Cytotoxicity and Toxicity Evaluation of Xanthone Crude Extract on Hypoxic Human Hepatocellular Carcinoma and Zebrafish (Danio rerio) Embryos

Abstract

Xanthone is an organic compound mostly found in mangosteen pericarp and widely known for its anti-proliferating effect on cancer cells. In this study, we evaluated the effects of xanthone crude extract (XCE) and α-mangostin (α-MG) on normoxic and hypoxic human hepatocellular carcinoma (HepG2) cells and their toxicity towards zebrafish embryos. XCE was isolated using a mixture of acetone and water (80:20) and verified via high performance liquid chromatography (HPLC). Both XCE and α-MG showed higher anti-proliferation effects on normoxic HepG2 cells compared to the control drug, 5-fluorouracil (IC50 = 50.23 ± 1.38, 8.39 ± 0.14, and 143.75 ± 15.31 μg/mL, respectively). In hypoxic conditions, HepG2 cells were two times less sensitive towards XCE compared to normoxic HepG2 cells (IC50 = 109.38 ± 1.80 μg/mL) and three times less sensitive when treated with >500 μg/mL 5-fluorouracil (5-FU). A similar trend was seen with the α-MG treatment on hypoxic HepG2 cells (IC50 = 10.11 ± 0.05 μg/mL) compared to normoxic HepG2 cells. However, at a concentration of 12.5 μg/mL, the α-MG treatment caused tail-bend deformities in surviving zebrafish embryos, while no malformation was observed when embryos were exposed to XCE and 5-FU treatments. Our study suggests that both XCE and α-MG are capable of inhibiting HepG2 cell proliferation during normoxic and hypoxic conditions, more effectively than 5-FU. However, XCE is the preferred option as no malformation was observed in surviving zebrafish embryos and it is more cost efficient than α-MG.

Inhibition of Seizure-Like Paroxysms and Toxicity Effects of Securidaca longepedunculata Extracts and Constituents in Zebrafish Danio rerio

“…A xanthone from G. mangostana, α-mangostin, has been found to be toxic and teratogenic in zebrafish embryos. − …”

Mangosteen Garcinia mangostana L. simplicia effect on bone structure and behaviour of Wader Fish Rasbora lateristriata (Bleeker, 1854) Embryo

“…This finding is in line with the research result conducted by Jose et al (2016), which reported the occurrence of Growth retardation, movement abnormalities, and, malformation of zebrafish embryos head after exposure to mangosteen leaves and stem-bark lyophilized water extract. In addition, research conducted by Fazry et al (2018) also showed the presence of a bent tail in the zebrafish embryos treated with α-mangostin. This indicates that some of the components contained in mangosteen pericarp or other parts of the plant could cause different effects on the embryonic development of different fish species.…”

Zebrafish as a Successful Animal Model for Screening Toxicity of Medicinal Plants

“…From the egg phase, zebrafish embryos can survive by ingesting yolk and can visibly be tested for malformation for a few days in a single well of a microplate [ 32 ]. When the embryo is not clumped, has a cardiac beat, has fully shaped body sections, and the tail is detached from the yolk, the standard endpoints of embryogenesis have been said to have occurred [ 28 ]. The main characteristics of embryogenesis and the genetic basis of growth in zebrafish have been widely researched [ 125 ].…”

“…One of the endpoints of embryonic growth in zebrafish is when the tail is detached entirely from the yolk [ 28 ], which can be affected by toxicants. The tail emerges during the larval stages from the primordium in the ventral fin fold, which coincides with a gap in the melanophore line immediately before the back tip of the notochord [ 149 ].…”

“…For example, various compound tests (e.g., measurement of drug cardiotoxicity) have been conducted to assess drug effects in zebrafish efficiently [ 11 ]. Additionally, for the last two decades, zebrafish have been used to study angiogenesis, metastasis, anticancer drug screening, and an assessment of drug toxicity [ 28 ]. In summary, the cell structural and biochemical similarities between humans and zebrafish enable rapid forecasting of the possible impacts of chemical and other substances on human communities.…”