Browsing by Author "Tamtin M."

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    Acceleration of Ovarian Maturation in the Female Mud Crab With RNA Interference of the Vitellogenesis-Inhibiting Hormone (VIH)
    (2022-06-02) Duangprom S.; Saetan J.; Phanaksri T.; Songkoomkrong S.; Surinlert P.; Tamtin M.; Sobhon P.; Kornthong N.; Mahidol University
    In the present study, double strand RNA technology (dsRNA) was used to inhibit transcripts of vitellogenesis-inhibiting hormone (VIH) that mainly synthesized and secreted from the central nervous system in Scylla olivacea females. The results presented in this study clearly demonstrate the potential dsRNA-VIH was highly effective to inhibit VIH in the eyestalks of females injected with dsRNA-VIH on the 3rd, 7th and 14th day, respectively. The dsRNA-VIH injections were performed at 14-day intervals, a single dsRNA dose of 0.6 µg/gram body weight was enough to suppress VIH expression until 14th day after injection. The dsRNA-VIH injection significantly increased gonad-somatic index (GSI) and hemolymph vitellin level at day 14 and 28 when compared with control groups. The histological observation found that the number of oocyte step 4 in dsRNA-VIH group was significantly higher than that of the control group. Also, dsRNA-VIH has stimulatory function on other reproduction-related genes such as the Scyol-PGES and Scyol-ESULT that both genes gradually increased their expressions in brain and ventral nerve cord. In conclusion, the silence of VIH gene could reduce the production of VIH from eyestalk and brain that affected other downstream genes related to ovarian maturation in the mud crab.
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    Anti-Obesity Effects of Marine Macroalgae Extract Caulerpa lentillifera in a Caenorhabditis elegans Model
    (2023-11-03) Chumphoochai K.; Manohong P.; Niamnont N.; Tamtin M.; Sobhon P.; Meemon K.; Mahidol University
    Obesity is a multifactorial disease characterized by an excessive accumulation of fat, which in turn poses a significant risk to health. Bioactive compounds obtained from macroalgae have demonstrated their efficacy in combating obesity in various animal models. The green macroalgae Caulerpa lentillifera (CL) contains numerous active constituents. Hence, in the present study, we aimed to elucidate the beneficial anti-obesity effects of extracts derived from C. lentillifera using a Caenorhabditis elegans obesity model. The ethanol (CLET) and ethyl acetate (CLEA) extracts caused a significant decrease in fat consumption, reaching up to approximately 50-60%. Triglyceride levels in 50 mM glucose-fed worms were significantly reduced by approximately 200%. The GFP-labeled dhs-3, a marker for lipid droplets, exhibited a significant reduction in its level to approximately 30%. Furthermore, the level of intracellular ROS displayed a significant decrease of 18.26 to 23.91% in high-glucose-fed worms treated with CL extracts, while their lifespan remained unchanged. Additionally, the mRNA expression of genes associated with lipogenesis, such as sbp-1, showed a significant down-regulation following treatment with CL extracts. This finding was supported by a significant decrease (at 16.22-18.29%) in GFP-labeled sbp-1 gene expression. These results suggest that C. lentillifera extracts may facilitate a reduction in total fat accumulation induced by glucose through sbp-1 pathways. In summary, this study highlights the anti-obesity potential of compounds derived from C. lentillifera extracts in a C. elegans model of obesity, mediated by the suppression of lipogenesis pathways.
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    Apoptotic and Autophagic Cell Death Effects of the Hexane Extract of Tropical Marine Algae Halymenia durvillei against Human Glioblastoma Cells: In vitro and in silico Studies
    (2024-02-01) Vivithanaporn P.; Siangcham T.; Tanawoot V.; Settacomkul R.; Pranweerapaiboon K.; Meemon K.; Niamnont N.; Tamtin M.; Sobhon P.; Sangpairoj K.; Vivithanaporn P.; Mahidol University
    Glioblastoma (GBM) considered as aggressive brain cancer with high mortality rate in patients even after surgical resection. Resistant to chemotherapy is the major problem in GBM therapy. Discovery of novel bioactive compounds from algae is being investigated as alternative sources for potential treatment as well as prevention in glioblastoma. This study revealed the effects of marine red algae extract from hexane solvent fraction of Halymenia durvillei (HDHE) on proliferation and cell death in A172 human GBM cells. HDHE decreased proliferation and promoted cell cycle arrest at G2/M phase. HDHE induced apoptotic cell death in A172 cells through mitochondrial membrane dysfunction, the decrease of anti-apoptotic Bcl-2 protein expression, and activation of caspase 3/7. Moreover, HDHE increased intracellular reactive oxygen species (ROS) production and accumulation of LC3-II, an autophagic marker. The docked conformation of palmitic acid, a major component of HDHE, showed a high affinity binding to TP53 and Beclin-1 as cell death-related target molecules. This research conclusively demonstrated that HDHE might serve as a potent anticancer agent against glioblastoma by promoting apoptotic and autophagic cell death in A172 human GBM cells.
