Developmental Toxicity of Clerodendrum cyrtophyllum Turcz Ethanol Extract in Zebrafish Embryo

[en] ETHNOPHARMACOLOGICAL RELEVANCE: Clerodendrum cyrtophyllum Turcz has been used in traditional medicine for the treatment of various diseases. In spite of its therapeutic applications, research on its toxicity and teratogenicity is still limited. AIM OF THE STUDY: The study aimed to investigate the developmental toxicity of the ethanol extract of C. cyrtophyllum (EE) in zebrafish embryo model. MATERIAL AND METHODS: Major compounds from crude ethanol extract of Clerodendron cyrtophyllum Turcz leaves were determined using HPLC-DAD-Orbitrap-MS analysis. The developmental toxicity of EE were investigated using zebrafish embryo model. Zebrafish embryos at 6 h post-fertilization (hpf) were treated with EE at different concentrations. Egg coagulation, mortality, hatching, yolk sac oedema, pericardial oedema and teratogenicity were recorded each day for during a 5-day exposure. At time point 120 hpf, body length, pericardial area, heartbeat and yolk sac area were assessed. In order to elucidate molecular mechanisms for the developmental toxicity of EE, we further evaluated the effects of the EE on the expression of genes involved on signaling pathways affecting fish embryo's development such as heart development (gata5, myl7, myh6, has2, hand2, nkx 2.5), oxidative stress (cat, sod1, gpx4, gstp2), wnt pathway (beta-catenin, wnt3a, wnt5, wnt8a, wnt11), or cell apoptosis (p53, bax, bcl2, casp3, casp8, casp9, apaf-1, gadd45bb) using qRT-PCR analysis. RESULTS: Our results demonstrated that three major components including acteoside, cirsilineol and cirsilineol-4'-O-beta-D-glucopyranoside were identified from EE. EE exposure during 6- 96 h post-fertilization (hpf) at doses ranging from 80 - 200 mug/ml increased embryo mortality and reduced hatching rate. EE exposure at 20 and 40 mug/ml until 72 - 120 hpf induced a series of malformations, including yolk sac oedema, pericardial oedema, spine deformation, shorter body length. Based on two prediction models using a teratogenic index (TI), a 25% lethality concentration (LD25) and the no observed-adverse-effect level (NOAEL), EE is considered as teratogenic for zebrafish embryos with TI (LC50/EC50) and LD25/NOAEC values at 96 hpf reaching 3.87 and 15.73 respectively. The mRNA expression levels of p53, casp8, bax/bcl2, gstp2, nkx2.5, wnt3a, wnt11, gadd45bb and gata5 were significantly upregulated by EE exposure at 20 and 40 mug/ml while the expression of wnt5, hand2 and bcl2 were downregulated. CONCLUSIONS: These results provide evidence for toxicity effects of EE to embryo stages and provide an insight into the potential toxicity mechanisms on embryonic development.

Research Center/Unit :

GIGA-I3 - Giga-Infection, Immunity and Inflammation - ULiège
CART - Centre Interfacultaire d'Analyse des Résidus en Traces - ULiège

Disciplines :

Pharmacy, pharmacology & toxicology

Author, co-author :

Nguyen, Thu Hang

Nguyen, Phuc-Dam

Quetin-Leclercq, Joëlle

Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > I3-Laboratory for Organogenesis and Regeneration

Ly Huong, Thu Hang

Pham, The Hai

Kestemont, Patrick

Language :

English

Title :

Developmental Toxicity of Clerodendrum cyrtophyllum Turcz Ethanol Extract in Zebrafish Embryo

Publication date :

2021

Journal title :

Journal of Ethnopharmacology

ISSN :

0378-8741

eISSN :

1872-7573

Publisher :

Elsevier, Switzerland

Volume :

267

Pages :

113538

Peer reviewed :

Peer Reviewed verified by ORBi

Additional URL :

https://authors.elsevier.com/c/1c3nw1M9M1Mskx

Funders :

ARES - Académie de Recherche et d'Enseignement Supérieur [BE]

Commentary :

2020/11/05

Available on ORBi :

since 13 November 2020

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