• 제목/요약/키워드: Tetrazolium Test

검색결과 121건 처리시간 0.16초

Effects of Dietary Hizikia fusiformis on Growth and Immune Responses in Juvenile Olive Flounder (Paralichthys olivaceus)

  • Pham, Minh Anh;Lee, Kyeong-Jun;Lee, Bong-Joo;Lim, Se-Jin;Kim, Sung-Sam;Lee, Young-Don;Heo, Moon-Soo;Lee, Ki-Wan
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제19권12호
    • /
    • pp.1769-1775
    • /
    • 2006
  • An eight week feeding trial was conducted to investigate the effects of dietary supplementation of hizikia (Hizikia fusiformis) on growth performance, immune responses and resistance of juvenile olive flounder (Paralichthys olivaceus) to Streptococcus iniae. Four experimental diets (designated as Hiz 0, Hiz 2, Hiz 4 and Hiz 6) were formulated to be isonitrogenous (50% crude protein) and isocaloric (17.2 MJ/kg DM). Hizikia powder was added at 0%, 2%, 4% and 6% in diets Hiz 0, Hiz 2, Hiz 4 and Hiz 6, respectively. Three replicates of fish groups (15 fish/tank) were fed one of the experimental diets. At the end of feeding trial, no significant differences were observed in final body weight, specific growth rate, protein efficiency ratio, feed utilization and feed intake among fish groups fed the experimental diets. However, there was clear trend that the growth performances of fish were improved by the increment of dietary hizikia showing a positive growth effects. Mean phagocytes activated with nitro-blue-tetrazolium were significantly increased with the increment of dietary hizikia. The cumulative mortality was significantly (p<0.05) lower in the fish groups fed Hiz 6 diet (no mortality) than that in the other fish groups for 15 days of S. iniae challenge test. The findings of this study suggest that a dietary supplementation of hizikia could enhance the nonspecific immune response and improve the resistance of juvenile olive flounder to S. iniae.

인진항암단의 인간 전골수성 백혈병 세포 사멸 효과 (The Effect of Paljinhangahm-dan on Human Promyelocytic Leukemia Cells(HL-60) Death)

  • 이건업;문석재;문구원;원진희;김동웅;김형균;문영호;이종범;윤준철;김흥준
    • 동의생리병리학회지
    • /
    • 제16권3호
    • /
    • pp.514-520
    • /
    • 2002
  • Paljinhangahm-dan is an Oriental herbal formulation for its ability to modulate cancer cell growth and survival. This research was performed to study the anti-cancer effects of Paljinhangahm-dan water extract(PHWE) in human pro myelocytic leukemia(HL-60) cells. After HL-60 cells were routinely cultured, tetrazolium-based colorimetric(MTT) assay was performed for cytotoxicity test. To explore the mechanism of cytotoxicity. I used several measures of apoptosis to determine whether this processes was involved in PHWE-induced cell death in HL-60 cells. In addition, the experiment was practised 1 H-NMR spectroscopy to examine molecular structure of PHWE. This study suggested that PHWE control cancer cell growth through of apoptosis with less cytotoxicity in normal cells.

Methanol Extract of Polygalae Radix Protects Excitotoxicity in Cultured Neuronal Cells

  • Ban, Ju-Yeon;Lee, Hyun-Joo;Lee, Soo-Bae;Lee, Young-Jong;Seong, Nak-Sul;Song, Kyung-Sik;Bae, Ki-Whan;Seong, Yeon-Hee
    • 한국약용작물학회지
    • /
    • 제11권4호
    • /
    • pp.298-305
    • /
    • 2003
  • Polygalae Radix (PR) from Polygala tenuifolia. (Polygalaceae) is traditionally used in China and Korea, since this herb has a sedative, antiinflammatory, and antibacterial agent. To extend pharmacological actions of PR in the CNS on the basis of its CNS inhibitory effect, the present study examined whether PR has the neuroprotective action against kainic acid (KA) -induced cell death in primarily cultured rat cerebellar granule neurons. PR, over a concentration range of 0.05 to $5{\mu}g/ml$ inhibited KA $(500\;{\mu}M)$-induced neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-y1]-2,5-diphenyl-tetrazolium bromide (MTT) assay. PR $(0.5{\mu}g/ml)$ inhibited glutamate release into medium induced by KA $(500\;{\mu}M)$, which was measured by HPLC. Pretreatment of PR $(0.5{\mu}g/ml)$ inhibited KA $(500\;{\mu}M)$-induced elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$ which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that PR prevents KA-induced neuronal cell damage in vitro.

