• 한국안광학회지
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• 제8권1호
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• pp.65-71
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• 2003

각막 상피세포는 정상적인 apoptosis과정을 거쳐 세포가 탈락하고 재생한다. 이러한 apoptosis에는 많은 요소들이 관여하여 세포가 사멸하는데 여러 가지 메커니즘이 관여한다. 본 연구에서는 세포고사 인자로 알려진 물질들을 각막 상피세포에서 apoptosis의 유도 여부를 다른 세포와 비교하여 각막 상피세포의 특성을 알아보고자 시행하였다. 본 연구에 이용된 세포고사 유도물질은 recombinant human cytokiness ($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone와 etoposide이다. 이들을 세포에 48시간 처리한 후 세포독성을 MTT assay로 측정하였으며 세포고사는 Hoechst 33342 staining. Annexin V-FITC/PI staining 그리고 DePsipher assay를 이용하였다. 세포고사의 한 경로인 FAS-FAS ligand system에 대한 연구는 immunocytochemistry로 Fas protein 발현 여부를 조사하였다. 모든 유도인자는 농도의존적으로 세포고사를 유도하였는데 Actinomycin D. camptothecin와 etoposide는 제조사의 추천 농도보다 낮은 농도에서 세포고사가 유도되었고 반면에 cytokines, cycloheximide, dexamethasone은 더 높은 농도에서 세포고사를 유도하였다. FAS antigen은 대조군과 처리군 모두에서 발현되었으나 세포고사율에 비례하여 높게 발현되었다. 본 연구 결과 각막 상피세포는 RNA synthesis inhibitor와 topoisomerase inhibitors가 intracellular receptor-activators 보다 세포고사에 민감하게 나타나는 세포의 특성을 보였다.

• Journal of Chest Surgery
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• 제32권2호
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• pp.151-157
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• 1999

배경: 많은 실험적인 연구에서 종양 조직 내의 아포프토시스와 미세 혈관의 생성은 서로 반비례한다고 보고된다. 비소세포 폐암 조직내에서 두 수치의 관계를 조사하여 보았다. 대상 및 방법:조직내의 아포프토시스의 정도는 deoxynucleotidyl trasferase방법으로(Apop Tag In Situ Apoptosis Detection Kit, ONCOR) 측정하였고, 종양내 미세 혈관 밀도는 항 CD 31 항체를 이용하였다. 결과:아포프토시스 지수와 종양내 미세 혈관 밀도 사이에는 통계적으로 유의하게 역 상관관계가 있었다(p = 0.047). 결론: 비소세포 폐암종에서 아포프토시스와 미세 혈관 생성의 정도는 서로 연관이 있다고에 할 수있다. 그리고 종양내의 신생 혈관의 생성이 종양내 아포프토시스의 억제에 기여한다고 유추 할 수 있다.

• 한국패류학회지
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• 제30권3호
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• pp.189-196
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• 2014

A significant production decrease has been witnessed for the Pacific oysters, Crassostrea gigas farmed in the western coastal waters of Korea, presumably by the loss of physiological viability. We evaluated the viability in terms of health indicators, the condition indices and hemocyte apoptosis rates of the oysters inhabiting two representative farming sites, Incheon and Taean each with different environmental variables. In our monthly measurements for the whole year 2013, the indicators were location specific. The condition indices of Incheon were highly variable, 1.67-8.58%, while those of Taean were less, 2.28-5.57%. The condition indices decreased during the spawning seasons, July and September in common. The two oysters exhibited also differed in the apoptotic activities of hemocyte, highly active, 4.03-30.15% for Incheon oysters and less active, 2.87-17.48% for Taean oysters. One thing we could identify was the two measurements were adverse during the critical seasons of spawning, reminiscent of being a useful tool for a health indicator for the oysters. Similar trend was also observed in the time when change in temperature was extreme. The findings in this study are highly indicative of health indicators for the oyster aquaculture.

• Archives of Plastic Surgery
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• 제36권1호
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• pp.11-18
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• 2009

Purpose: Excessive scarring in the forms of keloid and hypertrophic scar could be a consquence of the accumulation of granulation tissue cells due to aberrant control of apoptosis. Verapamil retard extracelluar matrix production and inhibits VEGF production in primary cultured keloid fibroblast. The object of this study was effect of verapamil on VEGF expression and apoptosis in early wound scarring of the rabbit ear. Methods: Full thickness wounds were created on the ventral side of 6 New Zealand rabbits's ear. 16 days after initial wounding verapamil and saline were injected each scars and scars were harvested 1 week, 2 weeks, 4 weeks later. The wounds were stained with hematoxylin and eosin, TUNEL stain, immunohistochemical stain for VEGF and calculated scar elevation index. Results: Histologic analaysis demonstrated significant reduction in inflammation, vascularity and improvement in dermal collagen organization in experimental group. In TUNEL staining apotosis positive cells were increased and immunohistochemial staining of VEGF demonstrated significant reduction of VEGF expression in experimental group. No significant difference was noted in scar elevation index between two groups. Conclusion: This study suggest that intralesional injection of verapamil on early wound scarring of the rabbit ear decreased VEGF production and increased apoptosis and have a benefit on the pathophysiology of scar formation.

