Polyenylphosphatidylcholine (PPC), a subgroup of the bioactive agents in phosphatidylcholine (PC), has been indicated to possess liver-protective effects. This study aimed to investigate a promising ...and feasible method to determine PC molecular species with a reverse phase (RP) high-performance liquid chromatograph (HPLC) equipped with an evaporative light scattering detector (ELSD). Chromatography was achieved using a C30 column and an isocratic mobile phase consisting of acetonitrile/methanol/triethylamine (40/58/2, v/v/v) at a flow rate of 1 mL/min, and ELSD detection was performed using 80 °C for the drift tube and an air flow rate of 1.8 L/min. To identify individual peaks on the chromatogram, MALDI-TOF-MS was employed for initial detection, and then the results were used to investigate the relationship between the retention time and fatty acyl chains of each PC molecule. A linear correlation was observed between the retention time and theoretical carbon number (TCN) of individual PC species. The compositions of PC molecular species in soybean and sunflower lecithins were similar to each other, and the major PC molecular species were 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (LLPC), 1-oleoyl-2-linoleoyl-sn-glycero-3-phosphocholine (OLPC), and 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC). The contents of LLPC in soybean PC and sunflower PC were 40.6% and 64.3%, respectively.
Sinularin is an active compound isolated from the cultured soft coral Sinularia flexibilis. In this study, we investigated the effects of sinularin on two human gastric cancer cell lines, AGS and ...NCI-N87. Our results demonstrated that sinularin suppressed the proliferation of gastric cancer cells in a dose-dependent manner and induced apoptosis. In addition, the loss of mitochondrial membrane potential, the release of cytochrome C, the activation of Bax, Bad and caspase-3/9, and the suppression of p-Bad, Bcl-xL and Bcl-2 were observed in the cells treated with sinularin. This finding suggests that sinularin-induced apoptosis is associated with mitochondria-mediated apoptosis and occurs through caspase-dependent pathways. Furthermore, sinularin inhibited the phosphoinositol 3-kinase/Akt/mechanistic target of the rapamycin signaling pathway. Taken together, our results show that sinularin-induced apoptosis is mediated by activation of the caspase cascade and mitochondrial dysfunction. Our findings suggest that sinularin merits further evaluation as a chemotherapeutic agent for human gastric cancer.
Sinulariolide, an active compound isolated from the cultured soft coral Sinularia flexibilis, has potent anti-microbial and anti-tumorigenesis effects towards melanoma and bladder cancer cells. In ...this study, we investigated the effects of sinulariolide on hepatocellular carcinoma (HCC) cell growth and protein expression. Sinulariolide suppressed the proliferation and colony formation of HCC HA22T cells in a dose-dependent manner and induced both early and late apoptosis according to flow cytometry, Annexin V/PI stain and TUNEL/DAPI stain analyses. A mechanistic analysis demonstrated that sinulariolide-induced apoptosis was activated through a mitochondria-related pathway, showing up-regulation of Bax, Bad and AIF, and down- regulation of Bcl-2, Bcl-xL, MCl-1 and p-Bad. Sinulariolide treatment led to loss of the mitochondrial membrane potential, release of mitochondrial cytochrome c to the cytosol, and activation of both caspase-9 and caspase-3. Sinulariolide-induced apoptosis was significantly blocked by the caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK. The increased expression of cleaved PARP also suggested that caspase-independent apoptotic pathway was involved. In the western blotting; the elevation of ER chaperones GRP78; GRP94; and CALR; as well as up-regulations of PERK/eIF2α/ATF4/CHOP; and diminished cell death with pre-treatment of eIF2α phosphatase inhibitor; salubrinal; implicated the involvement of ER stress-mediated PERK/eIF2α/ATF4/CHOP apoptotic pathway following sinulariolide treatment in hepatoma cells. The current study suggested sinulariolide-induced hepatoma cell cytotoxicity involved multiple apoptotic signal pathways. This may implicate that sinulariolide is a potential compound for the treatment of hepatocellular carcinoma.
Sinulariolide, an isolated compound from the soft coral Sinularia flexibilis, possesses the anti-proliferative, anti-migratory and apoptosis-inducing activities against the TSGH bladder carcinoma ...cell. The anti-tumor effects of sinulariolide were determined by 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, cell migration assay and flow cytometry, respectively. Sinulariolide inhibited the growth and migration of bladder carcinoma cells in a dose-dependent manner, as well as induced both early and late apoptosis as determined by the flow cytometer. Also, the sinulariolide-induced apoptosis is related to the mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by the loss of mitochondrial membrane potential, release of cytochrome C, activation of caspase-3/-9, Bax and Bad, as well as suppression of Bcl-2/Bcl-xL/Mcl-1. Detection of the PARP-1 cleaved product suggested the partial involvement of caspase-independent pathways. Moreover, inhibition of p38MAPK activity leads to the rescue of the cell cytotoxicity of sinulariolide-treated TSGH cells, indicating that the p38MAPK pathway is also involved in the sinulariolide-induced cell apoptosis. Altogether, these results suggest that sinulariolide induces apoptosis against bladder cancer cells through mitochondrial-related and p38MAPK pathways.