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    Ethyl Acetate Extract of Halymenia durvillei Induced Apoptosis, Autophagy, and Cell Cycle Arrest in Colorectal Cancer Cells
    (2023-03-01) Chantree P.; Martviset P.; Sornchuer P.; Thongsepee N.; Sangpairoj K.; Meemon K.; Niamnont N.; Tamtin M.; Sobhon P.; Mahidol University
    Colorectal cancer is one of the most death-dealing cancers. However, conventional cancer treatments still have side effects. Therefore, novel chemotherapeutic agents with less side effects are still in search. A marine red seaweed, Halymenia durvillei, is recently interested in its anticancer effects. This study investigated the anticancer effect of ethyl acetate extract of H. durvillei (HDEA) on HT-29 colorectal cancer cells in association with the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. HDEA-treated HT-29 and OUMS-36 cells were used for cell viability tests by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide assay. The effects of HDEA on apoptosis and cell cycle were evaluated. The nuclear morphology and mitochondrial membrane potential (∆ψm) were observed by Hoechst 33342 and JC-1 staining, respectively. The gene expression of PI3K, AKT, and mTOR genes was evaluated using a real-time semiquantitative reverse transcription-polymerase chain reaction. The corresponding protein expressions were assessed by western blot analysis. The result revealed that the cell viability of treated HT-29 cells diminished while that of OUMS-36 cells was non-significant. By the down-regulation of cyclin-dependent kinase 4 and cyclin D1, HDEA-treated HT-29 cells were arrested in the G0/G1 phase. By the up-regulation of cleaved poly(adenosine diphosphate-ribose) polymerase, caspase-9, caspase-8, caspase-3, and Bax, HDEA-treated HT-29 cells underwent apoptosis, but suppressed Bcl-2, disrupted nuclear morphology and ∆ψm. Furthermore, treated HT-29 cells underwent autophagy by up-regulation of light chain 3-II and beclin-1. Lastly, HDEA suppressed the expression of PI3K, AKT, and mTOR. Therefore, HDEA exerts anticancer effects against HT-29 cells, confirmed by apoptosis, autophagy, and cell cycle arrest induction via regulation of the PI3K/AKT/mTOR signaling pathway.
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    Ethyl Acetate Extract of Marine Algae, Halymenia durvillei, Provides Photoprotection against UV-Exposure in L929 and HaCaT Cells
    (2022-11-11) Kraokaew P.; Manohong P.; Prasertsuksri P.; Jattujan P.; Niamnont N.; Tamtin M.; Sobhon P.; Meemon K.; Mahidol University
    Halymenia durvillei is a red alga distributed along the coasts of Southeast Asian countries including Thailand. Previous studies have shown that an ethyl acetate fraction of H. durvillei (HDEA), containing major compounds including n-hexadecanoic acid, 2-butyl-5-hexyloctahydro-1H-indene, 3-(hydroxyacetyl) indole and indole-3-carboxylic acid, possesses high antioxidant and anti-lung cancer activities. The present study demonstrated that HDEA could protect mouse skin fibroblasts (L929) and human immortalized keratinocytes (HaCaT) against photoaging due to ultraviolet A and B (UVA and UVB) by reducing intracellular reactive oxygen species (ROS) and expressions of matrix metalloproteinases (MMP1 and MMP3), as well as increasing Nrf2 nuclear translocation, upregulations of mRNA transcripts of antioxidant enzymes, including superoxide dismutase (SOD), heme oxygenase (HMOX) and glutathione S-transferase pi1 (GSTP1), and procollagen synthesis. The results indicate that HDEA has the potential to protect skin cells from UV irradiation through the activation of the Nrf2 pathway, which leads to decreasing intracellular ROS and MMP production, along with the restoration of skin collagen.
