Vietnamese Medicinal Plant Research Articles

Identification of potential inhibitors of human aldose reductase from natural compounds of Vietnamese medicinal plants using in silico approaches

Identification of potential inhibitors of human aldose reductase from natural compounds of Vietnamese medicinal plants using in silico approaches

Screening Bioactive Compounds from Vietnamese Medicinal Plants as Inhibitors of CCR5 and CXCR4 Co-receptors to Prevent and Treat HIV-1 Infection by Molecular Docking Methods

Screening Bioactive Compounds from Vietnamese Medicinal Plants as Inhibitors of CCR5 and CXCR4 Co-receptors to Prevent and Treat HIV-1 Infection by Molecular Docking Methods

Curcuma aromatica rhizomes ethanol extract: A potential therapeutic agent against HepG2 Hepatoma cells

Curcuma aromatica, a Vietnamese medicinal plant, exhibits diverse therapeutic applications including blood loss, blood stasis, bloody urination, bloody vomiting, irregular menstruation, epilepsy, and cancer treatment. C. aromatica rhizomes ethanol extract and curcumol content have been shown strong inhibition of cancer growth. HepG2 cells were exposed to different doses of ethanol extract and curcumol extract. The degree of cell growth inhibition was determined using the WST-1 assay, and Hoechst 33258 spectrofluorometric assay, while the rate of cell death by apoptosis was assessed using the annexin-propidium iodide double-staining assay. Flow cytometry was employed to study the cell cycle, while the transwell assay was used to assess cell migration and invasion. The expression of bax and bcl-2 genes was analyzed using real-time reverse transcription-PCR. C. aromatica rhizomes ethanol extract (CE) consisted of a total phenolic content value of 15.02±0.18 (mg/g extract) and a curcumol level of 849.8 ± 23.73 (?g/g extract). The growth of HepG2 cells was suppressed in a dose-dependent manner by C. aromatica rhizomes ethanol extract. Following a 72-hour exposure to an (CE) concentration of 100 ?g/mL, the inhibition rate was measured to be 34.559 ± 0.456 %, the IC50 value was determined to be 44.79 ?g/mL and the inhibition of curcumol was 50.961± 0.641 %, and the IC50=93.48. The application of CE induced late apoptosis in HepG2 cells. The CE also induced programmed cell death (apoptosis) in HepG2 cancer cells, effectively suppressing their growth, movement, and invasion by controlling the synthesis of Bcl-2 mRNA. This work gives novel insights into the antihepatoma action of (CE) and establishes a foundation for the development of preliminary anticancer medications.

Carabrachiatanins A and B: Two New Phenylpropanoid-Substituted Catechins of Carallia brachiata Merr

Objectives: Carallia brachiata Merr. is a Vietnamese medicinal plant, used to treat mouth ulcers, tongue sores, sore throats, and malaria. This paper focuses on the chemical constituents of C. brachiata and their inhibition of NO production. Methods: The dried and minced sample was extracted with methanol. The crude extract was isolated by various chromatographic methods. The structures of the isolates were determined by HR-ESI-MS, NMR, and CD spectra. The NO inhibition activity was evaluated in LPS-activated RAW264.7 cells. Results: Two new catechin derivatives (carabrachiatanins A (1) and B (2)) and seven known proanthocyanidins (3-9) were isolated from C. brachiata by bioguided fractionation. All isolates significantly showed NO inhibition effects with IC50 values ranging from 13.72 ∼ 34.28 µM. Conclusions: Two new and seven known compounds were isolated from C. brachiata. This is the first report of compounds 3-9 from C. brachiata. Compounds 7 and 8 inhibited NO production with their IC50 values of 16.31 ± 0.61 µM and 16.53 ± 0.83 µM, respectively, compared to that of dexamethasone, which showed IC50 value of 14.94 ± 1.11 µM.

