell SKCaJS. Fulfilling the promise: drug discovery in the post-genomic era. 2003; 8:168–174.
6.
DrewsJ.Drug Discovery: A Historical Perspective. Science, 2000; 287:1960–1964.
7.
PugsleyMK, GallacherDJ, TowartR, AuthierS, CurtisMJ. methods in safety pharmacology in focus. J Pharmacol Toxicol Methods, 2008; 58:69–71.
8.
ChuJ, SadlerKC. New school in liver development: lessons from zebrafish. Hepatology, 2009; 50:1656–1663.
9.
WhiteRM, SessaA, BurkeC, BowmanT, LeBlancJ, CeolC, et al.Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell, 2008; 2:183–189.
10.
ZonLI, PetersonRT. In vivo drug discovery in the zebrafish. Nat Rev Drug Discov, 2005; 4:35–44.
11.
LiuJ, StainierDY. Zebrafish in the study of early cardiac development. Circulation Res, 2012; 110:870–874.
12.
BakkersJ.Zebrafish as a model to study cardiac development and human cardiac disease. Cardiovasc Res, 2011; 91:279–288.
13.
SpitsbergenJ.Imaging neoplasia in zebrafish. Nat Methods, 2007; 4:548–549.
14.
BlackburnJS, LiuS, RaimondiAR, IgnatiusMS, SalthouseCD, LangenauDM. High-throughput imaging of adult fluorescent zebrafish with an LED fluorescence macroscope. Nat Protoc, 2011; 6:229–241.
15.
Gibert MCTaACWY.Zebrafish as a genetic model in pre-clinical drug testing and screening. Curr Med Chem, 2013; 20:2458–2466.
16.
PetersonRT, MacraeCA. Systematic approaches to toxicology in the zebrafish. Ann Rev Pharmacol Toxicol, 2012; 52:433–453.
17.
SipesNS, PadillaS, KnudsenTB. Zebrafish—as an integrative model for twenty-first century toxicity testing. Birth Defects Res Part C, 2011; 93:256–267.
18.
McGrathP, LiC-Q. Zebrafish: a predictive model for assessing drug-induced toxicity. Drug Discov Today, 2008; 13:394–401.
19.
RekhaRD, AmaliAA, HerGM, YehYH, GongHY, HuSY, et al.Thioacetamide accelerates steatohepatitis, cirrhosis and HCC by expressing HCV core protein in transgenic zebrafish Danio rerio. Toxicology, 2008; 243:11–22.
20.
FlemingA, JankowskiJ, GoldsmithP. In vivo analysis of gut function and disease changes in a zebrafish larvae model of inflammatory bowel disease: a feasibility study. Inflamm Bowel Dis, 2010; 16:1162–1172.
21.
CinarogluA, GaoC, ImrieD, SadlerKC. Activating transcription factor 6 plays protective and pathological roles in steatosis due to endoplasmic reticulum stress in zebrafish. Hepatology, 2011; 54:495–508.
22.
PasseriMJ, CinarogluA, GaoC, SadlerKC. Hepatic steatosis in response to acute alcohol exposure in zebrafish requires sterol regulatory element binding protein activation. Hepatology, 2009; 49:443–452.
23.
SongYN, ZhangH, GuanY, PengJH, LuYY, HuYY, et al.Classification of traditional Chinese medicine syndromes in patients with chronic hepatitis B by SELDI-Based proteinChip analysis. Evidencebased Complement Altern Med, 2012; 2012:626320.
24.
ZhangFT, ZhuZH, TongXR, et al.Mixed linear model approaches of association mapping for complex traits based on omics variants. Sci Rep, 2015; 5:10298.
25.
HarrillAH, RusynI. Systems biology and functional genomics approaches for the identification of cellular responses to drug toxicity. Expert Opin Drug Metab Toxicol, 2008; 4:1379–1389.
26.
JacobsA.An FDA perspective on the nonclinical use of the X-Omics technologies and the safety of new drugs. Toxicol Lett, 2009; 186:32–35.
27.
LiC, LiP, TanYM, LamSH, ChanEC, GongZ. Metabolomic characterizations of liver injury caused by acute arsenic toxicity in zebrafish. PLoS One, 2016; 11:e0151225.
28.
JeremyK. NicholsonJC, JohnC.Lindon,Elaine Holmes. Metabonomics a platform for studying drug toxicity and gene function. Nat Rev Drug Discov, 2002; 1:153–161.
