• 제목/요약/키워드: Apoptosis assay

검색결과 1,370건 처리시간 0.026초

닭의 발생 단계별 세포내 Telomere의 양적 분포양상과 Telomerase 활성도 분석 (The Amount of Telomeres and Telomerase Activity on Chicken Embryonic Cells During Developmental Stages)

  • 조은정;최철환;손시환
    • Journal of Animal Science and Technology
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    • 제47권2호
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    • pp.187-194
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    • 2005
  • Telomeres locate at the end of chromosomes and consist of a tandem repeat sequence of $(TIAGGG)^{n}$ and associated proteins. Telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. This study was carried out to analyze the amount of telomeres and telomerase activity of chicken cells during embryonic and developmental stages. The whole embryos and prenatal tissues such as brain, heart, liver, kidney and testis at different developmental stages were obtained from Korean Native Chicken. The amount of telomeres on embryonic cells was analyzed by quantitative fluorescence in situ hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) assay. Results indicated that the amounts of telomeric DNA on the most embryonic cells were gradually decreased during ontogenesis. Furthermore, the quantity of telomeres was quite different among embryonic tissues according to developmental origin. The relative amount of telomeres has more in regenerative cells such as embryonic disc and testicular cells than in non-regenerative cells such as liver, brain, heart and kidney cells. Telomerase activity was also highly detectable in most chicken cells at early embryonic stages. After 9 days of incubation, however, the telomerase activitie W

야채수프의 CT26 암세포에 대한 항암 및 면역 증강 효과 (Anti-cancer and Immuno-stimulatory Effects of Vegetable Soup on CT26 Cancer Cells)

  • 임현정;박상민;전형광;류기형;박윤제
    • 한국식생활문화학회지
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    • 제32권5호
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    • pp.453-464
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    • 2017
  • Vegetable soup has been reported to have anti-inflammatory, anti-oxidative and anti-cancer effects. In this study, five kinds of vegetable soup were developed using a new manufacturing process and compositional changes in raw material, and anti-cancer and immuno-stimulatory activities were evaluated. Cytotoxicity tests based on MTT assay revealed that all vegetable soups had strong inhibitory effects against CT26 mouse colon cancer cells, with soups including Solomon's seal being most effective based on comparison of $IC_{50}$ values. Apoptosis in response to vegetable soup was occurred by 3-5 fold on cancer cells compared to normal cells. Mouse splenocytes increased by 266-541% in response to addition of vegetable soup in an in vitro proliferation experiment. In co-culture with splenocytes and CT26 cancer cells, splenocytes increased by more than 280% in every vegetable soup treatment, while cancer cells decreased by about 60% and cytokines such as $IFN- {\gamma}$ and IL-12 were secreted from splenocytes in high levels only in response to vegetable soup including Solomon's seal. In conclusion, all vegetable soups developed in this study had anti-cancer effects, and vegetable soup including Solomon's seal showed the strongest anti-cancer and immuno-stimulatory effects. These results suggest that functionality of vegetable soup could be increased by changes in manufacturing processes and raw materials composition.

인체유방암세포에서 Cyclooxygenase-2 활성 및 Prostaglandin E2 생성에 미치는 어성초 추출물의 영향 (Antiproliferative effect of Houttuynia cordata Thunb was Associated with the Inhibiton of Cyclooxygenase-2 Expression in Human Breast Carcinoma Cells)

  • 정일홍;조인주;박철;최영현;박동일
    • 동의생리병리학회지
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    • 제20권3호
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    • pp.690-696
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    • 2006
  • Houttuynia cordata Thunb, well known as 'E-Sung-Cho' in Korea, is a traditional medicinal plant generally used in Oriental medicine therapy. In the present study, we investigated the effect of water extract of H. cordafa (WEHC) on the growth of human breast carcinoma MCF-7 cells. Exposure of MCF-7 cells to WEHC resulted in growth inhibition and apoptosis in a dose-dependent manner as measured by MTT assay and fluorescent microscope. The anti-proliferative effect of WEHC was associated with a dose-dependent up-regulation of cyclin-dependent kinase inhibitor p21 in a p53-independent fashion. We found WEHC decreased the levels of cyclooxygenase (COX)-2 and inducible nitric oxide synthease (iNOS) expression without significant changes in the expression of COX-1 , which was correlated with a decrease in prostaglandin $E_2{\;}(PGE_2)$ synthesis. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. cordata.

