• Title/Summary/Keyword: TUNEL method

검색결과 89건 처리시간 0.025초

마우스 비장세포에서 방사선유도성 아포토시스에 대한 EGCG 효과의 부족 (Lack of EGCG Effects on Radiation-Induced Apoptosis of Mice Splenocytes)

  • 장성순;이희관
    • Radiation Oncology Journal
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    • 제26권3호
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    • pp.173-180
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    • 2008
  • 목 적: 치사선량의 방사선에 의해 유도되는 아포토시스에 대해 항산화제 또는 산화제로 알려진 EGCG가 어떤 효과를 나타내는지를 마우스 비장에서 연구하였다. 대상 및 방법: C57BL/6 마우스를 실험군에 따라 정상대조군, 방사선조사 단독군, EGCG 전처치군(방사선조사 1시간 전에 100 mg/kg 복강주사)으로 분류하고 7 Gy의 단일선량으로 전신조사 하였다. 각 실험군별 비장 내의 아포토시스는 TUNEL assay를 통해 분석하였고, Bax와 Bcl-2 유전자 발현은 Western blotting 방법으로 측정하였다. 결 과: 비장 내 아포토시스는 주로 white pulp에만 국한되어 발견되었으며, 방사선조사 후 8시간째에 최대의 아포토시스지수를 나타냈다. 8시간째에 방사선조사 단독군과 EGCG 전처치군의 아포토시스지수는 53.9%와 52.1%로 유의한 차이를 보이지 않았다(p=0.328). 또한 두군간에 Bax/Bcl-2 ratio (1.64 vs 0.97, p=0.037)와 대조군에 상대적인 Bcl-2 단백질 발현(0.57 vs 0.82, p=0.037)에서는 유의한 차이를 보였으나, Bax 발현(0.86 vs 0.81, p=0.335)에서는 차이를 보이지 않았다. 결 론: 마우스 비장세포에서 EGCG가 방사선에 의해 유도되는 아포토시스를 유의하게 감소시키지 못하였고, 추가적인 아포토시스의 증가를 유발하지도 않았다.

원지(遠志)가 뇌혈류 저하에 의한 흰쥐 뇌조직의 산화적 손상과 해마신경세포 자연사에 미치는 영향 (Effects of Polygalae Radix on Brain Tissue Oxidative Damage and Neuronal Apoptosis in Hippocampus Induced by Cerebral Hypoperfusion in Rats)

  • 구용모;곽희준;권만재;송민철;이지승;신정원;손낙원
    • 대한본초학회지
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    • 제31권1호
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    • pp.7-15
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    • 2016
  • Objectives : Polygalae Radix (POL) has an ameliorating effect on learning and memory impairment caused by cerebral hypoperfusion. In regard to POL's action mechanism, this study was carried out to investigate the effects of POL on oxidative damage and neuronal apoptosis induced by cerebral hypoperfusion in rats.Methods : The cerebral hypoperfusion was induced by permanent bilateral common carotid artery occlusion (pBCAO) in Sprague-Dawley rats. POL was administered orally once a day (130 mg/kg of water-extract) for 28 days starting at 4 weeks after the pBCAO. Superoxide dismutase (SOD) activities and malondialdehyde (MDA) levels in the brain tissue were measured using ELISA method. Expressions of 4-hydroxynonenal (4HNE) and 8-hydroxy-2'- deoxyguanosine (8-OHdG) were observed using immunohistochemistry. In addition, neuronal apoptosis was evaluated with Cresyl violet staining, TUNEL labeling, and immunohistochemistry against Bax and caspase-3.Results : POL treatment significantly increased SOD activities and significantly reduced MDA levels in the cerebral cortex. The up-regulations of 4HNE and 8-OHdG expression caused by pBCAO in the CA1 of hippocampus were significantly attenuated by POL treatment. POL treatment also restored the reduction of CA1 thickness and CA1 neurons caused by pBCAO and significantly attenuated the apoptotic markers including TUNEL-positive cells, Bax, and caspase-3 expression in the CA1 of hippocampus.Conclusions : The results show that POL attenuated the oxidative damage in brain tissue and neuronal apoptosis in the hippocampus caused by the cerebral hypoperfusion. It suggests that POL can be a beneficial medicinal herb to treat the brain diseases related to cerebral hypoperfusion.