Knowledge, attitude and practice on the use of vietnamese medicinal plants for common diseases treatment among people in Phu Vang district, Thua Thien Hue province

Background: Vietnamese herbs are considered as a botanical resource, a precious medicinal source of our country, and are widely used in the residential community in general. Phu Vang district is a coastal plain and lagoon in Thua Thien Hue province, where people’s living standards are not high, health facilities lack equipment. Therefore, the needs for primary health care are necessary to be strengthened, especially in the treatment of common diseases using Vietnamese herbs to reduce costs and improve health for the people. However, the knowledge of the local residents about Vietnamese herbs preventing and treating common diseases at home has not been surveyed. Objectives: (1) To describe the knowledge, attitude and practice on the use of Vietnamese herbs used in the treatment of common diseases of residents in some communes of Phu Vang district. (2) To determine some factors related to the use of Vietnamese herbs in the treatment of common diseases among the residents. Methods: A cross-sectional study was conducted on 400 residents representing the heads of households in three communes of Phu Vang district, Thua Thien Hue province, Thua Thien Hue province. Descriptive statistical analysis and multivariable logistic regression were performed with SPSS software. Results: Knowledge: Good: 45.2%; Fair: 30.8%; Average: 9.0%; Poor: 2.0%. Attitude: 97.0% of residents preferred using Vietnamese herbs in the treatment of common diseases, and 96.6% of residents agreed to continue using Vietnamese herbs to treat similar diseases later. Practice: Good: 7.8%; Fair: 16.7%; Average: 25.3%; Poor: 50.2%. Receiving information about Vietnamese herbs and growing them at home gardens were two factors related to the use of Vietnamese herbs to treat common diseases (p < 0.05). Conclusion: The practice of using Vietnamese herbs to treat common diseases of the residents in Phu Vang district is still low, it is necessary to strengthen educational propaganda to raise the awareness and practice among residents on the prevention and treatment of diseases with Vietnamese herbs at home. Key words: knowledge, attitude, practice, Vietnamese herbs, common diseases, Phu Vang district.

Studies on hepatoprotective effects of Vietnamese medicinal plants

The liver is a major organ of the human and animal body, playing an important role in metabolism, detoxification and immunomodulation. When liver tissue is damaged, liver function will decrease, leading to liver diseases such as liver fibrosis, cirrhosis, and liver cancer. Medicinal plants and natural products have provided a source of new drug candidates for the prevention and treatment of liver diseases. This review is based on scientific publications on Vietnamese plants investigated for their hepatoprotective activity during the period from 1998 to 2022. 51 medicinal plants, 38 phytoconstituents, and 8 hepatoprotective herbal formulations of Vietnamese-origin have been reported to protect the liver from the harmful effects induced by hepatotoxins, typically CCl4 or paracetamol. Each plant's information, including its botanical name, family, part of the plant used, local name, chemical constituents, extracts or fractions used, dosage, model used, parameters obtained, histopathology, and results of hepatoprotective studies, is displayed. Also discussed is the possible mechanism of action of these hepatoprotective plants.

Phenolic Constituents of Elsholtzia blanda Benth

Objectives: Elsholtzia blanda Benth. is a Vietnamese medicinal plant, used to treat inflammation, cough, and flu. Therefore, this current study focused on the isolation of the active ingredients from this plant and their NO production inhibitory activity. Methods: E. blanda aerial parts were dried, minced, and ultrasonically extracted with methanol and further separated by various chromatographic methods. The chemical structures of isolates were revealed by nuclear magnetic resonance, high resolution lectrospray ionization mass spectrometry, and circular dichroism spectra. The nitric oxide inhibition effects were evaluated in lipopolysaccharide-activated RAW264.7 cells. Results: Ten phenolic components [1 new (1) and 9 known (2-10)] were isolated and their chemical structures determined. Compound 5 (innovanoside, IC50 = 19.23 ± 1.43 µM) and 6 (maltol- O- β-D-glucopyranoside, IC50 = 21.73 ± 1.27 µM) showed the highest activity, and 2-4 showed moderate inhibition (IC50 = 47.36 ∼ 57.29 µM). The other compounds were inactive with IC50 values >100 µM. Conclusions: One previously undescribed and 9 known phenolic components isolated from E. blanda have been screened for their nitric oxide inhibitory activity. Compounds 2-6 showed moderate to significant inhibition with IC50 values of 19.23∼57.29 µM.

