• 제목/요약/키워드: Triphenyltetrazolium chloride (TTC)

검색결과 27건 처리시간 0.026초

The Role of Aquaporin-4 in Cerebral Edema Formation after Focal Cerebral Ischemia in Rats

  • Song, Young-Jin;Bae, Hae-Rahn;Ha, Se-Un;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • 제41권1호
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    • pp.30-38
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    • 2007
  • Objective : To elucidate the role of aquaporin-4[AQP4] in cerebral edema formation, we studied the expression and subcellular localization of AQP4 in astrocytes after focal cerebral ischemia. Methods : Cerebral ischemia were induced by permanent middle cerebral artery[MCA] occlusion in rats and estimated by the discoloration after triphenyltetrazolium chloride[TTC] immersion. Change of AQP4 expression were evaluated using western blot. Localization of AQP4 was assessed by confocal microscopy and its interaction with ${\alpha}-syntrophin$ was analyzed by immunoprecipitation. Results : After right MCA occlusion, the size of infarct and number of apoptotic cells increased with time. The ratio of GluR1/GluR2 expression also increased during ischemia. The polarized localization of AQP4 in the endfeet of astrocytes contacting with ventricles, vessels and pia mater was changed into the diffuse distribution in cytoplasm. The interactions of AQP4 and Kir with ${\alpha}-syntrophin$, an adaptor of dystrophin complex, were disrupted by cerebral ischemia. Conclusion : The deranged spatial buffering function of astrocytes due to mislocalized AQP4/Kir4.1 channel as well as increased assembly of $Ca^{2+}$ permeable AMPA receptors might contribute to the development of edema formation and the excitotoxic neuronal cell death during ischemia.

사물탕(四物湯)이 Glutamate에 의한 해마세포의 손상에 미치는 영향 (Effects of Samultang on Glutamate-Induced Apoptosis of Hippocampus Cells)

  • 정대영;최철원;문병순
    • 대한한의학회지
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    • 제30권1호
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    • pp.64-75
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    • 2009
  • Objective: This study was designed to investigate the effect of Samultang (SMT) under hippocampus cells ischemia both in vitro and in vivo. Methods: In the in vitro study, HT22 cells, predominantly detected in the cytoplasm, which coincides with the location of the mitochondria, were used as indicators. In the in vivo study, permanent middle cerebral artery occlusion (MCAO) was induced on rats. SMT was given orally 2 h before induction of permanent focal brain ischemic injury. Result: In the in vitro study, SMT had protective effects in glutamate-induced cytotoxicity, which was revealed as apoptosis characterized by chromatic condensation and the loss of mitochondrial membrane potential in HT22 cells. In the in vivo study, TTC (2,3,5-triphenyltetrazolium chloride) staining showed a marked ischemic injury in blood supply territory of the middle cerebral artery (MCA) such as the cerebral cortex and striatum. However, treatment with SMT significantly reduced infarcted volume. SMT increased marked survival of HT22 cells against glutamate-induced cytotoxicity in MTT assay. Conclusion: These results suggest that water extract of SMT provides neuroprotection against ischemic or oxidative injury by inhibition of apoptotic cell death.

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당뇨흰쥐의 국소뇌허혈에 대한 양격산화탕(凉膈散火湯)의 면역조직화학적 연구 (Immunohistochemical Study of Yanggyuksanhwa-tang on Focal Cerebral Ischemia of Diabetic Rats)

  • 부일권;김연섭
    • 동의생리병리학회지
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    • 제21권3호
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    • pp.741-747
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    • 2007
  • This study evaluated neuroprotective effects of Yanggyuksanhwa-tang (YST), which have been known to be efficacy in the treatment of the stroke and diabetes. on focal cerebral ischemia of diabetic rats. On primary experiment, diabetic condition in rats was induced by streptozotocin injection, then, focal cerebral ischemia was induced by the middle cerebral artery occlusion (MCAO) under the diabetic condition. Then neuroprotective effect of YST was observed with changes of infarct size and volume, expressions of c-Fos, Bax, and hypoxia inducible factor (HIF)-1${\alpha}$ in the brain tissues by using 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining and immunohistochemistry. YST treatment showed a significant decrease of infarct size and volume induced by MCAO in diabetic rats. YST treatment showed a significant decrease of c-Fos and Bax positive neurons in cortex penumbra. YST treatment showed a decrease of HIF-l${\alpha}$ positive neurons in cortex penumbra, but it was not significant statistically. These results suggest that YST has effects on neuroprotection against cerebral infarct under diabetic condition. And it is supposed that neuroprotective effect of YST reveals by anti-apoptosis mechanism.