위샘종과 위샘암종에서의 세포자멸사와 세포증식 (Apoptosis and Cell Proliferation in Gastric Adenoma and Adenocarcinoma)

  • 이동수;강상범;이승우;남순우;유영경;한석원
    • Journal of Gastric Cancer
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    • 제6권2호
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    • pp.91-96
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    • 2006
  • 목적: 일반적으로 DNA가 손상된 세포들은 사멸되거나 적절히 손상된 부위를 복구하여 항상성을 유지하거나, 손상된 DNA를 가지고 계속 증식하여 결국 암으로 진행한다. 그러므로 세포자멸사와 세포증식의 균형의 변화는 조직 항상성 및 암 발생의 중요한 조절기전이다. 이에 본 연구자들은 위샘종 또는 위샘암종 조직을 대상으로 세포 사멸 및 세포증식의 정도를 보고자 하였다. 대상 및 방법: 내시경적으로 절제된 위샘종 41예, 외과 수술로 절제된 위샘암종 100예를 대상으로 면역조직화학적 검사를 시행하여 Ki-67 labelling 지수를 구하고, TUNEL 방법을 이용하여 세포자멸사 지수를 구하여 위샘종에서의 이형성 정도에 따른 발현도의 차이 및 위샘암종에서의 조직분류 및 병기에 따른 발현도의 차이를 관찰하였다. 결과: Ki-67 labelling 지수는 위샘종 $51.90{\pm}1.45$, 위샘암종 $55.33{\pm}0.94$로서 위샘암종에서 의의 있게 높았다(P<0.05). 세포자멸사 지수는 위샘종 $53.27{\pm}2.67$, 위샘암종 $42.41{\pm}1.32$로서 위샘종에서 의의 있게 높았다(P<0.05). 위샘종에서 이형성에 따른 Ki-67 labelling 지수 및 세포자멸사 지수는 차이가 없었다. 위샘암종에서 Ki-67 labelling 지수 및 세포자멸사 지수는 Lauren 분류법에 의한 장형과 미만형, 조기 위암과 진행성 위암, 림프절 전이 유무, TNM 분류에 따른 각 군 간의 통계학적 차이는 보이지 않았다. 결론: 샘암종에서의 세포자멸사 지수와 Ki-67 labelling 지수에 대한 연구에서 위샘종은 위샘암종보다 좀 더 정적인 결과를 보이고, 위암발생에서는 세포증식이 중요한 역할을 하나 이 두 지수가 위암의 조직학적 분류 및 병기에 따른 예후와는 관련이 없었다.

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가미향사육군자탕(加味香砂六君子湯)이 생쥐 소장(小腸)에서 방사선조사(放射線照射) 후(後) 보호효과(保護效果)와 Apoptosis에 미치는 영향(影響) (Effects of the Gamihyangsayukgunjatang on Radioprotection and Apoptosis in Small Intestines of Mice)

  • 이태업;김진성;윤상협;류봉하;박동원;류기원
    • 대한한방종양학회지
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    • 제6권1호
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    • pp.1-18
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    • 2000
  • The present study was performed in order to evaluate the effects of the gamihyangsayukgunjatang on radioprotection and apoptosis in small intestines of mice after whole body irradiation. Two hundred forty mice were divided into 40 groups according to the radiation dose and the gamihyangsayukgunjatang treatment. The extracts of the herbal medicines were orally administered to each group differently before and/or after irradiation. The gamihyangsayukgunjatang treated groups were divided into 3 groups. Sample Ⅰ was the group treated with the gamihyangsayukgunjatang for 3 days before the radiation, sample Ⅱ was the group treated with the gamihyangsayukgunjatang for 3 days after the radiation. Sample Ⅲ was the group treated with the gamihyangsayukgunjatang for both 3 days before and after the radiation. To analyze the crypt survival, the micradony survival assay was used according to the Withers and Elind's method. To analyze the apptosis, the TUNEL assay was done. The results obtained are a follows : 1. From the microcolony survival assay, the gamihyangsayukgunjatang treated groups showed the radioprotective effect with a statistical significance(p<0.05), as compared to the control group. Comparing the radioprotective effect among the 3 groups, sample III was statistically more significant than sample I and II (p<0.05). Sample I showed no effect. In accordance with the research mentioned above, it is suggested that the radioprotective effect of the gamihyangsayukgunjatang is more useful for the treatment of the radiation injury rather that the prevention. 2. The results of the TUNEL assay showed that the apoptotic index in the gamihyangsayukgunjatang treated group was slightly decreased with no effectiveness, as compared to the control group. According to the above results, it could be suggested that the gamihyangsayukgujatang has a prominent protective effect in mice intestines against the radiation damage. However, the radioprotective effect does not seem to be related to inhibition of the apoptosis.