Annexin A5 as a New Potential Biomarker for Cisplatin-Induced Toxicity in Human Kidney Epithelial Cells

  • Kwon, Yeo-Jung;Jung, Jin-Joo;Park, Na-Hee;Ye, Dong-Jin;Kim, Donghak;Moon, Aree;Chun, Young-Jin
    • Biomolecules & Therapeutics
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    • 제21권3호
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    • pp.190-195
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    • 2013
  • Cisplatin is a member of platinum-containing anti-cancer drugs that causes cross-linking of DNA and ultimately cancer cell apoptosis. The therapeutic function of cisplatin on various types of cancers has been widely reported but the side effects have been discovered together and nephrotoxicity has been regarded as major side effect of cisplatin. To select candidates for new sensitive nephrotoxicity biomarker, we performed proteomic analysis using 2-DE/MALDI-TOF-MS followed by cisplatin treatment in human kidney cell line, HK-2 cells, and compared the results to the gene profile from microarray composed of genes changed in expression by cisplatin from formerly reported article. Annexin A5 has been selected to be the most potential candidate and it has been identified using Western blot, RT-PCR and cell viability assay whether annexin A5 is available to be a sensitive nephrotoxic biomarker. Treatment with cisplatin on HK-2 cells caused the increase of annexin A5 expression in protein and mRNA levels. Over-expression of annexin A5 blocked HK-2 cell proliferation, indicating correlation between annexin A5 and renal cell toxicity. Taken together, these results suggest the possibility of annexin A5 as a new biomarker for cisplatin-mediated nephrotoxicity.

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
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    • 제62권4호
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    • pp.347-353
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    • 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.

Effects of a Compound Extract from Agrimonia pilosa Ledeb, Grifola umbellata (pers.) Pilat, and Gambogia on Human Gastric Carcinoma MGC-803 Cells

  • Zhao, Li;Zhang, Xiao-Nan;Gu, Hong-Yan;Wang, Jia;Tao, Lei;Mu, Rong;Guo, Qing-Long
    • Food Science and Biotechnology
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    • 제18권1호
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    • pp.103-107
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    • 2009
  • Three traditional Chinese medicines, Agrimonia pilosa Ledeb, Grifola umbellata (pers.) Pilat, and Gambogia, are combined to form a compound extract, AGC. In this study, the in vitro and in vivo inhibitory effects of AGC on human gastric carcinoma MGC-803 cells were demonstrated, and the molecular mechanisms underlying these effects are investigated. Our results indicate that AGC inhibited MGC-803 cell growth in a dose-dependent manner as measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, with an $IC_{50}$ of about $6.045{\pm}0.69{\mu}g/mL$. In vivo, AGC inhibited growth of human gastric carcinoma in xenograft tumors in nude mice, and the inhibitory rate reached 55.2% at 300 mg/kg. The pro-apoptotic activity of AGC was attributed to its ability to decrease the expression of Bcl-2 and Pro-caspase3 and increase the expression of Bax. These results demonstrate that AGC can effectively induce programmed cell death and may be a promising anti-tumor drug in human gastric carcinoma.

Gluconacetobacter spp. 스타터로 발효한 콤부차의 생리활성 (Biological Activities of Kombucha by Stater Culture Fermentation with Gluconacetobacter spp.)

  • 고혜명;신승식;박성수
    • 한국식품영양과학회지
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    • 제46권7호
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    • pp.896-902
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    • 2017
  • 본 연구는 감귤 콤부차의 산업화를 위한 발효 균주 표준화를 위하여 콤부차에서 분리된 3가지 균주(Gluconacetobacter xylinus, Gluconacetobacter medellinensis, Gluconobacter oxydans)를 이용한 감귤 발효액(CK-MOX)의 기능적 특성을 탐색하고자 하였다. CK-MOX 제조 후 15일간 3일 마다 샘플링을 하였으며, 발효 기간에 따른 pH, 산도, 항산화 능력을 평가하였다. 발효에 따라 pH는 감소하였고, 산도는 증가하였다. DPPH, ABTS 라디칼 소거능, ORAC assay를 통한 항산화 능력 측정 결과 발효에 따라 항산화 능력이 향상하는 것으로 나타났으며, 방광암 세포주(EJ 세포)의 생존 억제 및 이동 억제 효과가 있는 것으로 나타났다. 특히 CK-MOX로 유도된 EJ 세포의 사멸에 MAPK pathway의 중추적인 역할을 하는 것으로 알려진 ERK의 발현이 깊이 관여하는 것으로 나타났다.