Safrole is a well-known carcinogenic agent that is present in camphor trees. In this study, a gas chromatographic method was established to quantitate the levels of safrole in essential oils using ...n-decyl alcohol as an internal standard. The method used a nonpolar column and was able to detect concentrations of safrole as low as 5 µg/ml in the samples. Following addition of 2-10 mg of safrole into 1 g of essential oil extracted from Stout Camphor wood (
Hayata) or 1-10 mg of safrole into 1 g of essential oil extracted from Small-flower Camphor wood (
Hayat), the recovery rates of safrole were determined. With direct injection of samples into the gas chromatograph, the results showed that the recovery was more than 96.1%, with a coefficient of variation below 5.6%. We then analyzed 23 commercially available Stout Camphor and other essential oil samples and found that 21 of them contained safrole in the range of 37.65-355.07 mg/g. In addition, in the heavier essential oil distilled from Small-flower Camphor wood, the safrole level was up to 642.98 mg/g. Our results demonstrated that most camphor essential oils on the market have a carcinogenic potential due to their high safrole levels.
In this study, quantitative analysis of sclerotiorin from Penicillium sclerotiorum was established by using HPLC/colorimetry combination method at the condition of unavailability of authentic ...sclerotiorin compound. Mycelium was extracted with methanol, then analyzed by HPLC (a C18 reverse phase column with acetonitrile/water = 65/35, v/v as mobile phase and detected at 370 nm) and simultaneously measured OD sub(370) with spectrophotometers. The limit of quantitation (LOQ) was 0.5 mu g/mL. Recovery studies were performed using the lyophilized mycelia, with each spiked with sclerotiorin at 1 and 10 mg, respectively. The recoveries were found in the range of 97-109%. The coefficients of variation were less than 3.8% with this HPLC/OD sub(370) combination method.
In this study, quantitative analysis of sclerotiorin from Penicillium sclerotiorum was established by using HPLC/colorimetry combination method at the condition of unavailability of authentic ...sclerotiorin compound. Mycelium was extracted with methanol, then analyzed by HPLC (a C18 reverse phase column with acetonitrile/water = 65/35, v/v as mobile phase and detected at 370 nm) and simultaneously measured OD370 with spectrophotometers. The limit of quantitation (LOQ) was 0.5 μg/mL. Recovery studies were performed using the lyophilized mycelia, with each spiked with sclerotiorin at 1 and 10 mg, respectively. The recoveries were found in the range of 97~109%. The coefficients of variation were less than 3.8% with this HPLC / OD370 combination method.
博士
國立臺灣大學
農業化學研究所
92
In this study, a fungal strain with high quantity of yellow pigment was isolated from an orchard soil from Pingtung in our laboratory. This strain was identified as Penicillium ...sclerotiorum and was designated as Penicillium sclerotiorum NTU-FC-13. A yellow needle crystal was obtained by silica-gel column chromatography from the crude pigments, and was identified as sclerotiorin through MS, IR, and NMR spectra. The results showed that the potato agar medium was the best for producing pigments among natural mediums. Maltose could significantly increase the pigment production. Neither organic nor inorganic nitrogen source would influence the pigments production but could increase the volume of biomass. Adding 0.1% sodium chloride could promote the productivity. P. sclerotiorum NTU-FC-13 produces sclerotiorin yellow pigment under submerged fermentation in potato maltose medium (15% potato, 2% maltose, 0.1% NaCl ), the sclerotiorin yield is 633 mg L-1 culture fluid and 39 mg g-1dry cell weight
Sinulariolide, an active compound isolated from the cultured soft coral Sinularia flexibilis, has potent anti-microbial and anti-tumorigenesis effects towards melanoma and bladder cancer cells. In ...this study, we investigated the effects of sinulariolide on hepatocellular carcinoma (HCC) cell growth and protein expression. Sinulariolide suppressed the proliferation and colony formation of HCC HA22T cells in a dose-dependent manner and induced both early and late apoptosis according to flow cytometry, Annexin V/PI stain and TUNEL/DAPI stain analyses. A mechanistic analysis demonstrated that sinulariolide-induced apoptosis was activated through a mitochondria-related pathway, showing up-regulation of Bax, Bad and AIF, and down- regulation of Bcl-2, Bcl-xL, MCl-1 and p-Bad. Sinulariolide treatment led to loss of the mitochondrial membrane potential, release of mitochondrial cytochrome c to the cytosol, and activation of both caspase-9 and caspase-3. Sinulariolide-induced apoptosis was significantly blocked by the caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK. The increased expression of cleaved PARP also suggested that caspase-independent apoptotic pathway was involved. In the western blotting; the elevation of ER chaperones GRP78; GRP94; and CALR; as well as up-regulations of PERK/eIF2α/ATF4/CHOP; and diminished cell death with pre-treatment of eIF2α phosphatase inhibitor; salubrinal; implicated the involvement of ER stress-mediated PERK/eIF2α/ATF4/CHOP apoptotic pathway following sinulariolide treatment in hepatoma cells. The current study suggested sinulariolide-induced hepatoma cell cytotoxicity involved multiple apoptotic signal pathways. This may implicate that sinulariolide is a potential compound for the treatment of hepatocellular carcinoma.
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