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Inhibitory Effects of 3-Bromopyruvate on Human Gastric Cancer Implant Tumors in Nude Mice

  • Xian, Shu-Lin;Cao, Wei;Zhang, Xiao-Dong;Lu, Yun-Fei
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3175-3178
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    • 2014
  • Background: Gastric cancer is a common malignant tumor. Our previous study demonstrated inhibitory effects of 3-bromopyruvate (3-BrPA) on pleural mesothelioma. Moreover, we found that 3-BrPA could inhibit human gastric cancer cell line SGC-7901 proliferation in vitro, but whether similar effects might be exerted in vivo have remained unclear. Aim: To investigate the effect of 3-BrPA to human gastric cancer implant tumors in nude mice. Materials and Methods: Animals were randomly divided into 6 groups: 3-BrPA low, medium and high dose groups, PBS negative control group 1 (PH7.4), control group 2 (PH 6.8-7.8) and positive control group receiving 5-FU. The TUNEL method was used to detect apoptosis, and cell morphology and structural changes of tumor tissue were observed under transmission electron microscopy (TEM). Results: 3-BrPA low, medium, high dose group, and 5-FU group, the tumor volume inhibition rates were 34.5%, 40.2%, 45.1%, 47.3%, tumor volume of experimental group compared with 2 PBS groups (p<0.05), with no significant difference between the high dose and 5-FU groups (p>0.05). TEM showed typical characteristics of apoptosis. TUNEL demonstrated apoptosis indices of 28.7%, 39.7%, 48.7% for the 3-BrPA low, medium, high dose groups, 42.2% for the 5-FU group and 5% and 4.3% for the PBS1 (PH7.4) and PBS2 (PH6.8-7.8) groups. Compared each experimental group with 2 negative control groups, there was significant difference (p<0.05); there was no significant difference between 5-FU group and medium dose group (p>0.05), but there was between the 5-FU and high dose groups (p<0.05). Conclusions: This study indicated that 3-BrPA in vivo has strong inhibitory effects on human gastric cancer implant tumors in nude mice.

경남지방의 도태우에 불임과 관련된 난소의 형태학적 관찰 2. 난소낭종의 발생과 낭종형태에 대하여 (Morphological Observations of Ovaries in Relation to Infertility in Slaughtered Cows in Kyungnam Province 2. Incidences and Morphological Findings of Ovarian Cysts)

  • 곽수동;표병민;양재훈;김철호;서득록;고필옥;강정부
    • 한국임상수의학회지
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    • 제19권2호
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    • pp.153-158
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    • 2002
  • Ovaries from total 192 slaughtered cows(154 Korean native cows and 38 Holstein cows) were collected during the slaughtering process in Kimhae, Changyoung and Yangsan abattoirs in Kyungnam province from January 2001 to January 2002. In order to investigate incidence of the ovarian cysts, anatomical, histological observations were performed and also TUNEL methods and PCNA antibody by immunogistochemical methods for diagnostic accuracy of cysts in a few ovaries were applied. Apoptotic positive cells by TUNEL method appeared not or a few in cystic walls but appeared more number in normal large follicular walls and the proliferative positive cells by PCNA antibody appeared numerous in normal large follicular walls but not or a few in cystic walls. The incident rates of ovarian cysts were 19.5% in Korean native cows and 18.4% in Holstein cows. The incident rates of ovarian cysts in Holstein cows were lower than that of Koran native cows. The incident rates of follicular cysts and luteal cysts in Korean native cows were 11.7% and 7.8% respectively. The incident rates of follicular cysts and luteal cysts in Holstein cows were 10.5% and 7.9%, respectively. Higher incidence proportions of ovarian cysts according to seasons in Korean native cows were ordered as spring (29.8%), autumn (21.4%) winter (14.3%) and summer (6.7%). Rates of cows with single cyst and multiple cysts were 63.3%(19 heads /30 heads) and 36.7%(11 heads/30 heads) in 30 cystic Korean native cows, respectively. Cystic cows with corpus luteums were 50.0%(15 heads) in 30 Korean native cows and 42.9%(3 heads) in 7 dairy cows, respectively. Among 15 cystic Korean native cows with corpus luteums, rates of cows with single corpus luteum were 66.7%(10 heads) and rates of multiple corpus luteum were 33.3%(5 heads ), respectively. The average diameter of cysts and corpus luteums in cystic ovaries were 21.0$\times$17.1 mm and 18.1$\times$13.8 mm in 30 Korean native cows and 20.6$\times$17.7 mm and 19.3 $\times$ 14.9 mm in 7 Holstein cows, respectively. So the average sizes of cysts in cystic ovaries were larger than those of corpus luteums.