백서의 가역성 및 비가역성 뇌허혈 모형의 실험적 고찰 (Experimental Analysis in the Reversible and Irreversible Cerebral Ischemic Models in the Rat)

  • 송광철;최병연;김성호;배장호;김오룡;조수호
    • Journal of Korean Neurosurgical Society
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    • 제29권7호
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    • pp.853-860
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    • 2000
  • Objective : The purpose of our experimental study was to analysis the advantages and disadvantages in the reversible and irreversible cerebral ischemic models with rats by staining with Neutral Red(NR) solusion, 2% 2,3,5-triphenyltetrazolium chloride(TTC) and Hematoxylin & Eosin(H & E). Methods : We have measured the range of cerebral infarction in the rat to get a suitable ischemic model along the object of study with and without craniectomy. With craniectomy, 9 rats were sacrificed for irreversible cerebral ischemic model by means of ligation at proximal(group I) and distal(group II), and coagulation at proximal(group III) middle cerebral artery. Also, 6 rats were sacrificed for irreversible(group IV) and reversible(group V) cerebral ischemic model using nylon thread without craniectomy. The sizes of infarction were measured by staining the coronal sections of the brain with NR solusion, TTC and H & E. Results : There are no difference of physiological parameters comparing the each group. Cerebral infarction was not observed in group II, but it's volume was largest in group IV. Disadvantages of craniectomy group(I, II, III) are the long duration of operation and cortical damage by procedure. It's advantage is confirmation of the middle cerebral artery occlusion and cessation of blood flow through the operative microscope. In case of ischemic models using nylon thread (group IV, V), it is hard to identify the interruption or recirculation of blood flow through the middle cerebral artery, but the advantage is the simplicity of operative technique which reduces the operation time and minimizes the cerebral damage due to craniectomy. Therefore, it seems important to set up the reversible and irreversible ischemic models by carefully considering advantages and disadvantages listed above. Conclusion : TTC staining seems to be effective since it reflects the histological damage sufficiently and quickly. It is hoped that researches focused on ischemic penumbra, which became popular recently, will be further carried on with use of NR staining, optical microscope and electron microscope.

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식물체(植物體) 조직배양(組織培養)과 제초제(除草劑) 저항성(抵抗性) (Herbicide Resistance in Plant Tissue Culture)

  • 김길웅
    • 한국잡초학회지
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    • 제5권1호
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    • pp.9-13
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    • 1985
  • 식물체(植物體) 조직배양(組織培養)을 통(通)한 제초제(除草劑)에 대(對)한 저항성(抵抗性) 식물체(植物體) 검정(檢定)에 관한 기초적(基礎的)인 자료(資料)를 얻기 위하여 화본과(禾本科) 잡초(雜草)인 피의 배(胚)로부터 카루스 유도(誘導)에 적정(適定)한 생장조절제(生長調節劑) 농도(濃度) 및 적정배지(適定培地)를 선정(選定)하고 이들 유도(誘導)된 카루스에 TTC를 사용하여 succinate dehydrogenase 활성검정(活性檢定)으로 카루스의 생사판별(生死判別) 및 butachlor에 대(對)한 피종류간(種類間)의 반응(反應)으로 변이종(變異種)을 검정(檢定)하는 등 기초자료(基礎資料)를 얻고져 시험(試驗)을 추진(推進)하여 얻어진 결과(結果)는 다음과 같다. 1. 화본과(禾本科) 잡초(雜草)인 피의 배(胚)로부터 카루스를 유도(誘導)하는 데는 MS 배지(培地)가 적정(適定)하였으며 2, 4-D가 IAA보다 생장조절제(生長調節劑)로 효과적(效果的)이였으며 5.5mg/l이 적량(適量)이었고 발아(發芽)가 된 피 종자(種子)로부터는 카루스가 거의 모두 유도(誘導)되었다. 카루스내(內)에 존재(存在)하는 succinate dehydrogenase의 TTC에 대(對)한 활성(活性)은 돌피, 물피 및 강피 모두가 butachlor 처리(處理) 농도(濃度)에 관계(關係)없이 정반응(正反應)을 보여서 유도(誘導)된 카루스가 생존(生存)하고 있음을 확인(確認)할 수 있었다. 3. Butachlor 농도(濃度)에 따른 피 종류별(種類別) 카루스의 중량증가(重量增加)는 $10^{-3}M$의 고농도(高農度)에서는 피 종류(種類)에 관계(關係)없이 고사(枯死)되었으나 저농도(低濃度)인 $10^{-6}M$에서는 강피가 24.6% 억제(抑制)를 보인데 비(比)하여 돌피는 42%의 억제(抑制)를 보여 돌피가 보다 감수성(感受性)을 나타냈다.

