• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.163-168
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• 2012

An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

• Korean Journal of Food Science and Technology
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• v.32 no.3
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• pp.706-712
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• 2000

The superoxide dismutase(SOD)-like activities for 26 kinds of herbs and spices and 10 kinds of instant curry products were determined by measuring their abilites to reduce nitroblue tetrazolium. All samples showed the SOD-like activities. Rosemary, cassia, tarragon, allspice, oregano, bay leaves, basil, marjoram, thyme and star anise had higher activities than $10^5\;unit/g$ and clove had highest activity of $232,143{\pm}19.989\;unit/g$. The SOD-like activities for 10 kinds of instant curry products were in the range of $400{\sim}700\;unit/g$ when measured after heat treatment at $100^{\circ}C$ for 10 min. The water extracts of spices, herbs and curries were obtained by heat treatments of $25^{\circ}C$ for 60 min or $100^{\circ}C$ for 10 min, and their nitrite scavenging activity was measured at different pH conditions(1.2, 4.2 or 6.0). The nitrite scavenging activities were higher at acidic pH. However, the effects were not different from two heat treatments. The water extracts from cassia, bay leaves, allspices, oregano, staranise, rosemary, clove and tarragan had high nitrite scavenging activity(>90%) when they were measured at pH 1.2, and those from clove was highest $(97.58{\pm}0.88%)$. The pure curry used as raw materials for instant curry products had the nitrite scavenging activity in the range of $50{\sim}60%$ at pH 1.2 and the activity was not changed during the aging period$(0{\sim}12weeks)$. The ten brands of instant curry products had the nitrite scavenging activities of $12{\sim}28%$ at pH 1.2

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.886-890
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• 2017

In this study, the protective effect of rice with Phellinus linteus mycelium (PLMR) against hydrogen peroxide-induced oxidative stress was assessed in a mouse hippocampal neuronal HT22 cell line through (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) salt (MTS) assay and western blot. MTS assay using HT22 cells showed that PLMR extract did not affect viability at a concentration range from 1 mg/mL to 5 mg/mL. However, at concentrations over 10 mg/mL, PLMR extract resulted in increased cell death. Cell viability of HT22 was significantly reduced by $H_2O_2$ treatment, and reduction of cell viability was efficiently restored by treatment with PLMR extract in a dose-dependent manner from 0.1 to 1 mg/mL. Cells treated with $H_2O_2$ showed increased expression of Bax, a pro-apoptotic protein, which was down-regulated by treatment with PLMR extract. On the other hand, cells treated with $H_2O_2$ resulted in reduced expression of Bcl-2, an anti-apoptotic protein, which was restored by treatment with PLMR extract. In addition, treatment with PLMR extract reduced expression of cleaved caspase 3 and PARP, which were up-regulated by $H_2O_2$ treatment. The results may suggest that treatment with PLMR extract would suppress $H_2O_2$-induced apoptosis of HT22 cells.

• Advances in Traditional Medicine
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• v.5 no.3
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• pp.216-230
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• 2005

Although the field of study in immune enhancing compounds is relatively new, natural products from plants represent a rich and promising source of novel molecules with immunomodulating properties, Microglial cells, the main immune effector cells of the brain, usually display a ramified morphology and low expression levels of immunologically relevant antigens such as MHC class I and class II. Since any compound which participates in activation of phagocytic cells contributes to the production of potentially toxic factors, the search for convenient in vitro test-systems and study of mechanisms of action of these agents are of great interest. Human blood polymorphonuclear (PMN) cells and primary microglial cells isolated from Sprague-Dawley rats were used as cellular screening tests for study of phagocytosis-stimulating action of immunomodulating agents. Numbers of phagocytic activity were evaluated by the phagocyte ingestion of yeast cells and NO-synthase activity, nitrite production, and nitroblue tetrazolium test were determined after phagocyte stimulation. It was possible to demonstrate that indexes of phagocytic activity can be used as quantitative indicators for measurement immunomodulating activity. As a positive control, Zymosan A-induced phagocytosis in both PMN cells and primary microglial cells was used. $IFN-{\gamma}$ (0.1 -1 U/ml) stimulated phagocytosis in PMN cells 1.2 times after 2 - 3 h incubation, although at higher concentrations (10 - 100 U/ml) it strongly inhibited phagocytosis. In a similar way, at higher concentrations, $IFN-{\gamma}$ (100 - 500 U/ml) suppressed phagocytosis in zymosan-A stimulated microglial cells. When Polypodium leucotomus, cambricum and vulgare extracts were tested alone, increased levels of phagocytosis were observed in PMN. In addition, microglial cells showed both increased phagocytosis and MHC class-II antigen expressions. Surprisingly, when PMN and microglia were treated with a combination of Polypodium and $IFN-{\gamma}$, phagocytosis was not inhibited. We did not find changes in NO-synthase activity and nitrite production in both microglia and PMN cells activated by different immunomodulating agents. These results indicate that primary microglial cell cultures as well as human PMN cells can provide reproducible quantitative results in screening phagocytic activity of different immunoactive compounds. Furthermore, both inhibitory or activation mechanisms might be studied using these in vitro experimental approaches.