계혈등 물추출물의 항산화 및 간보호효과 (Antioxidant Effect and Liver Protection Effect of Spatholobi Caulis Water Extract)

  • 이재준;최홍식;김승모
    • 대한본초학회지
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    • 제26권3호
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    • pp.47-56
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    • 2011
  • Objectives : This study investigated whether the water extract of Spatholobi Caulis (SCE) has the ability to protect hepatocyte against oxidative stress induced by tert-butylhydroperoxide (tBHP) in vitro and $CCl_4$ in vivo. Methods : In vitro, HepG2 cells pre-treated with Spatholobi Caulis water extract (1, 3, 10, $30{\mu}g$/ml) for 12h and further incubated with tBHP ($100{\mu}M$) for the next 12h. Cell viability was assessed by MTT assay. In vivo, rats were orally administrated with the aqueous extract of Spatholobi Caulis (SCE; 50, 100 mg/kg) for 4 days and then, injected with $CCl_4$ 1 mg/kg body weight to induce acute liver damage. Results : Treatment with SCE inhibited cell death induced by tBHP, as evidenced by alterations in the levels of the proteins associated with apoptosis:SCE prevented a decrease in $Bcl_2$, and cleavage of poly(ADP-ribose)polymerase and pro-caspase-3. Moreover, SCE inhibited the ability of tBHP to generate $H_2O_2$ production, thereby restoring GSH content. Moreover, SCE treatments in rats effectively decreased liver injuries induced by a single dose of $CCl_4$, as evidenced by decreases in hepatic degeneration and inflammation as well as plasma alanine aminotransferase and lactate dehydrogenase activities. Consistently, treatments of SCE also protected liver in rats stimulated by $CCl_4$, as indicated by restoration GSH and prevention of MDA in the liver. Conclusions : SCE has the ability 1) to protect hepatocyte against oxidative stress induced by tBHP and 2) to prevent $CCl_4$-inducible acute liver toxicity. Present findings may be informative not only in elucidating the pharmacological mechanism of Spatholobi Caulis, but in determining its potential application for oxidative cellular damage in the liver.

6-Hydroxydopamine 유발 SH-SY5Y 세포주 손상에 대한 resveratrol의 신경보호 효과 (Neuroprotective effects of resveratrol on 6-hydroxydopamine-induced damage of SH-SY5Y cell line)

  • 장건천;김형춘;위명복
    • 대한수의학회지
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    • 제54권1호
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    • pp.1-6
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    • 2014
  • Parkinson's disease is known to exhibit progressive degeneration of the dopaminergic neurons in the substantia nigra via inhibition of glutathione metabolism. It is well known that 6-Hydroxydopamine (6-OHDA) induces Parkinson's disease-like symptoms, while resveratrol (3,5,4'-trihydroxystilbene) has been shown to have anti-inflammatory and antioxidant effects. In the present study, we investigated the neuroprotective effects of resveratrol, a phytoalexin found in grapes and various plants, on 6-OHDA-induced cell damage to the SH-SY5Y human neuroblastoma cell line. Resveratrol (5 and 10 ${\mu}M$) inhibited 6-OHDA (60 ${\mu}M$)-induced cytotoxicity in SH-SY5Y cells and induced a reduction of the number of apoptotic nuclei caused by 6-OHDA treatment. Additionally, the total apoptotic rate of cells treated with both resveratrol (10 ${\mu}M$) and 6-OHDA (60 ${\mu}M$) was less than that of 6-OHDA treated cells. Resveratrol also dose-dependently (1, 5 and 10 ${\mu}M$) scavenged reactive oxygen species (ROS) induced by 6-OHDA in SH-SY5Y cells and prevented depletion of glutathione in response to the 6-OHDA-induced cytotoxicity in the glutathione assay. Overall, these results indicate that resveratrol exerts a neuroprotective effect against 6-OHDA-induced cytotoxicity of SH-SY5Y cells by scavenging ROS and preserving glutathione.

영구치 치수 기질세포를 이용한 연골 분화 및 분화 시기에 따른 형태학적 변화 (Morphological evaluation during in vitro chondrogenesis of dental pulp stromal cells)

  • 정주령;김하나;박열;김민정;오영주;신수정;최윤정;김경호
    • Restorative Dentistry and Endodontics
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    • 제37권1호
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    • pp.34-40
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    • 2012
  • Objectives: The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis. Materials and Methods: Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets. Results: Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction. Conclusions: Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.