시판 중인 구강청결티슈의 세포 독성 관찰 (The Cytotoxic Effect of Oral Wet Wipes on Gingival Cells)

  • 정임희;박지현;이민경;황영선
    • 치위생과학회지
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    • 제18권2호
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    • pp.76-84
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    • 2018
  • 구강위생을 위한 간편성과 편리성 때문에 영유아에서 건강취약자 및 일반인에 이르기까지 구강청결티슈 사용이 증가되고 있다. 또한 기본 성분인 정제수 외에도 구강위생에 도움이 될 목적으로 다양한 기능 성분이 첨가된 구강청결티슈들이 시판되고 있다. 하지만 함유 성분에 대한 제공정보가 부족하고 함유량 기준이 마련되지 못하고 있어 구강 환경이 예민한 영유아 등에게 적용하기 위해서는 이들 제품에 대한 연구자료가 필요할 것으로 보인다. 본 연구에서는 구매도가 높은 시판 구강청결티슈 5종에 대한 구강세포 안전성을 확인함으로써 제품사용 시 유의해야 할 정보를 제공하고자 하였다. 시험 결과 피셔프라이스와 닥터케네디 제품 성분은 구강미생물 S. mutans 및 A. actinomycetemcomitans에 대한 항균작용을 나타냈지만 구강상피세포 및 구강섬유세포에도 독성을 나타냈다. 항균작용이 제한적인 궁중비책, 마이비, 아이수 제품 성분은 구강상피세포 및 구강섬유세포에 대한 독성도 낮았다. 피셔프라이스와 닥터케네디 제품 성분에 의한 구강세포 독성은 G2/M phase에서 세포주기 진행 억제와 세포자멸 유도에 의한 것으로 분석되었다. 따라서 구강 환경이 예민한 영유아 등을 대상으로 한 반복적이고 빈번한 사용은 구강세포 독성의 가능성을 높일 수 있다.

Apoptin Induces Apoptosis in Human Bladder Cancer EJ and BIU-87 Cells

  • Zhan, Hui;Wang, Jian-Song;Wang, Hai-Feng;Zuo, Yi-Gang;Wang, Chun-Hui;Ding, Ming-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권1호
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    • pp.135-138
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    • 2012
  • Objective: To investigate whether apoptin is a apoptosis-inducing protein with a potential for bladder cancer therapy. Methods: We constructed a PCDNA3/Apoptin eukaryotic expression vector, and transfected this vector into bladder cancer cell lines BIU-87 and EJ, then observed the results by RT-PCR, transmission electron microscopy, MTT assay and the flow cytometry (TUNEL method). Results: PCDNA3/Apoptin successfully induced a high level apoptosis in both bladder cancer cell lines, compared with the controls (p<0.05). Conclusions: Apoptin can induce high level apoptosis in human bladder cancer EJ and BIU-87 cells, which suggests a potential for human bladder cancer therapy.

Hypersensitive and Apoptotic Responses of Pepper Fruit Against Xnthomonas axonopodis pv. glycines Infection

  • Chang, Sung-Pae;Kim, Young-Ho
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.72.1-72
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    • 2003
  • Generally, plants defend themselves against pathogens by structural and biochemical reactions. Defense structures act as physical barriers and inhibit the pathogen from gaining entrance and spreading through the plant. Xanthomonas axonopodis pv glycines, the causal pathogen of bacterial pustule of soybean, causes hypersensitive response (HR). When pepper fruits were inoculated with X. axonopodis pv. glycines, in situ, time-series defense-related structural changes occurred in the inoculated sites. Early responses were programmed cell death (PCD), characterized by condensation and vacuolization of the cytoplasm, condensation of nuclear materials, and fragmentation of the nuclear DNA, which were observed by transmission electron microscopy. Nuclear fragmentation was proven by TUNEL method under confocal laser scanning microscopy and DNA laddering through eletrophoresis. At later stages, plant responses were cell elongation and cell division, forming a periderm-like boundary layer that demarcated healthy tissues from the inoculation sites. Using several stains such as toluidine blue, sudan IV, annexin V, and phloroglucinol-HCl, defense-related materials and structural changes were also examined.

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Adenovirus-Mediated Antisense Expression of Telomerase Template RNA Induces Apoptosis in Lung Cancer Cells

  • Song, Joon-Seok;Kim, Sang-Bae;Lee, Young-Ho;Lee, Kyu-Wan;Jung, Hak-Huyn;Kim, Mee-Hye;Kim, Kyung-Tai;Brown, Robert;Kim, Young-Tae
    • Journal of Microbiology and Biotechnology
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    • 제12권1호
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    • pp.89-95
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    • 2002
  • Telomerase is a ribonucleoprotein complex, whose function is to add telomeric repeats $(TTAGGG)_n$ to chromosomal ends and is also known to play an important role in cellular immortalization. Telomerase is highly active in most tumor cells, yet not in normal cells. Therefore, it may have possible applications in cancer gene therapy. Telomerase consists of two essential components; a telomerase RNA template (hTR) and a catalytic subunit (hTERT). The current study attempted to inhibit the "open" part of the human telomerase RNA (hTR) with an antisense sequence-expressing adenovirus. It was found that the antisense telomerase adenovirus suppressed the telomerase activity, tumor cell growth, and survival in vitro. Furthermore, FACS analysis and TUNEL assay suggested that the reduce viability was mediated through the induction of apoptosis, indicating that this approach might be a useful method for suppressing cancer growth in targeted cancer gene therapy.