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간장발효에 관여하는 효모에 관한 연구 (제2보) -간장 덧 중에 생육하는 효모에 대하여- (Studies on the Yeasts for the Brewing of Soy Sauce(2) -Isolation, identification and classification of the yeasts in the soy sauce mash-)

  • 이택수;이석건;신보규
    • Applied Biological Chemistry
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    • 제13권2호
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    • pp.171-180
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    • 1970
  • 간장덧중에 생육하는 효모를 경시적으로 계수 및 분리동정 하고 TTC 정색에 의하여 유별한 결과는 다음과 같다. (1) 간장덧 1ml중에 생육하는 일반효모의 수는 담금직후 $127{\times}10^3$, 1개월후 $86{\times}10^3$, 3개월후 $356{\times}10^3$, 6개월후 $1250{\times}10^3$, 2년후 $65{\times}10^3$ 개로 나타났고 내염성효모수는 담금직후 0, 1개월후 $40{\times}10^3$, 3개월후 $81{\times}10^3$, 6개월후 $358{\times}10^3$, 2년후 $23{\times}10^3$개 로서 일반효모와 내염성효모 다 같이 담금초에서 숙성기 까지 상승하였다가 후숙기에서 감소하는 경향을 보였다. (2) 간장덧으로 부터 경시적으로 임의 분리한 50주의 효모를 동정한 결과 Saccharomyces rouxii 10주, Saccharomyces marxianus 1주, Saccharomyces rosei 3주, Saccharomyces fermentati 1주, Saccharomyces mellis 6주, Saccharomyces acidifaciens 1주, Saccharomyces pastori 1주, Pichia polymorpha 3주, Hansenula anomala 2주, Hansenula saturnus 1주, Hansenula suaveolens 2주, Nadosonia fulvescens 5주, Debaryomyces hansenii 8주, Debaryomyces nicotianae 1주, Debaryomyces kloeckeri 1주, Torulopsis sake 2주, Torulopsis holmii 1주, Candida pelliculosa 1주등 7속 18종으로 동정 되었다. (3) 담금기간에 의한 효모의 분포는 담금초에 Saccharomyces rouxii, Saccharomyces marxianus, Saccharomyces rosei, Pichia polymorpha, Hansenula suaveolens, Hansenula saturnus, Debaryomyces hansenii, Torulopsis sake, Candida pelliculosa, Debaryomyces nicotianae, Nadsonia fulvescens 등이 생육하고 숙성기에 Saccharomyces rouxii, Saccharvmyces rosei, Saccharomyces fermentati, Saccharomyces mellis, Saccharomyces pastori, Hansenula anomala, Saccharomyces acidifaciens, Debaryomyces hansenii 등이 생육하고 후숙기에는 Saccharomyces rouxii, Saccharomyces mellis, Nadsonia fulvescens, Debaryomyces hansenii, Debaryomyces kloeckeri, Torulopsis sake, Torulopsis holmii 등이 생육하였다. (4) 담금초기에는 TTC white 효모가 많이 나타났고 발효기 와 후숙기에는 TTC red 효모가 50% 이상을 차지 하였다. (5) 동정한 이들 효모를 TTC 정색에 따라 유별한 결과 Saccharomyces mellis, Saccharomyces pastori는 TIC red로, Saccharomyces acidifaciens는 red pink. Hansenula saturnus, Debaryomyces kloeckeri, Torulopsis holmii는 pink, Saccharomyces marxianus, Nadsonia fulvescens는 white로 각각 나타났고 기타는 전보의 결과와 같았다. Saccharomyces rouxii대부분이 red였으나 red pink로 나타난 것도 있었다, (6) 간장덧중에 생육하는 효모의 종류는 국중에 생육하는 효모의 종류와 거의 비슷하게 나타났으나 국효모는 대부분이 내염성이 없었고 간장덧중의 효모는 발효기간의 경과에 따라 내염성 효모수가 증가 되었다.

