Effects of epigallocatechin gallate on CoCl_(2)-induced apoptosis in PC12 cells

Yang, Kyu-Ho,Mo, Hyun-Chul,Choi, Nam-Ki,Kim, Seon-Mi,Kim,Won-Jae 大韓小兒齒科學會 2006 大韓小兒齒科學會誌 Vol.33 No.1

Neuronal apoptotic events, consequently resulting in neuronal cell death, are occurred in hypoxic/ischemic condition. This cell death has been shown to be accompanied with the production of reactive oxygen species (ROS), which can attack cellular components such as nucleic acids, proteins and phospholipid. However, the underlying mechanisms of apoptosis induced in hypoxic/ischemic condition and its treatment methods are unsettled. Cobalt chloride (CoCl_(2)) has been known to mimic hypoxic condition including the production of ROS. Epigallocatechin gallate (EGCG). a green tea polyphenol, has diverse pharmacologial activities in cell growth and death. This study was aimed to investigate the apoptotic mechanism by CoCl_(2) and effects of EGCG on CoCl_(2)-induced apoptosis in PC12 cells. Administration of CoCl_(2) decreased cell survival in dose- and time-dependent manners and induced genomic DNA fragmentation. Treatment with 100 µM EGCG for 30 min before PC12 cells were exposed to 150 µM CoCl_(2), being resulted in the cell viability and DNA fragmentation being rescued. CoCl_(2) caused morphologic changes such as cell swelling and condensed nuclei, whereas EGCG attenuated morphologic changes by CoCl_(2). EGCG suppressed the apoptotic peak and a loss of Δψ_(m) induced by CoCl_(2). CoCl_(2) decreased Bcl-2 expression but Bax expression was not changed in CoCl_(2)-treated cells. EGCG attenuated the Bcl-2 underexpression by CoCl_(2). CoCl_(2) augumented the cytochrome c release from mitochondria into cytoplasm and increased caspase-8, -9 and caspase-3 activity, a marker of the apoptotic executing stage. EGCG ameliorated the incruement in caspase-8, -9 and -3 activity, and cytochrome c release by CoCl_(2). NAC (N-acetyl-cysteine), a scavenger of ROS, attenuated CoCl_(2)-induced apoptosis in consistent with those of EGCG. These results suggest, that CoCl_(2) induces apoptotic cell death through both mitochondria- and death receptor-dependent pathway and EGCG has neuroprotective effects against CoCl_(2)-induced apoptosis in PC12 cells. 신경세포자멸사는 저산소 및 허혈환경에서 일어나며 이러한 세포죽은 reactive oxidant species (ROS) 생성을 동반함이 알려져있다. 그러나, 저산소 및 허혈환경에서 일어나는 세포자멸사의 기전 및 그 치료방법은 아직 정립되어 있지 않다. CoCl_(2)는 ROS를 생성하는 등 저산소환경과 유사한 조건을 초래하는 것으로 알려져 있다. Epigallocatechin gallate (EGCG)는 녹차의 polyphenol 성분으로서 세포성장과 죽음에 다양한 약리학적 효과를 나타냄이 알려져 있다. 본 연구는 PC12 세포에서 CoCl_(2)에 의한 세포자멸사기전을 밝히고 이에 미치는 EGCG의 효과를 조사하는데 목적이 있다. Cell viability는 MTT 측정으로 조사되었고, DNA fragmentation은 DNA laddering으로 조사되었다. Bcl-2와 Bax발현 정도는 RT-PCR로, caspase-3와 -9의 활성은 spectrophotometer, caspase-8의 활성은 flow cytometry에 의해 측정되었다. 미토콘드리아에서 세포질로 분비된 cytochrome c는 western blot으로, 분해된 DNA 양과 미토콘드리아 세포막전위 (Δψ_(m))는 FACScan으로 조사되었다. CoCl_(2)투여로 PC12 세포수는 용량 및 시간 의존형태로 감소하였고, genomic DNA fragmentation이 발생하였다. CoCl_(2)투여로 야기된 cell viability의 감소와 DNA fragmentation은 EGCG 전처치에 의해 억제되었다. CoCl_(2)은 세포용적팽창과 condensed nuclei 같은 형태적 변화를 일으켰으며, apoptotic peak, Δψ_(m)감소 및 cytochrome c 유리를 야기하였다. EGCG는 CoCl_(2)에 의한 세포형태변화, apoptotic peak, Δψ_(m)소실 및 cytochrome c 유리를 억제시켰다. CoCl_(2)는 Bcl-2 발현을 감소시켰지만, Bax 발현에는 영향을 미치지 않았다. EGCG는 CoCl_(2)에 의해 야기된 Bcl-2 발현 감소를 억제시켰다. CoCl_(2)는 caspase-3, -8, 그리고 -9의 활성을 증가시켰으며, EGCG는 CoCl_(2)에 의한 세포자멸사를 억제시켰다. 본 실험결과는 PC12 세포에서 CoCl_(2)가 미토콘드리아 의존 및 death receptor의존 기전으로 세포자멸사를 일으키며, EGCG는 세포자멸사기전을 억세지킴으로 신경보호기능을 가짐을 시사하였다.