The venom of jellyfish, Chrysaora pacifica, induces neurotoxicity via activating Ca2+-mediated ROS signaling in HT-22 cells

  • Yang, Yoon-Sil;Kang, Young-Joon;Kim, Hye-Ji;Kim, Min-Soo;Jung, Sung-Cherl
    • Journal of Applied Biological Chemistry
    • /
    • 제62권4호
    • /
    • pp.347-353
    • /
    • 2019
  • Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

청간해주환(淸肝解酒丸)의 알코올 유도 뇌신경세포 손상에 대한 보호 효과 (Effect of Chungganhaeju-hwan in Ethanol-induced Neuronal Cell Damage)

  • 주미선;김효근;조해정;심재종;전용준;오명숙
    • 대한본초학회지
    • /
    • 제26권3호
    • /
    • pp.75-82
    • /
    • 2011
  • Objectives : In this study, we evaluated the effect of Chungganhaeju-hwan(CGHJH) on hydrogen peroxide($H_2O_2$)-induced and ethanol(EtOH)-induced neuronal damage in vitro and in vivo, respectively. Methods:We carried out the anti-oxidant effects of CGHJH against hydrogen peroxide($H_2O_2$)-induced toxicity in HT22 and PC12 cells using thiazolyl blue tetrazolium bromide. Then, to investigate the protective effect on CGHJH against EtOH-induced memory impairment and hippocampal cell damage in male ICR mice, we performed novel object recognition test(NORT), and analysed the brain tissues after immunohistochemistry and western blotting. Results:CGHJH showed protective effect from $H_2O_2$-induced cell toxicity at doses of $1\sim100{\mu}g$/mL in both HT22 and PC12 cells. CGHJH had also recovery effect from EtOH-induced memory impairment in ICR mice from NORT and it protected hippocampal cells against EtOH toxicity in the result of cresyl violet and NeuN immunoreactivity. Conclusion : These results demonstrate that CGHJH has protective effect in neuronal cells against $H_2O_2$ and EtOH toxicities and this effect could be a main role of recovery effect on EtOH-induced memory loss.

산민들레 종자의 발아특성 연구 (Germination Characteristics of Taraxacum ohwianum Kitam.)

  • 신소림;임윤경;권혁준;김수영
    • 한국약용작물학회지
    • /
    • 제24권3호
    • /
    • pp.177-182
    • /
    • 2016
  • Background: Many Taraxacum spp. plants are widely used in medicine, but some of them have propagation problems, such as strong dormancy and poor germination rates. This study investigated the effects of temperature, gibberellic acid ($GA_3$), and potassium nitrate ($KNO_3$) on seed germination in Taraxacum ohwianum. Methods and Results: The seeds (NIBRGR0000135524) were exalbuminous, and their length and width were $4.54{\pm}0.032mm$ and $0.97{\pm}0.029mm$, respectively. Various germination temperatures were tested (15, 20, 25, and $30^{\circ}C$), the optimum temperature for germination was found to be $20^{\circ}C$ (31.3%). High temperature ($30^{\circ}C$) resulted in non-typical seedlings (thickened and crumpled cotyledons, with a restricted root system). To increase the germination capacity, $GA_3$ (200, 500, and $1,000mg/{\ell}$) or $KNO_3$ (20, 50, 100, 200, and $500mg/{\ell}$) solutions were used as pre-soaking solutions instead of water. The $GA_3$ treatments increased the germination percentage and rate, but germination percentage was higher with the $KNO_3$ treatments. Under the $50-200mg/{\ell}\;KNO_3$ treatments, the germination percentage exceeded 80% after 12 days, and $50mg/{\ell}\;KNO_3$ was notably effective (91.2% after 15 days). Conclusions: T. ohwianum seeds showed improved germination at low temperatures. The best method for germination was presoaking in $50mg/{\ell}\;KNO_3$ for 24 h, in the dark at $4^{\circ}C$, and then incubating the germinated seed at $20^{\circ}C$ for 15 days.

