Bioactive chemical constituents, in vitro anti-proliferative activity and in vivo toxicity of the extract of Curcuma singularis Gagnep rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma singularis Gagnep is a Vietnamese medicinal plant which has been commonly used as a medicinal remedy in traditional and folk medicines for improving health as well as for treating some diseases, like rheumatoid arthritis, kidney failure. However, pharmacological effects, including anti-cancer activity and the safety of this plant has not been fully investigated. AIM OF THE STUDY: This study aimed to investigate the in vitro anti-growth activity of an extract derived from Curcuma singularis rhizome extract (CSE) against cell lines as well as determine its phytochemical composition. The other goal of our study was to assess the safety of CSE in rats. MATERIALS AND METHODS: The main constituents in the extract were identified and quantitatively analyzed. The in vitro cytotoxicity of CSE was evaluated in several cancer and normal cell lines. The apoptotic activity of CSE and the expression of the apoptosis-related genes were investigated in AGS cells to clarify the underlying molecular mechanisms. The in vivo toxicity of CSE was assessed via acute and subacute oral studies on Sprague-Dawley rats, respectively according to the guidelines 425 and 407 of the Organization for Economic Cooperation and Development (OECD). The drug-related toxicity signs, mortality, body and organ weights were recoreded during the experimental period. In addition, the selected hematological and biochemical parameters, and histological alterations were determined at the end of the subacute toxicity test. RESULTS: Germacrone, ar-turmerone, and curcumol were three sesquiterpene components found in the extract. CSE showed cytotoxic effects in different cancer cells, but had minimal effects on normal cells. Apoptosis in AGS cells was caused by CSE in a concentration-dependent pattern through increase of Bax/Bcl-2 ratio, and release of cytochrome c, which leads to activation of caspase-3/-7, caspase-9, as well as cleavage of PARP. In the acute toxicity test, no signs of toxicity and no mortality were recorded in rats at both doses of 1000 and 5000 mg/kg. In the subacute toxicity study, CSE showed no drug-related adverse effects on water and food consumption, body and organ weights. CSE at a dose of 1000 mg/kg slightly increased WBC and platelet values in female rats, while it increased WBC values in male rats in all tested doses. The decrease of total cholesterol and triglyceride levels were found in female rats treated CSE at doses of 250 or 500 mg/kg. In addition, the increase of serum ALT and AST levels in rats treated at the dose of 1000 mg/kg were noted. No significant changes in histopathological structures of kidneys, spleen, heart and lungs, except liver tissue with minor modifications was found. CONCLUSIONS: Our findings indicated that CSE exhibited in vitro anti-proliferative effects on AGS cells by mainly activating the caspase-dependent mitochondrial apoptotic pathway. CSE also showed in vivo toxicity signals at the dose of 1000 mg/kg with proven minor hepatic injuries, which should be avoided the high dose for prolonged use. Curcuma singularis rhizomes may be used as a chemotherapeutic agent for the treatment of gastric cancer with in vitro anti-cancer investigation and in vivo biological safety evaluation.

Investigation of bioactive chemical constituents and anti-cancer activity of ethanol extract of Curcuma singularis Gagnep rhizomes.

Curcuma singularis Gagnep is a Vietnamese medicinal plant which has been commonly used in traditional and folk medicines for the treatment of different diseases. The goals of the present study are to investigate chemical composition and anti-proliferative activity of Curcuma singularis rhizome extract (CSE). The in vitro cytotoxicity of CSE was evaluated using WST-1 and LDH assays. The apoptosis induction was determined using nuclei DAPI staining and FACS assays. The main compounds of extract were identified and quantitatively analyzed using the validated HPLC method. The extract showed cytotoxic effects in various liver and breast cancer cells but had minimal effects on normal cells. It induced apoptosis on both Hep3B and SKBR3 cells in a dose-dependent manner. In addition, three sesquiterpene compounds, such as germacrone (3.25 ± 0.32 mg/g), ar-turmerone (1.12 ± 0.24 mg/g), and curcumol (0.31 ± 0.12 mg/g) were found as the main components of CSE. This is the first report on the in vitro cytotoxic effect of Curcuma singularis rhizomes against cancer cells.