Panaxindole, a novel indole alkaloid N-glucoside from the leaves of Panax vietnamensis Ha et Grushv. (Vietnamese ginseng).

Vietnamese ginseng (Panax vietnamensis Ha and Grushv., Araliaceae) is indigenous in the central highlands of Vietnam and the southernmost distribution in the Panax genus. Like other ginseng, Vietnamese ginseng is well known has been used as a tonic and for management of certain diseases in the traditional medicine. Nevertheless, it is noteworthy that in respect to the long history in use and systematic studied on Korean ginseng (P. ginseng), American ginseng (P. quinquefolius), Japanese ginseng (P. japonicus), and Chinese ginseng (P. notoginseng), the up-to-date published database on Vietnamese ginseng is relatively much less extensive. In our ongoing research on the promising Vietnamese medicinal plants, the present phytochemical investigation of the ethanol extract of the leaves of Panax vietnamensis led to the isolation of three compounds (1-3), including a new indole alkaloid N-glycoside (1) and two known compounds. Their structures were elucidated based on extensive physiochemical and chemical methods, especially the interpretation of NMR and MS spectra. The absolute configuration of 1 was determined based on the comparison of its experimental and theoretical ECD spectra along with NMR calculation. Compound 1 is naturally isolated N-glycoside, which is rarely found in natural products. The isolated compounds showed weak or no inhibitory activity against acetylcholinesterase enzyme (AChE).

Screening for in vitro inhibiting Nrf2 of some Vietnamese medicinal plants

Nuclear factor erythroid 2–related factor 2 (Nrf2) is vital in regulating cellular defenses against oxidative or environmental aggressors. Some studies reveal that increasing Nrf2 activity affects chemotherapy drug resistance in cancer; thus, Nrf2 has become an attractive target in cancer therapeutics research. Our study screened the effects of 52 methanol extracts from different parts of 24 medicinal plants used in traditional and folk medicine to evaluate the ability to inhibit Nrf2 and the cell viability by an in vitro model of Huh7 liver cancer cells at the crude extract concentration of 100 µg/mL. The study has successfully suggested a screening model for Nrf2 expression based on luciferase fluorescence assay of Huh7 cells transfected with the Nrf2 gene. The results of our study show that the Piper sarmentosum roots can inhibit nearly 90% of Nrf2 activity in cancer cells. We also found that the extracts of Vernonia amygdalina leaves, Phyllanthus amarus leaves, Zingiber zerumbet leaves, Hyptis suaveolens leaves and roots, Curcuma zedoaria leaves and roots, Luvunga scandens leaves, Helicteres hirsuta leaves, Oroxylum indicum stem and leaves, Lasia spinosa fruits and Crotalaria pallida leaves inhibited more than 60% of Nrf2 expression levels. Furthermore, the extracts from the leaves of Luvunga scandens and Curcuma zedoaria can cause toxicity on cancer cell of more than 70% at 100 µg/mL. From the study, we have the basis for further studies on natural active substances that have the effect of drug resistance by inhibiting the expression of Nrf2 on cancer cells.

Determination of Antioxidant, Cytotoxicity, and Acetylcholinesterase Inhibitory Activities of Alkaloids Isolated from Sophora flavescens Ait. Grown in Dak Nong, Vietnam.

Traditional/herbal medicine has gained increasing interests recently, especially in Asian countries such as Vietnam, due to its diverse therapeutic actions. In the treasure of Vietnamese medicinal plants, one of the potential herbs is the roots of Sophora flavescens Ait. (SF, "Kho sam" in Vietnamese). However, limited information has been reported on the Vietnamese SF compositions and their respective alkaloids' anti-acetylcholinesterase action. Thus, this study investigated the extractions, isolations, identifications, and in-vitro antioxidant, cytotoxicity, and acetylcholinesterase inhibitory activities, of the SF root extracts and their purified alkaloid compounds. To this end, four pure compounds were successfully isolated, purity-tested by HPLC, and structurally identified by spectroscopic techniques of FTIR, MS, and NMR. These compounds, confirmed to be oxysophocarpine, oxymatrine, matrine, and sophoridine, were then determined their therapeutic actions. The SF extracts and the compounds did not possess significant antioxidant activity using the DPPH and MDA assays, and cytotoxicity action using the MTT assay on the MDA-MB-231 breast cancer cell line. On the other hand, the SF total extract yielded a moderate acetylcholinesterase inhibition effect, with an IC50 of 0.1077 ± 0.0023 mg/mL. In summary, the SF extract demonstrated potential effects as an anti-acetylcholinesterase agent and could be further researched to become a pharmaceutical product for diseases related to acetylcholine deficiency, such as dementia.