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    Evaluation of Toxicity and Anti-Oxidation Activity of the Extracts from Halymenia durvillei
    (2022-03-15) Chaiwichien A.; Samrit T.; Osotprasit S.; Kueakhai P.; Sobhon P.; Meemon K.; Niamnont N.; Manohong P.; Pranweerapaiboon K.; Tamtin M.; Changklungmoa N.; Mahidol University
    Halymenia durvillei (HD), a marine red alga, is believed to have potentials for pharmacological, nutritional and cosmetic applications. However, such potentials are acceptable only when their extracts are devoid of any adverse effects on human health. No previous research has been conducted the toxicity and anti-oxidation capacity of HD. Thus, the aim of this work was to investigate toxicity and anti-oxidation activities of HD extracts. In this study, the toxicity and anti-oxidation capacity of 5 fractions of HD solvent extracts, i.e., ethanol (HDET), hexane (HDHE), ethyl acetate (HDEA), butanol (HDBU), and aqueous (HDAQ) were evaluated. The cytotoxicity was evaluated by MTT and LDH assays on 4 cell types, i.e., fibroblast, macrophage, hepatocyte and keratinocyte. The genotoxicity was evaluated by comet assay and micronucleus test using TK6 lymphoblastoid cell line. The anti-oxidation capacity was investigated by DPPH and ABTS assays. The toxicity studies showed that HDET, HDBU, HDAQ had very low to no toxicity as indicated by cytotoxicity and genotoxicity tests while HDEA, HDHE have some toxicity at high concentrations. HDAQ showed low antioxidant activity while HDET, HDEA, HDHE and HDBU possess relatively high antioxidant activity. Overall, our results indicated that HDET and HDAQ could be consumed as they are not toxic and HDHE, HDEA, and HDBU could be safely consumed at doses lower than 100 μg/mL. Further investigation using in vivo assays are needed to ensure the safety of HD extracts for animal and human consumptions.
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    Evaluation of Toxicity and Antioxidant Activity of the Ethanolic Extract from Ulva Lactuca
    (2025-02-01) Osotprasit S.; Samrit T.; Savedvanich G.; Chaiwichien A.; Changklungmoa N.; Kueakhai P.; Athipornchai A.; Tamtin M.; Sobhon P.; Jaikua W.; Osotprasit S.; Mahidol University
    Ulva lactuca (sea lettuce) is a popular food for consumers and has high nutritional value. It has also been found to be a source of health-promoting substances, including a high amount of proteins which can be utilized as a future plant-based functional ingredient in the food industry. However, before extensive applications in the aforementioned fields, it is crucial to ensure the extracts’ biological-pharmacological activities and their safety. In this study, we evaluated the cytotoxicity, genotoxicity, and antioxidant activity of the ethanolic extract of the sea lettuce extract.We tested cytotoxicity by MTT assays on 5 cell lines, including fibroblast (L929), macrophages (RAW 264.7), hepatocytes (FL83B), keratinocytes (HaCaT) and normal colon (CCD-18co) cells. Genotoxicity of the extract was tested by comet and micronucleus assays on human lymphoblast cells (TK6). Our results demonstrated at a concentration lower than 50 µg/mL the extract did not show any cytotoxicity as well as genotoxicity. The extract was shown to possess high antioxidant capacity by DPPH and ABTS scavenging at the EC50 value of U. lactuca extracts was 631.84 ± 5.64 and 330.35 ± 19.49 μg/mL, respectively. Therefore, the extract may be safely used as an ingredient for food, that should help to ensure food security for humans in the future, as well as for cosmetic, pharmaceutical and other health-promoting products.