• 대한본초학회지
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• 제26권4호
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• pp.149-154
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• 2011

Objectives : Myocardial infarction is caused by heart cell death in a region where coronary arteries supplying blood to the region are occluded. In the present study, we determined whether ethanol extract of Cortex fraxini (HY5053) could attenuate heart injury by inhibiting apoptosis. Methods : Improvement of survival of HepG2 cells, a human hepatocellular carcinoma cell line, and reduction of apoptosis under hypoxic conditions (3% $O_2$) were assessed by trypan blue staining and DNA fragmentation assay, respectively. To assess the impact of HY5053 on the heart injury, Sprague-Dawley rats underwent 1 day of the left anterior descending coronary artery occlusion. HY5053 was given by intraperitoneal injection (200 mg/kg) 1 hr prior to the occlusion. Subsequently, the heart were harvested, excised into 4 slices, and the slices were stained with 2,3,5-triphenyl tetrazolium chloride. Finally, the extent of heart injury represented as ischemic index (%) was assessed. Results : Addition of HY5053 (400 ${\mu}g$/mL) into the culture medium for 1 day under ischemic conditions improved the cell survival by 50%, compared with control (0 ${\mu}g$/mL), consequently delayed apoptosis in 6 hr difference. Also, HY5053 (200 mg/kg) reduced the ischemic index by 44%, compared with control (0 mg/kg). Conclusions : These findings suggested that HY5053 attenuated myocardial infarction by inhibiting apoptosis. Thus, Cortex fraxini could be developed as a novel cardioprotectant to complement a currently available treatment, coronary angioplasty.

• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.1045-1050
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• 2002

The aims of this study were to detect spontaneously occurring apoptosis in cultured porcine ovarian cells, to examine the role of growth hormone (GH), tyrosine kinase (TK), protein kinase G (PKG) and cyclin-dependent kinase (CDK) in the control of this process, and to determine whether the effect of GH on apoptosis is mediated by TK-, PKG- and cdc2-dependent intracellular mechanisms. We studied the action of pGH (10 ng/ml), blockers of TK (genistein, lavendustin, both 100 ng/ml), PKG (Rp-Br-PET-cGMPS, 50 nM; KT5823, 100 ng/ml) and CDK (olomoucine, $1{\mu}g/ml$), as well as combinations of GH with these blockers, on the onset of apoptosis in cultured granulosa cells isolated from antral (3-6 mm) porcine follicles. The functional characteristics of an early apoptotic event, DNA fragmentation, were determined using terminal deoxynucleotidyltransferase (TdT)-mediated dUTP nick end labelling (TUNEL), whilst morphological signs of advanced apoptosis such as pyknosis, chromatin marginalization, shrinkage and fragmentation of nucleus, were detected using routine light microscopy. After culture, some ovarian granulosa cells exhibited DNA fragmentation, which in some cases was associated with morphological apoptosis-related changes (pyknosis, shrinkage and fragmentation of the nucleus). GH significantly reduced the proportion of TUNEL-positive cells. Neither TK nor CDK blockers when given alone, significantly affected the percentage of TUNEL-positive cells although both PKG blockers significantly increased this index. Furthermore, TK and PKG blockers given together with GH, prevented or reversed the inhibitory effect of GH on apoptosis, whilst the CDK blocker olomoucine promoted it. These observations demonstrate apoptosis in porcine ovaries and suggest the involvement of GH, TK, PKG and CDK in the control of this process. They also suggest that the effect of GH on ovarian apoptosis is mediated or regulated by multiple signalling pathways including TK-, PKG- and CDK-dependent intracellular mechanisms.

• Tuberculosis and Respiratory Diseases
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• 제58권5호
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• pp.480-489
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• 2005