Diversity of medicinal plant resources in Lai Son island, Kien Giang province

This study was conducted with the aim of assessing the diversity of medicinal plants in Lai Son Island, Kien Giang province, which is a scientific basis for more effectively using, managing, and preserving this medicinal plant resource in this province. In this study, the PRA (Participatory Rural Appraisal), field investigation, morphological comparison, and classification, with the aid of specialized medicinal plant books were used. The results showed a total of 353 species of medicinal plants belonging to 264 genera of 112 families in 4 divisions. Among them, Magnoliophyta was the most diverse division with 94.90% % species, 95.08% genera, and 90.18% families. Seven species were listed in “Red List of Vietnamese medicinal plants” (2006), “Vietnam Red Book” (2007), and the Decree 06/2019/ND-CP. The medicinal plant species were divided into ten life forms and distributed in six biotopes. The most species diversity was recorded in the natural forest on rocky mountain biotope. Ten parts of plants were used to medicate 36 disease types. Twenty-four species were commonly used by local people.

Penicillium digitatum as a Model Fungus for Detecting Antifungal Activity of Botanicals: An Evaluation on Vietnamese Medicinal Plant Extracts.

Medicinal plants play important roles in traditional medicine, and numerous compounds among them have been recognized for their antimicrobial activity. However, little is known about the potential of Vietnamese medicinal plants for antifungal activity. In this study, we examined the antagonistic activity of twelve medicinal plant species collected in Northern Vietnam against Penicillium digitatum, Aspergillus flavus, Aspergillus fumigatus, and Candida albicans. The results showed that the antifungal activities of the crude extracts from Mahonia bealei, Ficus semicordata, and Gnetum montanum were clearly detected with the citrus postharvest pathogen P. digitatum. These extracts could fully inhibit the growth of P. digitatum on the agar medium, and on the infected citrus fruits at concentrations of 300–1000 µg/mL. Meanwhile, the other tested fungi were less sensitive to the antagonistic activity of the plant extracts. In particular, we found that the ethanolic extract of M. bealei displayed a broad-spectrum antifungal activity against all four pathogenic fungi. Analysis of this crude extract by enrichment coupled with high-performance liquid chromatography revealed that berberine and palmatine are major metabolites. Additional inspections indicated berberine as the key compound responsible for the antifungal activity of the M. bealei ethanolic extract. Our study provides a better understanding of the potential of Vietnamese medicinal plant resources for combating fungal pathogens. This work also highlights that the citrus pathogen P. digitatum can be employed as a model fungus for screening the antifungal activity of botanicals.

Steroid glycosides isolated from Paris polyphylla var. chinensis aerial parts and paris saponin II induces G1/S-phase MCF-7 cell cycle arrest

In our previous research on Vietnamese medicinal plants, we found that the ethanolic extract of the aerial parts of Paris polyphylla var. chinensis exhibited cytotoxic effects in vitro in the MCF-7 human cancer cell line. Here, we used combined chromatographic separations to isolate six compounds including a new steroid glycoside, paripoloside A (3), and five known compounds, from the butanol extract of the aerial parts of P. polyphylla. We unambiguously elucidated their structures based on spectroscopic data (proton and carbon-13 nuclear magnetic resonance, heteronuclear single quantum coherence, heteronuclear multiple bond correlation, correlation spectroscopy, and high-resolution electrospray ionization mass spectroscopy data), and chemical reactions. Among the isolated compounds, paris saponin II (PSII) had the strongest cytotoxic effects against MCF-7 breast cancer cells. Interestingly, PSII significantly increased the expression of p53, p21, p27, and Bax protein levels and significantly suppressed the expression of cyclin D1 and retinoblastoma protein. These data suggest that PSII may induce G1/S phase cell cycle arrest and apoptosis pathway development in MCF-7 cells. Furthermore, the MCF-7 breast cancer cells mechanism of PSII was also investigated using molecular docking. Together, our results demonstrate that isolated compounds from P. polyphylla are promising candidates as breast cancer inhibitors.