Terfenadine과 Ketoconazole 병용투여에 의해 유발된 Torsade de Pointes1예

오석규,국향,임수빈,정진원,박양규,박옥규 대한순환기학회 1998 Korean Circulation Journal Vol.28 No.3

양측 쇄골하동맥 및 양측 신동맥 협착을 동반한 Takayasu 동맥염 1예

임선호,임수빈,오석규,이재홍,정진원,박양규,박옥규,소병준,노병석 대한순환기학회 1997 Korean Circulation Journal Vol.27 No.8

Two Newly Recorded Species of the Genus Ypsolopha Latreille (Lepidoptera: Ypsolophidae) from Korea

나솔문,안능호,변봉규,배양섭,Na, Sol-Moon,Ahn, Neung-Ho,Byun, Bong-Kyu,Bae, Yang-Seop Korean Society of Applied Entomology 2016 한국응용곤충학회지 Vol.55 No.1

Ypsolopha속 (나비목:Ypsolophidae)의 2종, Ypsolopha vittella (Linnaeus, 1758)와 Ypsolopha nigrofasciata (Yang, 1977)를 우리나라에서는 처음으로 보고한다. 이 두 종의 진단형질, 기재, 분포, 기주식물, 성충사진 및 암컷 생식기 사진을 제시하였다. Two species of the genus Ypsolopha Latreille are reported for the first time from Korea: Ypsolopha vittella (Linnaeus) and Y. nigrofasciata Yang. The diagnosis, description, distribution, host plants, adult photographs, and pictures of female genitalia are provided.

McNamara분석법에 의한 한국 성장기아동 정상교합자의 두부 방사선 계측학적 연구

양규호 大韓小兒齒科學會 1993 大韓小兒齒科學會誌 Vol.20 No.2

This study was performed by McNarama analysis anddiagnosis of malocclusion of the growing children. The cephalometricradiograms were taken from the growing children at the 8 to 12 years old that consist of 50 males and 50 females with normal occlusion and acceptable profile and 10 items were statistically analysed. The result of this study were as follows: 1. The cephalometric standard measurements of the Korean growing children by age and sex were established by McNarama analysis. 2. All measurements were great in male dominantly, but the mandibular angle was greater in female indicating more steep mandibular inferior border. 3. Increasing with age, the measurements were increased especially in the Nasion Perpendicular to A,Mn.1 to A-Po,Facial Axis Angle,Mandibular Plane Angle,Effective Mabdibular Length,Effective Maxillary Length, and Lower Ant.Facial Height. 4. Compared with the standard measurements of the growing Caucasian suggested by McNaram,the Mx-Mn length was smaller, the lower anterior teeth was located to more anterior slightly, the mandibular angle was so great that mandibular inferior border was steeper, and the facial axis angle was so small that Gnathion was located posteriorly.

