Developmental toxicity of Clerodendrum cyrtophyllum turcz ethanol extract in zebrafish embryo.

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 edema, pericardial edema 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 (ß-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-ß-D-glucopyranoside were identified from EE. EE exposure during 6-96 h post-fertilization (hpf) at doses ranging from 80 to 200 µg/mL increased embryo mortality and reduced hatching rate. EE exposure at 20 and 40 µg/mL until 72-120 hpf induced a series of malformations, including yolk sac edema, pericardial edema, 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 µg/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.

'The child that tiire doesn't give you, God won't give you either.' The role of Rotheca myricoides in Somali fertility practices.

The paper introduces the Baanashada Dumarka, a Somali fertility therapy carried out by a spirit medium, known locally as 'Alaqad. Baanashada is aimed at women whose fertility issues are believed to be caused by spirits. The study also explores a component of the Baanashada, namely, the use of tiire (Rotheca myricoides), or the butterfly bush. Although Rotheca myricoides is known to possess a number of medicinal components as confirmed by studies of modern science, so far, there exist no studies on its potential (or lack of) fertility effects. Hence, the alleged fertility benefits of the butterfly bush need examining. In 2008 a British Somali woman died of herbs placed in her cervix by a traditional healer in Somaliland. This piece of information indicated not only the role of herbal medicine in fertility practices, but also the popularity of traditional reproductive medicine beyond border, class or educational background. Yet, current research into Somali women's health focuses mainly on Female Genital Mutilation (FGM), examined often without the context of wider cultural practices. This paper, however, suggests that rituals, beliefs and material culture play a paramount role in women's practices. For example, as explored elsewhere, the wagar, a wooden and sacred object made of the African olive, is critical for fertility practices. The current paper illuminates further the significance of reproduction practices in Somali society and the potential continuity of traditions associated with the perpetuation of kinship. It concludes that fertility rituals are part of a wider context of interaction with sacred landscapes, objects and archaeological sites, often associated with past legends in the Horn of Africa.