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    Extracts of tropical green seaweed Caulerpa lentillifera reduce hepatic lipid accumulation by modulating lipid metabolism molecules in HepG2 cells
    (2024-03-30) Sangpairoj K.; Pranweerapaiboon K.; Saengkhae C.; Meemon K.; Niamnont N.; Tamtin M.; Sobhon P.; Yisarakun W.; Siangcham T.; Sangpairoj K.; Mahidol University
    Seaweed has attracted attention as a bioactive source for preventing different chronic diseases, including liver injury and non-alcoholic fatty liver disease, the leading cause of liver-related mortality. Caulerpa lentillifera is characterized as tropical edible seaweed, currently being investigated for health benefits of its extracts and bioactive substances. This study examined the effects of C. lentillifera extract in ethyl acetate fraction (CLEA) on controlling lipid accumulation and lipid metabolism in HepG2 cells induced with oleic acid through the in vitro hepatic steatosis model. Gas chromatography-mass spectrometry (GC-MS) analysis indicated that CLEA contained diverse organic compounds, including hydrocarbons, amino acids, and carboxylic acids. Docked conformation of dl-2-phenyltryptophane and benzoic acid, two major bioactive CLEA components, showed high affinity binding to SIRT1 and AMPK as target molecules of lipid metabolism. CLEA reduced lipid accumulation and intracellular triglyceride levels in HepG2 cells stimulated with oleic acid. The effect of CLEA on regulating expression of lipid metabolism-related molecules was investigated by qPCR and immunoblotting. CLEA promoted expression of the SIRT1 gene in oleic acid-treated HepG2 cells. CLEA also reduced expression levels of SREBF1, FAS, and ACC genes, which might be related to activation of AMPK signaling in lipid-accumulated HepG2 cells. These findings suggest that CLEA contains bioactive compounds potentially reducing triglyceride accumulation in lipid-accumulated HepG2 hepatocytes by controlling lipid metabolism molecules.
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    Hexadecanoic acid-enriched extract of Halymenia durvillei induces apoptotic and autophagic death of human triple-negative breast cancer cells by upregulating ER stress
    (2022-03-01) Sangpairoj K.; Settacomkul R.; Siangcham T.; Meemon K.; Niamnont N.; Sornkaew N.; Tamtin M.; Sobhon P.; Vivithanaporn P.; Mahidol University
    Objective: To investigate the effect of the hexane solvent fraction of Halymenia durvillei (HDHE) on triple-negative breast cancer. Methods: The phytochemical profile of HDHE was investigated by GC-MS. The cytotoxicity of HDHE against MDA-MB-231 cells was determined. The apoptotic and autophagic effects of HDHE were analyzed. The expression of molecular markers controlling apoptosis, autophagy, DNA damage, and endoplasmic reticulum (ER) stress was determined. Results: HDHE contains a mixture of fatty acids, mainly hexadecanoic acid. HDHE at a cytotoxic concentration induced apoptotic death of MDA-MB-231 cells through mitochondrial membrane dysfunction, and induction of apoptosis markers, and increased the expression of proteins related to DNA damage response. HDHE also induced the expression of LC-3, a marker of autophagic cell death at a cytotoxic concentration. Moreover, HDHE modulated the expression of ER stress genes. Conclusions: The hexadecanoic acid-enriched extract of Halymenia durvillei promotes apoptosis and autophagy of human triple-negative breast cancer cells. This extract may be further explored as an anticancer agent for triple-negative breast cancer.
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    Potent ovarian development as being stimulated by cocktail hormone in the female Scylla olivacea
    (2023-01-01) Saetan J.; Duangprom S.; Songkoomkrong S.; Amonruttanapun P.; Phanaksri T.; Surinlert P.; Samhuay C.; Tamtin M.; Suwansa-Ard S.; Cummins S.F.; Sobhon P.; Kornthong N.; Saetan J.; Mahidol University
    The mud crab Scylla olivacea is widely cultured for its economic value, but reproduction issues limit its production. Vitellogenesis-inhibiting hormone (VIH), serotonin (5-HT), and gonadotropin-releasing hormone (GnRH) are important neurohormones that control reproduction in crustaceans. Mimicking crab hormone stimulation during reproduction has scarcely been reported. Comparison of the single hormone and multiple hormone approaches to crab hormonal control in S. olivacea is limited. In situ hybridization showed that injection of dsRNA-VIH could abolish its gene expression in neuronal clusters of female S. olivacea eyestalks, potentially reducing its inhibitory effects on ovarian maturation. This was confirmed by assessing the ovarian gonadosomatic index (GSI), hemolymph vitellogenin (Vg), an indicator of vitellogenesis, and gonad histology using dsRNA-VIH and 5-HT/GnRH combinations. Based on our findings, we demonstrated that administration of dsRNA-VIH significantly increased the gonadosomatic index (GSI) on days 14 and 28 post-treatment. The combination cocktail, however, consisting of 5-HT + GnRH + dsRNA-VIH on days 14 and 28, and GnRH + dsRNA-VIH on day 28, was the most efficacious in increasing GSI and enhancing crab ovarian maturation. Upregulation of hemolymph Vg levels was observed solely on the 28th day following treatment with dsRNA-VIH, 5-HT + GnRH + dsRNA-VIH, and GnRH + dsRNA-VIH. Differential gene expression analysis using quantitative RNA-sequencing of the neural tissues (brain and ventral nerve cord), revealed a significant upregulation of certain receptors (5-HTR, GnRHR, LHR, and FSHR), neuropeptides (sNPF, NPF1, NPF2, SIFamide, AKH/Crz, CHH, and RPCH), downstream reproductive-related genes (FAMeT, ESULT, progesterone-like protein), and prostanoid-related genes (phospholipase A and C, COX, Thromboxane A synthase, prostaglandin D, E, and F synthases) following treatment, particularly dsRNA-VIH + GnRH and/or 5-HT-injected individuals. Upregulation of prostaglandin E synthase and estrogen sulfotransferase genes was confirmed by real-time PCR. Since the construction and propagation of dsRNA-VIH is costly, its lower dose application supplemented with synthetic GnRH and/or 5-HT may be an alternative approach to ensure that female S. olivacea attain sufficient reproductive fecundity in aquaculture. Furthermore, we propose that the administration of multiple hormones in crabs may better emulate the physiological conditions of crustaceans in their natural habitat.