Streptomyces의 mutanase 유도에 관한 연구

양규호,정진 大韓小兒齒科學會 1996 大韓小兒齒科學會誌 Vol.23 No.3

The mutan containing α-1,3 bond is an insoluble portion of glucan which is the main component of dental plaque. The secretion of mutanase was assessed with mutan-digesting Streptomyces isolated from soil, and the factors affecting its activity was studied, obtaining the following result. Mutan-digesting Streptomyces was identified as Streptomyces exfoliatus by its characteristics. The effect of dextranase was identified on the media containing blue dextran. A clear zone was produced by Streptomyces exfoliatus on the media containing blue mutan, so showing the secretion of mutanase. A clear zone was significantly produced on the media overlayed with agar containing blue mutan. A clear zone was produced at 2 days after the inoculation of Streptomyces exfoliatus on the media containing below a concentration of 0.025% glucose, at 3 days on the media containing 0.05% glucose, and at 4 days on the media containing 0.1% glucose. Mutan-digestion wasn't appeared early by adding other carbohydrates. The higher concentration of peptone, the later appearance of clear zone was on the media containing below a concentration of 0.1% peptone. These results indicated that the secretion of mutanase was identified from mutan-digesting Streptomyces on the media containing blue mutan, and a clear zone was appeared lately on the media containing higher amount of glucose.

한강변 응급센터에 내원한 익수환자에 대한 임상적 고찰

양진홍,박규남,최승필,김영민,김세경 대한응급의학회 2001 대한응급의학회지 Vol.12 No.2

Background: There are approximately 2,400 drowning deaths reported every year in Korea, but there are few good reports about submerged patients visiting the emergency room, so this study was designed to investigate the clinical characteristics and outcomes of submerged patients treated at an emergency medical center near the Han river. Methods: A retrospective review of 123 cases of submerged patients admitted to our emergency medical center over a 5 year period from 1996 to 2000 was conducted. Results: Among the 123 submerged patients, there were 66 males and 57 females, and their mean age was 33.18. The common cause of submersions was a suicide attempt, followed by lack of attention, unknown etiology, and water sports such as swimming. Most of the patients were transported to ER by 119 ambulance. In-hospital CPR was performed on 27(22%) cases. At initial ECG rhythm of these cases, asystole was noted if 25 patients; VF and pulseless electrical activity, respectively, were noted in two others. Prehospital CPR had been performed on all in-hospital CPR cases except 2. Prolonged ROSC after succesful CPR was noted in 13 cases, The mean duration of CPR was 26 minutes. Five patients survived after CPR. Their neurologic outcomes were assessed on CPC(cerebral performance category). Three patients were categorized in CPC1, the other two patients were categorized in CPC3 and CPC4. Conclusion: Submerged patient's visits to emergency medical center near the Han river were most frequent in summer for suicide purposes. Drowned patients were more hypothermic than other cardiac arrest cases, and resulted in better outcome even with prolonged arrest time, if they were given appropriate prehospital CPR and aggreissive in-hospital resuscitation.