29.
WilsonID, NicholsonJK, Castro-PerezJ, GrangerJH, JohnsonKA, SmithBW, et al.High resolution “Ultra Performance” liquid chromatography coupled to oa-TOF mass spectrometry as a tool for differential metabolic pathway profiling in functional genomic studies. J Proteome Res, 2005; 4:591–598.
30.
GowdaGA, ZhangS, GuH, AsiagoV, ShanaiahN, RafteryD. Metabolomics-based methods for early disease diagnostics. Expert Rev Mol Diagn, 2008; 8:617–633.
31.
WantEJ, MassonP, MichopoulosF, WilsonID, TheodoridisG, PlumbRS, et al.Global metabolic profiling of animal and human tissues via UPLC-MS. Nat Protoc, 2013; 8:17–32.
32.
ZhaoCJ, TianJ, WangJ, FengY, NiY, FanJ, et al.Zebrafish model for assessing induced organ toxicity by Strychnos nux-vomica. J Tradit ChinMed, 2016; 36:522–529.
33.
AmaliAA, RekhaRD, LinCJ, WangWL, GongHY, HerGM, et al.Thioacetamide induced liver damage in zebrafish embryo as a disease model for steatohepatitis. J Biomed Sci, 2006; 13:225–232.
34.
Saera-VilaA, KishPE, LouieKW, GrzegorskiSJ, KlionskyDJ, KahanaA. Autophagy regulates cytoplasmic remodeling during cell reprogramming in a zebrafish model of muscle regeneration. Autophagy, 2016; 12:1864–1875.
35.
NegronJF, LockshinRA. Activation of apoptosis and caspase-3 in zebrafish early gastrulae. Dev Dyn, 2004; 231:161–170.
36.
QiuYP, CaiGX, SuMM, ChenTL, ZhengXJ, XuY, et al.Serum metabolite profiling of human colorectal cancer using GC-TOFMS and UPLC-QTOFMS. J Proteome Res, 2008; 8:4844–4850.
37.
WangJH, ChenWL, LiJM, WuSF, ChenTL, ZhuYM, et al.Prognostic significance of 2-hydroxyglutarate levels in acute myeloid leukemia in China. Proc Natl Acad Sci U S A, 2013; 110:17017–17022.
38.
RandallT.Peterson, Brian A. Link, John E. Dowling aSLS. Small molecule developmental screens reveal the logic and timing of vertebrate development. PNAS, 2000; 97:12965–12969.
39.
RubinsteinAL.Zebrafish assays for drug toxicity screening. Expert Opin Drug Metab Toxicol, 2006; 2:231–240.
40.
RennekampAJ, PetersonRT. 15 years of zebrafish chemical screening. Curr Opin Chem Biol, 2015; 24:58–70.
41.
GoldstoneJV, McArthurAG, KubotaA, ZanetteJ, ParenteT, JonssonME, et al.Identification and developmental expression of the full complement of Cytochrome P450 genes in Zebrafish. BMC Genomics, 2010; 11:643.
42.
YeG, ChenY, WangHO, YeT, LinY, HuangQ, et al.Metabolomics approach reveals metabolic disorders and potential biomarkers associated with the developmental toxicity of tetrabromobisphenol A and tetrachlorobisphenol A. Sci Rep, 2016; 6:35257.
43.
JosephA, LeeT, MolandCL, BranhamWS, FuscoeJC, LeakeyJE, et al.Effect of (+)-usnic acid on mitochondrial functions as measured by mitochondria-specific oligonucleotide microarray in liver of B6C3F1 mice. Mitochondrion, 2009; 9:149–158.
44.
WangY, ZhuW, WangD, TengM, YanJ, MiaoJ, et al.(1)H NMR-based metabolomics analysis of adult zebrafish (Danio rerio) after exposure to diniconazole as well as its bioaccumulation behavior. Chemosphere, 2017; 168:1571–1577.
45.
VachierI, ChanezP, BonnansC, GodardP, BousquetJ, ChavisC. Endogenous anti-inflammatory mediators from arachidonate in human neutrophils. Biochem Biophys Res Commun, 2002; 290:219–224.
46.
DuZ, ZhangY, WangG, PengJ, WangZ, GaoS. TPhP exposure disturbs carbohydrate metabolism, lipid metabolism, and the DNA damage repair system in zebrafish liver. Sci Rep, 2016; 6:21827.