• Korean Journal of Plant Resources
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• v.24 no.1
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• pp.69-75
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• 2011

This study was conducted to investigate the variation of cone, seed and germination characteristics among populations and among individuals within populations of Picea jezoensis(Siebold & Zuccarini) Carriere distributed in Korea. Cone collected from 25 trees in two natural populations(Mt. Jiri and Mt. Dokyu) and their thirteen cone and seed characteristics as well as three germination behaviors were analyzed. Statistical analysis showed significant differences among populations and among individuals within populations in 9 traits except for seed breadth, seed weight, index of seed wing and mean germination time. Generally, morphological variation of cone and seed wings from Mt. Jiri population has smaller and longer than those of Mt. Dokyu population. Percentage of sound seeds and Tetrazolium test from Mt. Jiri population showed 1.79 and 1.87 higher values than Mt. Dokyu population, respectively. The maximum germination percentage was obtained at the optimum temperature of $20^{\circ}C$ and in this case, mean germination time and germination rate showed 7.5 days and 2.9 ea./day, respectively. In seed germination behaviors, percentage(40.7%) and rate(0.90 ea./day) of germination in Mt. Jiri population were more higher and faster than those of Mt. Dokyu population(17.7%, 0.37 ea./day). According to correlation analysis, P. jezoensis populations with small restricted distribution may have been reduced because seed qualities were correlated with increased levels of inbreeding and disproportion flowering.

• Journal of Korean Medicine for Obesity Research
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• v.19 no.2
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

• Journal of Sasang Constitutional Medicine
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• v.15 no.1
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• pp.72-89
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• 2003

To elucidate the neuroprotective effect of Yeoldahanso-tang(YHT) on nerve cells damaged by hypoxia, the cytotoxic effects of exposure to hypoxia were determined by XTT(SODIUM3,3'-{I-[(PHENYLAMINO) CARBONYL]-3,4-TETRAZOLIUM}- BIS (4-METHOXY-6-NITRO) BENZENE SULFONIC ACID HYDRATE), NR(Neutral red), MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and SRB(Sulforhodamin B) asssay. The activity of catalase and SOD(Superoxide dismutase) was measured by spectrophometry, and $TNF-{\alpha}$(Tumor cell necrosis $fector-{\alpha}$) and PKC(Protein kinase C) activity was measured after exposure to hypoxia and treatment of YHTWE. Also the neuroprotective effect of YHTWE was researched for the elucidatioion of neuroprotective mechanism. The results were as follows; 1. Hypoxia decreased cell viability measured by XTT, NR assay when cultured cerebral neurons were exposed to 95% N2/5% CO2 for $2{\sim}26$ minutes in these cultures and YHTWE inhibited the decrease of cell viability. 2. H2O2 treatment decreased cell viability measured by MTT, and SRB assay when cultured cerebral neurons were exposed to 1-80 ${\mu}M$ for 6 hours, but YHTWE inhibited the decrease of cell viability. 3. Hypoxia decreased catalase and SOD activity, and also $TNF-{\alpha}$ and PKC activity in these cultured cerebral neurons, but YHTWE inhibited the decrease of the catalase and SOD activity in these cultures. 4. Hypoxia triggered the apoptosis via caspase activation and internucleosomal DNA fragmentation. Also hypoxia stimulate the release of cytochrome c forom mitochondria. YHTWE inhibited the apoptosis via caspase activation induced by hypoxia. From these results, it can be suggested that brain ischemia model induced hypoxia showed neurotoxicity on cultured mouse cerebral neurons, and the YHTWE has the neuroprotective effect in blocking the neurotoxicity induced by hypoxia in cultured mouse cerebral neurons.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.40 no.5
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• pp.300-307
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• 2007