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Blood-Brain Barrier Experiments with Clinical Magnetic Resonance Imaging and an Immunohistochemical Study

  • Park, Jun-Woo;Kim, Hak-Jin;Song, Geun-Sung;Han, Hyung-Soo
    • Journal of Korean Neurosurgical Society
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    • 제47권3호
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    • pp.203-209
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    • 2010
  • Objective : The purpose of study was to evaluate the feasibility of brain magnetic resonance (MR) images of the rat obtained using a 1.5T MR machine in several blood-brain barrier (BBB) experiments. Methods : Male Sprague-Dawley rats were used. MR images were obtained using a clinical 1.5T MR machine. A microcatheter was introduced via the femoral artery to the carotid artery. Normal saline (group 1, n = 4), clotted autologous blood (group 2, n = 4), triolein emulsion (group 3, n = 4), and oleic acid emulsion (group 4, n = 4) were infused into the carotid artery through a microcatheter. Conventional and diffusion-weighted images, the apparent coefficient map, perfusion-weighted images, and contrast-enhanced MR images were obtained. Brain tissue was obtained and triphenyltetrazolium chloride (TTC) staining was performed in group 2. Fluorescein isothiocyanate (FITC)-labeled dextran images and endothelial barrier antigen (EBA) studies were performed in group 4. Results : The MR images in group 1 were of good quality. The MR images in group 2 revealed typical findings of acute cerebral infarction. Perfusion defects were noted on the perfusion-weighted images. The MR images in group 3 showed vasogenic edema and contrast enhancement, representing vascular damage. The rats in group 4 had vasogenic edema on the MR images and leakage of dextran on the FITC-labeled dextran image, representing increased vascular permeability. The immune reaction was decreased on the EBA study. Conclusion : Clinical 1.5T MR images using a rat depicted many informative results in the present study. These results can be used in further researches of the BBB using combined clinical MR machines and immunohistochemical examinations.

Apple pectin, a dietary fiber, ameliorates myocardial injury by inhibiting apoptosis in a rat model of ischemia/reperfusion

  • Lim, Sun Ha;Kim, Mi Young;Lee, Jongwon
    • Nutrition Research and Practice
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    • 제8권4호
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    • pp.391-397
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    • 2014
  • BACKGROUND/OBJECTIVE: Myocardial cell death due to occlusion of the coronary arteries leads to myocardial infarction, a subset of coronary heart disease (CHD). Dietary fiber is known to be associated with a reduced risk of CHD, the underlying mechanisms of which were suggested to delay the onset of occlusion by ameliorating risk factors. In this study, we tested a hypothesis that a beneficial role of dietary fiber could arise from protection of myocardial cells against ischemic injury, manifested after occlusion of the arteries. MATERIALS/METHODS: Three days after rats were fed apple pectin (AP) (with 10, 40, 100, and 400 mg/kg/day), myocardial ischemic injury was induced by 30 min-ligation of the left anterior descending coronary artery, followed by 3 hr-reperfusion. The area at risk and infarct area were evaluated using Evans blue dye and 2,3,5-triphenyltetrazolium chloride (TTC) staining, respectively. DNA nicks reflecting the extent of myocardial apoptosis were assessed by TUNEL assay. Levels of cleaved caspase-3, Bcl-2, and Bax were assessed by immunohistochemistry. RESULTS: Supplementation of AP (with 100 and 400 mg/kg/day) resulted in significantly attenuated infarct size (IS) (ratio of infarct area to area at risk) by 21.9 and 22.4%, respectively, in the AP-treated group, compared with that in the control group. This attenuation in IS showed correlation with improvement in biomarkers involved in the apoptotic cascades: reduction of apoptotic cells, inhibition of conversion of procaspase-3 to caspase-3, and increase of Bcl-2/Bax ratio, a determinant of cell fate. CONCLUSIONS: The findings indicate that supplementation of AP results in amelioration of myocardial infarction by inhibition of apoptosis. Thus, the current study suggests that intake of dietary fiber reduces the risk of CHD, not only by blocking steps leading to occlusion, but also by protecting against ischemic injury caused by occlusion of the arteries.