Rotenoids and coumaronochromonoids from Boerhavia erecta and their biological activities: In vitro and in silico studies

Boerhavia erecta is a tropical plant that is widely used in folk medicine. Roots of this plant are applied as a diuretic, stomachic, anthelminthic, febrifuge, and expectorant. The whole plant has been used for treating gastrointestinal, liver, and infertility problems. More than 40 metabolites have been identified in its leaves, roots, and the aerial parts. Among them, isoflavonoids such as rotenoids and coumaronochromonoids are major components. Some of these compounds exhibited potent cytotoxicity, COX inhibition and opioid and cannabinoid receptor. However, little is known about the inhibition of α-glucosidase enzyme of these compounds. As part of our ongoing search for α-glucosidase inhibitors from Vietnamese medicinal plants, we conducted bioactive-guided isolation of the aerial part of Boerhavia erecta. Twelve known compounds were isolated and elucidated, including boeravinone M (1), boeravinone G (2), boeravinone P (3), boeravinone B (4), boeravinone E (5), boeravinone A (6), boeravinone H (7), 9-O-methyl-4-hydroxyboeravinone B (8), boeravinone T (9), boeravinone J (10), boeravinone C (11), and 10-demethylboeravinone C (12). Isolates 2–5, 7, and 12 showed potent α-glucosidase inhibition, which is stronger than acarbose, a positive control. Among them, compound 5 showed strongest inhibition with IC50 value of 85.1 µM. A kinetic study indicated that 5 was a mixed-type inhibitor. Compound 5 was determined to be a potential inhibitor of the α-glucosidase enzyme based on in silico molecular docking mode. Compounds were tested for cytotoxicity against K562 and antimicrobial activity against antibiotic-resistant, pathogenic bacteria Enterococcus faecium, Staphylococcus aureus, and Acinetobacter baumannii. Only compounds 4 and 12 showed moderate activity against K562 cell line. As regards antimicrobial activity, compounds 6 and 10 showed strong inhibition toward Enterococcus faecium. In silico studies of cytotoxicity and antimicrobial activity were also performed, indicating that in vitro and in silico data were consistent.

20(22)Z and 20(22)E Dammarane Saponins From the Roots of Panax pseudoginseng Wall.

Panax pseudoginseng Wall. is famous as a traditional Vietnamese medicinal plant used to promote health and aid in cancer treatment. From the roots of this plant, 1 new (1) and 4 known (2-5) Δ20(22)-dammarane-type triterpenoid glycosides were isolated by various chromatographic methods. Their chemical structures were determined as 3β,6α,12β-trihydroxydammarane-( Z)-20(22),24-diene 6 -O-β-D-glucopyranoside (1), 3β,6α,12β-trihydroxy-dammar-( E)-20(22),25-diene 6 -O-β-D-glucopyranoside (2, ginsenoside Rh4), ginsenoside Rg5 (3), 3β,12β-dihydroxydammarane-( E)-20(22),24-diene 6 -O-β-D-xylopyranosyl-(1→2)-β-D-glucopyranoside (4), and 3β,12β-dihydroxydammarane-( E)-20(22),24-diene 6 -O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (5). The E/ Z-Δ20(22) configurations in the dammarane compounds were further evidenced by 1D and 2D NMR data.