구선왕도고가 전지방세포(前脂肪細胞) 3T3-L1의 증식(增殖), 항보체활성(抗補體活性) 및 세포독성(細胞毒性)에 미치는 영향 (Effects of Kuseonwangdogo on the Proliferation of Preadipocyte 3T3-L1 Cells, the Anti-Complementary and the Cytotoxic Effects)

  • 최용후;김호경;고병섭;주영승
    • 대한한의학회지
    • /
    • 제20권3호
    • /
    • pp.105-114
    • /
    • 1999
  • To investigate the anti-complementary and cytotoxic effects of oriental prescription, Kuseonwangdogo, on the proliferation of preadipocyte 3T3- L1 cells, we examined biological effects of Kuseonwangdogo. The results obtained were as follows. 1. After 14 days, the body weight of rats treated with Kuseonwangdogo decreased more than that in the control group (p<0.05). However, the weights of liver, spleen and kidney were unchanged. In serum biochemical test, we examined the level of glucose (GLU) and glutamic pyruvic transaminase (GPT). The levels of GOT and CHOL in serum were decreased remarkably by the administration of Kuseonwangdogo (p<0.05). The haematological examination of the tested group showed significant increment of white blood cells (WBC), hemoglobin concentration (HGB), mean corpuscular hemoglobin (MCH) and monocyte (MO). 2. The effect of Kuseonwangdogo on the proliferation of 3T3-L1 cells was tested by the sulforhodamin B(SRB) assay. The high concentration ($100{\mu}l\;and\;200{\mu}l$) of extracts inhibited the proliferation of 3T3- L1 cells. The p-value was <0.01, respectively. 3. The extract of Kuseonwangdogo showed a potent anti -complementary activity. It was suggested that the active principle may be a kind of polysaccharide molecule. 4. The cytotoxic effects of Kuseonwang dogo and its composing herbs in human liver cells (WRL68) and monkey kidney cells (Vero) were examined by the SRB and 3- (4,5- Dimethylthiazol-2-yl) -2,5 diphenyl-2H- tetrazolium bromide (MTT) assay. Cytotoxic effects were not observed.

  • PDF

L-NAME으로 유발된 학습.기억장애와 뇌허혈 손상에 관한 대조환의 효과 (Effects of Daejo-hwan(Tatsao-wan) on L-NAME Induced Learning and Memory Impairment and on Cerebral Ischemic Damage of the Rats)

  • 김근우;구병수
    • 대한한의학회지
    • /
    • 제21권2호
    • /
    • pp.25-36
    • /
    • 2000
  • Objectives : This study demonstrates the effects of Daejo-hwan on learning and memory impairment induced by L-NAME (75 mg/kg) treatment and on cerebral ischemic damage induced by middle cerebral artery (MCA) occlusion in rats. Methods : Daejo-hwan emulsion (73.3 mg/100 g/l ml) was administered to rats along a timed study schedule. The Moms water maze was used for learning and memory test of the rats. The MCA was occluded by using the intraluminal thread method. The brain slices were stained by 2 % triphenyl tetrazolium chloride (TTC) and 1 % cresyl violet solution. Infarct size, neuron cell number and size in penumbra was measured by using computer image analysis system. Results : 1. The escape latency of the Daejo-hwan treated group decreased significantly with respect to the control group. 2.The memory score of the Daejo-hwan treated group showed increase tendency, And the swimming distance was not different between the normal, the control, and the Daejo-hwan treated group. 3. The infarct size of the Daejo-hwan treated group decreased significantly with respect to the control group. 4. The total infarct volume of the Daejo-hwan treated group showed decrease tendency. And the brain edema index of the Daejo-hwan treated group decreased significantly with respect to the control group. 5. The neuron cell number and cell size in penumbra of the Daejo-hwan treated group increased significantly with respect to the control group. Conclusions : According to the above results, it is supposed that Daejo-hwan is clinically applicable to the vascular dementia.