Investigation of a Photothrombosis Inducing System for an Observation of Transient Variations in an in vivo Rat Brain

  • Oh, Sung Suk;Park, Hye Jin;Min, Han Sol;Kim, Sang Dong;Bae, Seung Kuk;Kim, Jun Sik;Ryu, Rae-Hyung;Kim, Jong Chul;Kim, Sang Hyun;Lee, Seong-jun;Kang, Bong Keun;Choi, Jong-ryul;Sohn, Jeong-woo
    • Current Optics and Photonics
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    • 제2권6호
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    • pp.499-507
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    • 2018
  • For the spatiotemporally aligned observation of photothrombosis induction and transient variations of in vivo brain stroke, we developed a novel photothrombosis inducing system compatible to a magnetic resonance imaging (MRI) system using nonmagnetic stereotaxic equipment. From the spatial point of view, the system provides a more reliable level of reproducibility of the photothrombosis in each brain. From the temporal point of view, from T1- and T2-weighted in vivo MR (magnetic resonance) images, the transient variations such as incidence, location, and size of the thrombosis are measured quantitatively. In addition, the final variation is observed in the ex vivo brain by TTC (Triphenyltetrazolium chloride) staining based on histological assay and utilized for the verification of the MR images. From the experimental result of the rat brain, the proposed system shows more reliable characteristics for transient variations of brain strokes.

Supplementation with psyllium seed husk reduces myocardial damage in a rat model of ischemia/reperfusion

  • Lim, Sun Ha;Lee, Jongwon
    • Nutrition Research and Practice
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    • 제13권3호
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    • pp.205-213
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    • 2019
  • BACKGROUND/OBJECTIVES: Myocardial infarction (MI) is caused by extensive myocardial damage attributed to the occlusion of coronary arteries. Our previous study in a rat model of ischemia/reperfusion (I/R) demonstrated that administration of arabinoxylan (AX), comprising arabinose and xylose, protects against myocardial injury. In this study, we undertook to investigate whether psyllium seed husk (PSH), a safe dietary fiber containing a high level of AX (> 50%), also imparts protection against myocardial injury in the same rat model. MATERIALS/METHODS: Rats were fed diets supplemented with PSH (1, 10, or 100 mg/kg/d) for 3 d. The rats were then subjected to 30 min ischemia through ligation of the left anterior descending coronary artery, followed by 3 h reperfusion through release of the ligation. The hearts were harvested and cut into four slices. To assess infarct size (IS), an index representing heart damage, the slices were stained with 2,3,5-triphenyltetrazolium chloride (TTC). To elucidate underlying mechanisms, Western blotting was performed for the slices. RESULTS: Supplementation with 10 or 100 mg/kg/d of PSH significantly reduces the IS. PSH supplementation (100 mg/kg/d) tends to reduce caspase-3 generation and increase BCL-2/BAX ratio. PSH supplementation also upregulates the expression of nuclear factor erythroid 2-related factor 2 (NRF2), and its target genes including antioxidant enzymes such as glutathione S-transferase mu 2 (GSTM2) and superoxide dismutase 2 (SOD2). PSH supplementation upregulates some sirtuins ($NAD^+$-dependent deacetylases) including SIRT5 (a mitochondrial sirtuin) and SIRT6 and SIRT7 (nuclear sirtuins). Finally, PSH supplementation upregulates the expression of protein kinase A (PKA), and increases phosphorylated cAMP response element-binding protein (CREB) (pCREB), a target protein of PKA. CONCLUSIONS: The results from this study indicate that PSH consumption reduces myocardial I/R injury in rats by inhibiting the apoptotic cascades through modulation of gene expression of several genes located upstream of apoptosis. Therefore, we believe that PSH can be developed as a functional food that would be beneficial in the prevention of MI.