In vitro and in silico Screening of Bioactive Compounds from Jasminum subtriplinerve Blume as α-glucosidase Inhibitor

In vitro and in silico Screening of Bioactive Compounds from Jasminum subtriplinerve Blume as α-glucosidase Inhibitor

Virtual Screening and in Vitro Evaluation to Identify a Potential Xanthine Oxidase Inhibitor Isolated from Vietnamese Uvaria cordata

Xanthine oxidase (XO) is a potential target for gout disease experiments on animals and humans. Using a molecular docking technique to search for anti-XO compounds from Vietnamese medicinal plants, we discovered that numerous compounds from Uvaria cordata (Dunal) Alston (Annonaceae family) showed this activity. Among these, cordauvarin A exhibited the strongest binding affinity (−8.8 kcal/mol) to XO through a binding interaction with 5 amino acids (eg Gln-1194, Ala-1079, Ser-1080, Met-1038, and Arg-912) of XO protein. Lipinski's rule of five was used to predict the druglikeness of this compound. To confirm the inhibitory activity, an in vitro assay was performed, and the results demonstrated that cordauvarin A significantly inhibited XO, with an IC50 of 124.5 ± 10.12 μM. This study reveals that cordauvarin A is a possible natural therapeutic agent for gout treatment and that this genus should be explored more extensively. However, further investigations are necessary to develop possible natural therapeutic medicines for clinical usage.

Cytotoxic constituents from Vietnamese Pterospermum truncatolobatum Gagnep.

Pterospermum truncatolobatum Gagnep. has long been used as a traditional medicine in Vietnam. Its crude extract showed cytotoxicity against human epidermal carcinoma (KB) cell lines. However, its chemical constituent and biological activity remains unknown. In the course of our investigation on the Vietnamese medicinal plants, four compounds, taraxerol ( 1 ), betulonic acid ( 2 ), b -sitosterol ( 3 ) and eicosanoic acid ( 4 ) were purified from methanolic extract of P. truncatolobatum by silica gel column chromatography. Their structures were determined by spectral ( Mass spectrometry and nuclear magnetic resonance ) analysis and by comparison with the literature reports. Of which, betulonic acid ( 2 ) showed moderate cytotoxicity against all four cancer cell lines, KB, MCF7 (human breast carcinoma), LU (human lung carcinoma), and HepG2 (hepatocellular carcinoma).

Essential oils of Uvaria boniana- chemical composition, in vitro bioactivity, docking, and in silico ADMET profiling of selective major compounds.

Phytochemical investigation applying GC (gas chromatography)-MS (mass spectrometry)/GC-FID (flame ionization detection) on the hydro-distilled essential oils of the Vietnamese medicinal plant Uvaria boniana leaf and twig lead to the detection of 35 constituents (97.36%) in the leaf oil and 52 constituents (98.75%) in the twig oil. Monoterpenes, monoterpenoids, sesquiterpenes, and sesquiterpenoids were characteristic of U.boniana essential oils. The leaf oil was represented by major components (E)-caryophyllene (16.90%), bicyclogermacrene (15.95%), α-humulene (14.96%), and linalool (12.40%), whereas four compounds α-cadinol (16.16%), epi-α-muurolol (10.19%), α-pinene (11.01%), and β-pinene (8.08%) were the main ones in the twig oil. As compared with the leaf oil, the twig oil was better in antimicrobial activity. With the same MIC value of 40mg/mL, the twig oil successfully controlled the growth of Gram (+) bacterium Bacillus subtilis, Gram (-) bacterium Escherichia coli, fungus Aspergillus niger, and yeast Saccharomyces cerevisiae. In addition, both two oil samples have induced antiinflammatory activity with the IC50 values of 223.7-240.6mg/mL in NO productive inhibition when BV2 cells had been stimulated by LPS. Docking simulations of four major compounds of U.boniana twig oil on eight relevant antibacterial targets revealed that epi-α-muurolol and α-cadinol are moderate inhibitors of E.coli DNA gyrase subunit B, penicillin binding protein 2X and penicillin binding protein 3 of Pseudomonas aeruginosa with similar free binding energies of-30.1,-29.3, and-29.3kJ/mol, respectively. Furthermore, in silico ADMET studies indicated that all four docked compounds have acceptable oral absorption, low metabolism, and appropriated toxicological profile to be considered further as drug candidates.

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

Ethnopharmacological relevanceCurcuma 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 studyThis 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 methodsThe 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. ResultsGermacrone, 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. ConclusionsOur 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.