[논문]Preventive effects of sea cucumber (Apostichopus japonicus) ethanol extract on palmitate-induced vascular injury in vivo

Zhang, Chunying; Cha, Seon-Heui

doi:10.47853/fas.2022.e9

[국내논문] Preventive effects of sea cucumber (Apostichopus japonicus) ethanol extract on palmitate-induced vascular injury in vivo 원문보기

Fisheries and aquatic sciences, v.25 no.2, 2022년, pp.90 - 100

Zhang, Chunying (Department of Marine Bio and Medical Sciences, Hanseo University) , Cha, Seon-Heui (Department of Marine Bio and Medical Sciences, Hanseo University)

Abstract ▼ AI-Helper

Cardiovascular diseases (CVDs) have posed serious public health problems, accounting for nearly 30% of mortality worldwide and their incidence is still increasing. Therefore, new treatment resources are necessary to prevent or manage the ever-increasing population of patients with CVDs. Sea cucumber is well known for its medical and health benefit effects, but it is not well known what/how effect it has on vascular disease. In the present study, we examined the protect effect of sea cucumber, Apostichopus japonicus 80% ethanol extract (AJE) on zebrafish embryo with the stimulation of free fatty acid, palmitate (PA). In vivo study showed that AJE can attenuate PA-induced toxicity through relieving the rapid heartbeat, increasing the survival rate and reducing the malformation in both wild type and Tg (fli1a:eGFP) transgenic zebrafish lines. Additionally, compare with PA treated embryos, the yolk sac area, body length, axial vascular segment (AVS) and intersegmental vessel (ISV) of the co-treatment group of AJE and PA were comparable to the control group. Moreover, AJE lowered the expression of inducible nitric oxide synthase (iNOS), nitric oxide (NO) and inflammation-related genes induced by PA, and inhibited PA-induced vascular development disorders. Our data preliminarily verify that AJE could be a candidate resource for the prevention or therapy of CVDs.

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표/그림 (6)

표 Table 1. Primer sequences
표 Table 2. Phytotoxicity evaluation of different concentrations of AJE on zebrafish embryos
표 Table 3. Toxicity evaluation of different concentrations of PA on zebrafish embryos
그림 Fig. 1. Apostichopus japonicus extract (AJE) attenuates palmitate (PA)-induced toxicity in zebrafish embryos. Embryos were treated with 2 mM PA, or co-treated with 25 μg/mL AJE in egg water with 75 μM phenylthiourea (PTU) at 24–25 hpf. (a) Heart rates of zebrafish were measured at 75 hpf, the results are expressed as the beats/min. (b) survival rate of zebrafish were measured at 55 hpf and 75 hpf. (c) malformation were measured at 50 hpf and 70 hpf. (d) morphology was photographed at 55 hpf. (e) yolk sac area (mm2) and (f) body length (mm) of zebrafish were measured at 55 hpf. n = 10–50. Scale bar: 1 mm. *p < 0.05 and **p < 0.01. hpf, hours post fertilization.
그림 Fig. 2. Inhibitory effect of Apostichopus japonicus extract (AJE) on palmitate (PA)-induced nitric oxide (NO) and cell death in zebrafish larvae. 86 hpf larvae were incubated with diaminofluorophore 4-amino-5-methylamino-2’,7’-difluorofluorescein diacetate (DAF-FM DA, 5 mM) for 30 min and acridine orange (AO, 5 μg/mL) for 15 min to detect NO production (a, c) and cell death (b, d), respectively. The experiments were performed in triplicate. 86 hpf embryos were stained with 5 μM DAF-FM-DA (a) or 5 μg/mL AO (e) for 30 min at 28.5℃ in the dark. Scale bar: 1 mm. *p < 0.05, **p < 0.01 and ***p < 0.001. hpf, hours post fertilization.
그림 Fig. 3. Apostichopus japonicus extract (AJE) attenuated palmitate (PA)-induced toxicity on vascular morphogenesis in Tg (fli1a:eGFP) transgenic zebrafish. (a) Fluorescence images of Tg (fli1a:eGFP) transgenic zebrafish embryos at 60 hpf treated with phenylthiourea (PTU, control), PA and pretreatment of AJE with PA. The red horizontal and vertical rows in blue and yellow dotted boxes represent the partial view of axial vascular segment (AVS) and intersegmental vessel (ISV) length, respectively. (b) The length of AVS and ISV were measured by cellSens software. The relative mRNA expression of (c) Fli1, and (d) Lmo2 were analyzed by qPCR. All of measurements were performed in triplicate. Scale bar: 1 mm. *p < 0.05, **p < 0.01 and ***p < 0.001. hpf, hours post fertilization.

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