This study investigated the effects of dietary supplementation with hot water extracts (HE) of selfheal (Prunella vulgaris) and Lactobacillus rhamnosus culture fluid (LF) on the growth and non-specific immune responses of juvenile olive flounder. A total of 270 fish ($5.07{\pm}0.01g$, average $weight{\pm}SD$) were divided randomly into nine groups, and three groups were fed one of three isonitrogenous (51% crude protein) and isocaloric (17.6 MJ/kg) diets. The diets contained no supplement, 50 mL hot water extract, or 50 mL L. rhamnosus fluid (designated as diets CON, HE and LF, respectively). During the 8-week feeding trial, the growth, feed utilization and survival of the fish were not significantly affected by the experimental diets. There were no significant differences in the hematocrit and hemoglobin of fish fed each of the experimental diets. However, the serum alanine aminotransferase and aspartate aminotransferase activity were lower with the dietary supplements containing HE and LF. The alanine aminotransferase activity of fish fed the HE diet was significantly lower than that of fish fed the control diet. The hematocrit, hemoglobin, and nitroblue tetrazolium (NBT) activity were higher 3 hr after feeding than 24 hr afterwards. The NBT activity of fish fed the HE and LF diets were significantly higher than that of fish fed the control diet 3 hr after feeding. The findings suggest that dietary supplementation with HE and LF could enhance the nonspecific immune responses of juvenile olive flounder.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.562-570
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• 2018

Background: Lung cancer is the leading cause of cancer-related death worldwide. In this study, we used a bioactivity-guided isolation technique to identify constituents of Korean Red Ginseng (KRG) with antiproliferative activity against human lung adenocarcinoma cells. Methods: Bioactivity-guided fractionation and preparative/semipreparative HPLC purification were used with LC/MS analysis to separate the bioactive constituents. Cell viability and apoptosis in human lung cancer cell lines (A549, H1264, H1299, and Calu-6) after treatment with KRG extract fractions and constituents thereof were assessed using the water-soluble tetrazolium salt (WST-1) assay and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, respectively. Caspase activation was assessed by detecting its surrogate marker, cleaved poly adenosine diphosphate (ADP-ribose) polymerase, using an immunoblot assay. The expression and subcellular localization of apoptosis-inducing factor were assessed using immunoblotting and immunofluorescence, respectively. Results and conclusion: Bioactivity-guided fractionation of the KRG extract revealed that its ethyl acetate-soluble fraction exerts significant cytotoxic activity against all human lung cancer cell lines tested by inducing apoptosis. Chemical investigation of the ethyl acetatesoluble fraction led to the isolation of six ginsenosides, including ginsenoside Rb1 (1), ginsenoside Rb2 (2), ginsenoside Rc (3), ginsenoside Rd (4), ginsenoside Rg1 (5), and ginsenoside Rg3 (6). Among the isolated ginsenosides, ginsenoside Rg3 exhibited the most cytotoxic activity against all human lung cancer cell lines examined, with $IC_{50}$ values ranging from $161.1{\mu}M$ to $264.6{\mu}M$. The cytotoxicity of ginsenoside Rg3 was found to be mediated by induction of apoptosis in a caspase-independent manner. These findings provide experimental evidence for a novel biological activity of ginsenoside Rg3 against human lung cancer cells.

• Korean Journal of Plant Resources
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• v.30 no.1
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• pp.50-57
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• 2017

This study aimed to investigate the morphological characteristics and optimal germination conditions in seeds of Arabis pendula L., a traditionally edible and medicinal plant. The external seed shape was circular-obovate with narrow wings and dark brown. The seed length and width were 1.25 mm and 1.47 mm, respectively. The seeds were exalbuminous and the embryo was a bent type. Seed germination was the highest (49.7%) at $20^{\circ}C$ under dark conditions among the various temperature and light conditions applied. However, under the dark condition, the seedling was weak, overgrown, and the cotyledons were small and folded. To improve the germination and growth of seedlings, the seeds were pre-treated with $GA_3$ solutions of varying concentrations (0, 200, 500, and 1,000 mg/L). The seed germination and seedling growth were effectively improved by $GA_3$ pre-treatment. The germination rate was the highest (97.3%), mean germination time was the shortest (8.1 days), and a vigorous growth of seedlings was observed upon pre-soaking the seeds in 500 mg/L $GA_3$ solution. In conclusion, the best method for germination was pre-soaking in 500 mg/L $GA_3$ ($4^{\circ}C$, dark, 24 h) and incubating the seeds at $20^{\circ}C$ for 15 days.