  • PDF

The Experimental Study on Inhibitory Effects of Wild Ginseng Pharmacopuncture Solution on Melanin Biosynthesis

  • Jo, Na Young
    • Journal of Acupuncture Research
    • /
    • 제35권4호
    • /
    • pp.182-186
    • /
    • 2018
  • Background: The purpose of this study was to investigate the effects of wild ginseng pharmacopuncture on melanin production in B16/F10 murine melanoma cells. Methods: To determine the effect of wild ginseng pharmacopuncture solution on B16/F10 cells, cytotoxicity was evaluated using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide (MTT) method. To observe B16/F10 cell growth, death, and morphological changes, Trypan blue solution was used. The Hosoi method was used to investigate the effect of wild ginseng pharmacopuncture solution on melanin production. The Martinez-Esparza method was used to investigate the effect of wild ginseng pharmacopuncture solution on tyrosinase activity. To determine the pathway involved in the melanogenesis in cells exposed to wild ginseng pharmacopuncture solution, a cell-free tyrosinase was used. Results: Following treatment with $200{\mu}L$ of wild ginseng solution, the cell survival rate was $76.32{\pm}2.45%$ which significantly decreased with higher concentrations (${\mu}L$) of wild ginseng (up to $200{\mu}L$). When $100{\mu}L$ of wild ginseng was used, the cell survival rate was $89.95{\pm}2.07%$. No morphological changes or abnormalities were observed in the B16/F10 murine melanoma cells as observed in the Trypan blue test. Melanin production was significantly reduced to $72.17{\pm}3.74%$ at $100{\mu}L$. Using $100{\mu}L$ of wild ginseng solution, tyrosinase activity was significantly decreased to $80.15{\pm}1.05%$. Wild ginseng pharmacopuncture solution reduced melanin production both directly and indirectly. Conclusion: This study suggests that wild ginseng pharmacopuncture solution may be effective in inhibiting melanin production. Further studies are needed to determine safe and effective clinical applications.

대일 수출용 진공포장 냉장등심의 유통기한 설정 (Establishment of Shelf-life of Vacuum Packaged Pork Loins for exporting to Japan)

  • 이무하
    • 한국축산식품학회지
    • /
    • 제18권2호
    • /
    • pp.115-124
    • /
    • 1998
  • This study was carried out investigate hygienic conditions of domestic packing plant for export and the shelf-life by Japanese standards. vacuum packaged chilled port loins provided by five packers(A, B, C, D and E) were transported to Japan in the same condition, Four packers(A, B, C adn d) produced the samples as hygienic as possible and one packer produced loins in the traditional hygienic condition, In TTC(2,3,5-triphenyl tetrazolium chloride) test all samples showed the negative reaction from the date of the quarantine(14 days after slaughtering) to 50 days The numbers of total plate count(TPC) of A, B, C and D packers that were below 108CFU/g until 50 days were accepted by Japanese Standards but TPC of E packer was over 107CFU/g at 30 days. VBN values increased sig-nificantly(P<0.01) in samples of A,B and D packers in the samples of C packer and E packer at 5% and 0.1% level respectively. The pH values of the vacuum packaged chilled pork for export at 14 days after slaughtering were 5.43∼6.00 In sensory evaluation, A, B, C and D packers which improved hygi-enic condition produced the products with good color and appearance. low drip until 40 days storage at $0^{\circ}C$ But as the storage time increase off-flavor occured at 40days in A and D packer and at 50days in B and C packer. In conclusion according to the above results of microbiological physicochemical and sensory evalution the edible periods of Korean vacuum package chilled porks may be estimated to 40∼50 days for the packers who improved hygienic condition and the shelf lives were calculated as 32∼40 days from edible period by a factor of 0.8(safety coefficient) while that of the pork from E packer which was produced in the traditional hygienic condition was estimated to 30 days after slaughtering at $0^{\circ}C$ Therefore the shelf-life of the pork of E packer would be only 24 days.

  • PDF