중탄산나트륨(NaHCO₃) 투여가 1500m 런닝기록과 심박수, 혈중 pH, HCO₃??, 젖산농도에 미치는 영향

양정수,이규성,최용어,김학렬 韓國體育大學校附屬 體育科學硏究所 1992 韓國體育大學校附屬 體育科學硏究所論文集 Vol.11 No.1

본 연구에서는 고등학교에 재학중인 학생 8명을 대상으로 중탄나트륨(NAHCO??)투여에 대한 다음과 같은 연구가설을 검증하는 것이었다. 1)중탄산나트륨(300mg/kg.wt)의 섭취는 염기성증(alkalosis)의 유발을 촉진시키는가? 2)중탄산나트륨(300mg/kg.wt)의 섭취는 1500m 런닝기록에 유의한 효과를 나타내는가? 3)중탄산나트륨(300mg/kg.wt)의 섭취는 1500m 런닝수행시 심박수 및 혈중 pH, HCO????, 젖산농도에 유의한 효고를 나타내는가? 중탄산나트륨 투여 전,후간 심박수 및 1500m 런닝기록은 유의한 효과를 나타내지 못하였으나, 혈중 pH는 중탄산나트륨 투여후 1500m 런닝직후에서 유의한 효과(P<0.01)를 나타내었으며 HCO????농도의 경우에 있어서도 투여 60분후(pre-running, P<0.0001)동일한 효과를 나타내는 것이 증명 되었다. 또한 중탄산나트륨 투여후 혈중 젖산농도는 투여전에 비해 투여후에 더욱 크게 증가된 수준(P<0.01)을 나타내었다. 결론적으로, Ergogenic aids로서 중탄산나트륨(300mg/kg.wt)의 투여는 생체내 알카리성 유발을 촉진시키며 혈중 pH, HCO???? 및 젖산농도에 유의한 생리적 효과를 나타내었으나 1500m 런닝기록에 단축효과는 나타내지 못하였다. 이러한 결과가 시사하는바는 운동수행능력의 향상을 위한 NaHCO?? 섭취는 무산소성 운동능력 및 NaHCO??의 투여량과 밀접한 관련성을 나타낸다는 것이다. The purpose of this study was to examine the hypothesis as follows : Firstly, Does the ingestion of sodium bicarbonate(NaHCO??) result. in induce of alkalosis? Secondly, Does the ingestion of sodium bicarbonate result in improving 1500m running records? Lastly, Does the ingestion of sodium bicarbonate result significant effet during 1500rn running in the variables such as heart rate, venous pH, HCO???? and blood lactate concentration? Record of 1500m running and heart rate before and after the sodium bicarbonate ingestion was not shown a significant effect, but it was proved a significant effect, in case after sodium bicarbonate ingestion of venous pH and HC0?? Also, blood lactate concentration was displayed a significantly increased ievels after sodium bicarbonate treatment, compared with sodium bicarbonate treatment before. Conclusively, Treatment of sodium bicarbonate as ergogenic aids results in increasing a alklosis inducing, and a physiological effect of venous pH, HCO???? and blood lactate concentration, but it was not improved a record of 1500m running. We concluded that NaHCO?? ingestion for improvement, of performance was closely related to factors such as total dosage of NaHCO?? and exercise protocol.

수종 치과용 복합레진의 가수분해

양규호,김정란 대한소아치과학회 2000 大韓小兒齒科學會誌 Vol.27 No.2

심미성 수복재로서 임상에서 널리 사용되는 복합레진은 구강내에서 장기간 유지되도록 하기 위하여 그 물성을 향상시킬 필요가 있다. 복합레진의 단점 중 하나로 낮은 마모 저항성을 들수 있으며, 이와 관련된 하나의 인자로 수복물의 표면하 분해(subsurface degradation)가 고려되고 있다. 본 연구에서는 알카리성 용액(0.1N NaOH)에 현재 임상에서 많이 이용되는 Clearfil APX (CL, Kuraray), Heliomolar (H, Vivadent), Surefil (S, Dentsply), Tetric Ceram (TC, Vivadent), TPH (Dentsply), Z100 (3M)의 6종의 복합레진을 보관하였을 때 각 제품의 분해저항성을 평가하고자 하였다. 각 제품당 3개의 시편을 제작하여 무게측정을 한 후 0.1N NaOH 용액에 저장하여 60℃에서 보관하였다. 2주후 제거하여 HCl로 중화, 세척 후 60℃에서 건조하였다. 무게손실, 분해층 깊이, Si 농도등을 기준으로 분해저항성을 평가하여 다음과 같은 결과를 얻었다. 1.각 제품의 무게손실량은 0.45~3.64% 까지 다양하며 Z100, TC, H, S, CL, TPH 순으로 많았다. 2.각 제품의 표면하 분해층 깊이는 제품마다 다양하며, 10.85~73.38㎛의 범위였고, H, Z100, S, TC, TPH, CL 순으로 깊었다. 3.각 제품으로부터 용출된 Si 양은 Z100군 2060ppm으로 가장 많고, H, TPH, CL, TC, S순으로 많았다. 4.무게손실량과 표면하 분해층 깊이사이에 높은 상관관계를 보였다.(r=0.81, p,0.05) 5.주사전자현미경하 관찰시 NaOH용액에 보관한 후 레진기질과 필러사이의 결합의 파괴를 관찰할 수 있었다. 주요어 : 복합레진, 표면하 분해, 알카리성 용액