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    The occurrence of luteinizing hormone-like molecule and its receptor in the blue swimming crab, Portunus pelagicus
    (2025-01-01) Saetan U.; Kornthong N.; Duangprom S.; Songkoomkrong S.; Phanthong P.; Sanprick A.; Tipbunjong C.; Tamtin M.; Saetan J.; Saetan U.; Mahidol University
    Knowledge of the neuroendocrine system possibly improves the reproductive performance of captivated crustacean broodstock in aquaculture and it may substitute eyestalk ablation. In this study, we explored the luteinizing hormone (LH)-like molecule and proved the existence of the LH receptor (PpelLHR)-like mRNA in the blue swimming crab, Portunus pelagicus. Using the anti-human LH-β antibody, the immunoreactivities were found in the central nervous system (CNS) and ovary of the crab with the strongest signal in the mature ovary. The full-length PpelLHR-like mRNA sequence contained 4818 bp with deduced protein predicted as seven transmembrane G-protein coupled receptor, made of 1605 amino acids. The phylogenetic tree suggested this protein belonged to the clade of invertebrate LHR/FSHR-like proteins. The PpelLHR-like mRNA expressed in various organs and real-time qPCR revealed significantly higher expression of this mRNA in the brain and lower expression in the ovary of the mature crabs. In situ hybridization of this mRNA was demonstrated in neuronal clusters of the brain, ventral nerve cord, and in the oocyte stage 1–4 of the ovary, respectively. This study was preliminary to prove the existence of LH and its receptor in the blue swimming crab. Functional assay of this receptor should be performed as the next part of experiments to firmly conclude its appearance.
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    The Scavenging Activity and Safety Effect of Red Marine Algae Acanthophora spicifera Ethanol Extract
    (2024-01-01) Samrit T.; Osodprasit S.; Chaiwichien A.; Savedvanich G.; Changklungmoa N.; Kueakhai P.; Athipornchai A.; Tamtin M.; Sobhon P.; Jaikua W.; Samrit T.; Mahidol University
    The red algae, Acanthophora spicifera, are major species grown in Asia and Pacific areas, especially along the coast of southern Thailand. They have the fast growth rate that leads to an abundant supply; thus, they are currently used for wastewater treatment. Furthermore, an ethanolic extract from A. spicifera has been used as a food supplement because of its perceived antioxidant property. Therefore, this study aims to ascertain and evaluate the antioxidant activity and safety of the ethanolic extract from A. spicifera. Safety tests showed that the extract did not have cytotoxicity and genotoxicity effects at concentrations less than 100 µg/mL. In term of antioxidant activity, the DPPH and ABTS assays showed a Trolox equivalent antioxidant capacity (TEAC) of 5.24±0.44 and 4.03±0.13 mg Trolox/g of the extract, respectively. In conclusion, A. spicifera extract is relatively safe and has a rather high efficacy in scavenging free radicals, which makes it suitable for use as food supplement and cosmetics. However, there are a number of caveats regarding its uses in terms of excessive concentrations and exposure duration to avoid causing cytotoxicity and genotoxicity.