연구 배경 : 세포자멸사의 장애는 발암, 암의 진행, 화학치료시 내성 등에 중요한 역할을 한다. XIAP는 IAP군 중에 가장 강력한 caspase 억제제로 알려져 있다. 본 연구는 비소세포폐암에서 XIAP의 면역조직화학적 발현이 종양진행이 환자생존율에 미치는 영향을 확인해 보고자 시행되었다. 방 법 : 수술적 절제가 시행된 80예의 비소세포 폐암종의 조직에서 XIAP의 면역 조직학적 발현을 조사하여 임상병리학적 인자들[환자의 연령, TNM 병기, TNMpT, TNM-pN, VEGF, microvessel density(MVD), PCNA index, apoptotic index (AI)]과 생존율과의 연관을 분석하였다. 결 과 : 편평세포암종 46예 중 42예(91.3%)에서, 샘암종 34예 중 21예(61.8%)에서 양성을 보여 종양의 조직형별 비교시 편평세포암종은 샘암종에 비해 유의하게 높은 XIAP의 발현을 보였다(p=0.001). 각 조직형내에서 비교시 샘암종의 경우 XIAP는 58세이상의 고 연령군 및 VEGF의 발현과 유의한 상관관계를 보였지만(p=0.028, p=0.014, respectively) 편평세포암종의 경우 XIAP는 모든 임상병리학적 인자들과 상관관계를 보이지 않았다. TUNEL 염색으로 결정된 AI는 XIAP 양성군이 $2.5{\pm}4.9%$, XIAP 음성군이 $18.5{\pm}28.9%$로서 후자에서 유의하게 높은 수치를 보였다(p=0.001). AI는 XIAP를 제외한 다른 임상병리학적 인자들과는 상관관계를 보이지 않았다. 생존 여부의 확인이 가능했던 72예에서 XIAP 음성군의 중앙 생존기간은 29.89개월, XIAP 양성군의 중앙 생존기간은 42.5개월로서, 후자에서 술 후 생존 기간은 더 길었지만 통계학적 차이를 보이지는 않았다. 결 론 : 비소세포폐암종에서 XIAP는 종양의 조직형, 특히 편평세포암종에서 높은 발현과, 종양의 AI와 역의 상관관계를 보였다. 그러나 XIAP의 발현이 임상병리학적 예후인자들 및 생존율과 유의한 관련성을 보이지 않은 것은 생체 조직에서 XIAP의 생물학적 역할은 매우 복잡할 수 있다는 것을 암시하므로 향후 이의 생물학적 역할과 관련 물질들에 대한 연구가 좀 더 진행되어야 할 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9171-9176
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• 2014

Aim: P53, the most commonly mutated tumor suppressor gene in all types of human cancer, is involved in cell cycle arrest and control of apoptosis. Although p53 contains several polymorphic sites, the codon 72 polymorphism is by far more common. There are divergent reports but many studies suggest p53 pro/pro SNP may be associated with susceptibility to developing various cancers in different regions of the world. The present study aimed to find any correlation between H. pylori infection and progression of carcinogenesis, by studying apoptosis and the p53 gene in gastric biopsies from north Indian population. Materials and Methods: A total of 921 biopsies were collected and tested for prevalence of H. pylori by rapid urease test (RUT), imprint cytology and histology. Apoptosis was studied by the TUNEL method. Analysis of p53 gene polymorphism at codon 72 was accomplished by PCR using restriction enzyme BstU1. Observation: Out of 921 samples tested 56.7% (543) were H. pylori positive by the three techniques. The mean apoptotic index (AI) in the normal group was 2.12, while gastritis had the maximum 4.24 followed by gastric ulcer 2.28, gastropathy 2.22 and duodenal ulcer 2.08. Mean AI in cases with gastric cancer (1.72) was less than the normal group. The analysis of p53 72 SNP revealed that p53 (Arg/Arg), (Pro /Arg) variant are higher (40.59% & 33.66%) as compared to p53 pro/pro variant (25.74%) inthe healthy population. Conclusions: The North Indian population harbors Arg or Pro/Arg SNP that is capable of withstanding stress conditions; this may be the reason of low incidence of gastric disease in spite of high infection with H. pylori. There was no significant association with H. pylori infection and AI. However, there is increased apoptosis in gastritis which may occur independent of H. pylori or p53 polymorphism.

• Journal of Nutrition and Health
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• 제35권5호
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• pp.505-511
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• 2002

The study was designed to compare the anti-carcinogenic effect of different level of conjugated linoleic acid (CLA) in 1,2-dimethylhydrazine (DMH)-treated rats by determining biomarkers (apoptosis, cell proliferation, eicosanoids, 1,2-diacylglycerol) and phospholipid fatty acid profile in colonic mucosa. Eighty male Sprague Dawley rats weighing 180-220g were randomly divided into 4 groups depending on the content of CLA, i.e. 0.0% CLA, 0.5% CLA, 1.0% CLA, 1.5% CLA. The experimental diet contained protein 21.6%, carbohydrate 54.6%, and fat 14.5% including CLA mixture at different level by weight. The experimental diet was fed for 14 weeks with the initiation of intramuscular injection of DMH, which was injected twice a week for 6 weeks to give total amount of 180 mg/kg body weight. Regardless of the amount of CLA supplemented to diet, CLA significantly increased the apoptotic index but did not have significant effect on cell proliferation in colonic mucosa. CLA was undetected in colonic mucosal phospholipid of rats fed the 0% CLA diet and increased to 5.9mg/g phospholipid in rats fed the 0.5% diet. The apoptotic index was increased by 251% and the 1,2-DAG content was decreased by 57% in rats fed 0.5% CLA. No further changes in these variables were observed when CLA in the diet was raised to 1.0% or 1.5%. However, dietary CLA decreased mucosal levels of prostaglandin (PG)E$_2$, thromboxane (TX)B$_2$, and arachidonic acid in dose-dependent manner. The present data indicate that dietary CLA can inhibit DMH-induced colon carcinogenesis by mechanism probably involving increased apoptosis.

• The Korean Journal of Physiology and Pharmacology
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• 제25권2호
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• pp.147-157
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• 2021

Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 ㎛ microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.