포도상구균의 약제 내성 및 β-lactamase의 생산에 관하여

양재윤,박상욱,이용진,조규봉,이연태 단국대학교 신소재기술연구소 1994 신소재 Vol.4 No.-

항균제의 오남용으로 내성균의 출혈빈도가 매우 높아져 임상가에서는 환자치료에 어려움이 적지 않다. 특히 많은 균종중에서 병원감염의 문제가 되고 있고 포도상구균에 대한 내성빈도는 환자치료에 중요하다고 판단된다. 따라서, 1992년 12월부터 1993년 2월 사이에 서울대 병원에 내원(來院)한 환자로부터 총 108주의 포도상구균을 분양 받아 항균제 내성검사를 실시하였고 황색포도상구균의 경우는 β-lactamase 생산유무 확인 및 methicillin에 대한 MIC 농도를 비교하였으며, methicillin에 대하여 각기 다른 MIC농도를 갖는 균종에 대한 substrate 분해능 검사를 실시하여 다음과 같은 결론을 얻었다 (1). 항균제 내성검사를 실시한 108주의 포도상구균은 penicillin에 가장 높은 내성을 나타냈다. 황색포도상 구균의 경우는 97%, 표피포도상 구균의 경우 100%, coagulase 음성포도상구균의 경우 95.7%로 나타났다 (2). vancomycin의 경우는 전 균주가 감수성을 나타냈다 (3). methicillin에 대한 내성율은 41.4%로 나타났다. 이 내성균주들은 10가지 검사 항생제중 최고 9가지에 대한 항균제 내성을 나타내어 다약제 내성임을 알 수 있었다 (4). β-lactamase 생산 실험에서는 58균주의 황색포도상 구균중 53주가 양성을 나타내어 91.4%의 결과를 얻었다. β-lactamase 생산균주와 methicillin에 대한 내성균주가 반드시 일치하지는 않았다 (5). Spectrophotometric assay에서는 내성이 높은 균주와 낮은 균주간에 β-lactamase에 의한 substrate 분해능이 서로 유사하게 나와 황색포도상 구균의 methicililin에 대한 내성 획득은 β-lactamase 뿐만이 아닌 penicillin binding protein(PBP)의 구조적 변화로 인한 내성획득도 시사함을 알 수 있다. One hundred and eight Staphylococcus aureus strains were isolated from patients of Seoul National University Hospital from December 1992 to Feburary 1993. All strains were tested for their sensitivities against antibiotics. β-lactamse producing test was also performed in order to know the relationship between the methicillin resistance and β-lactamase production. Its activity was determined by direct spectrophotometric method. Initial velocities of hydrolysis of the antibiotics were monitored at wavelength 323 nm which corresponded to maximal change in absorbance between the unhydrolyzed substrate and the hydrolyzed product. The results are summarized as follows: 1. In antibiotics sensitivity test of 108 isolated strains of Staphylococcus, they showed hightest resistance to penicillin but all of tested strains were susceptible to vancomycin. 2. 41.4% of them showed resistance to methicillin. 3. All methicillin resistant strains were found to be multi-drug resistant, from 3 to 9 antibiotics. 4. 91.4%(53 out of 58) produced β-lactamase, β-lactamase producing strains did not always coincide with the methicillin resistant strains. 5. β-lactamase activities were similar among staphylococus strains, irrespective of their methicillin resistance.