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.

한의진단명과 진단요건의 표준화 연구 II (표준화 실례) : 2차년도 연구결과 중간 보고

양기상,최선미,최승훈,안규석,박경모,박종현,김성우,신승호,정우열,전병훈,고현,김정범,신상우,김성훈,김동희,권영규,엄현섭,장혜옥 한국한의학연구원 1996 한국한의학연구원논문집 Vol.2 No.1

The diagnostic requirements were suggested and explained regarding the systems of differentiation of symptoms and signs in the second year study of standardization and unification of the terms and conditions used for diagnosis in oriental medicine. The systems were as follows; - differential diagnosis according to condition of body fluid, differentiation of syndromes according to the state of qi and blood, differential diagnosis according to relative excessiveness or deficiency of yin and yang(氣血陰陽津液辨證) - differentiation of diseases according to pathological changes of the viscera and their interrelation - analyzing and differentiating of febrile diseases in accordance with the theory of the six channels(傷寒辨證) The individual diagnosis pattern was arranged by the diagnostic requirements in the following order : another name(異名), notion of diagnosis pattern, index of differentiation of symptoms and signs(辨證指標), the main point of diagnosis, analysis of diagnosis pattern(證候分析), discrimination of diagnosis pattern(證候鑑別), a way of curing a diseases(治法), prescription(處方), herb in common use(常用藥物), diseases appearing the diagnosis pattern(常見疾病), documents(文獻調査). This study was carried out on the basis of the Chinese documents and references.

F형 종골 금속판의 역학적 특성

양규현,박영희,원정훈,김대야 대한골절학회 2004 대한골절학회지 Vol.17 No.2

치태형성 억제세균과 구강내 세균수와의 관계

양규호,오종석,정현주 大韓小兒齒科學會 1999 大韓小兒齒科學會誌 Vol.26 No.3

소아의 구강으로부터 분리한 비수용성 글루캔 형성 억제세균의 Streptococcus mutans 치태 형성에 대한 억제 정도와 분리 세균속의 타액내 농도가 전체 세균의 농도에 미치는 영향을 본 결과 다음과 같은 결과를 얻었다. 구강으로부터 분리한 비수용성 글루캔 형성 억제세균의 효과를 비커 와이어 검사로 본 결과,교정용 와이어상에 형성된 치태의 무게가 Streptococcus mutans 단독배양시 152mg에서 Enterococcus durans,Lactobacillus acidophilus,Streptococcus oralis와의 혼합배양시 각각 4mg,78mg,72mg으로 감소되였다.Streptococcus mutans의 생균수는 단독배양시 ml당3.6×10×10 개에서 Enterococcus durans,Lactobacillus acidophilus,Streptococcus oralis와의 혼합배양시 각각 ml당 1.4×10 ,5.6×10 ,3.8×10 으로 감소하였다.소아로부터 얻은 타액을 BHI agar에 접종하였을 때의 생균수는 4.8×10 에서 1.3×10 이었으나,이러한 전체 세균의 농도는 타액내 Enterococcus, Lactobacillus, Streptococcus mutans를 억제하는 Streptococcus의 농도와 관련이 없었다. 이상의 결과를 종합하면 소아의 구강으로부터 분리한 비수용성 글루캔 형성 억제 세균의 Streptococcus mutans 치태 형성에 대한 억제는 Streptococcus mutans의 증식을 억제하여 일어났으며, Enterococcus,Lactobacillus,Streptococcus mutans를 억제하는 Streptococcus의 구강내 농도는 구강 즉 타액내 전체 세균의 농도에 영향을 미치지 않았다. The inhibitation degree of the isolated on plaque formation of Streptococcus mutans,and the effect of these bacterial genus on the concentration of total bacteria in saliva were assessed with the following. The effectiveness of the isolated on the inhibitation of plaque formation was assessed culturing Streptococcus mutans in the beaker with orthodotic wires.The mean weight of plaque produced on a wire was 152mg in the culture of Streptococcus mutans only,whereas being reduced to 4mg,78mg,or 72mg in the combined culture of Streptococcus mutans and Enterococcus durans,Lactobacillus acidophilus,or Streptococcus oralis.The colony forming units(CFU) of Streptococcus mutans were 3.6×10 per ml in the culture of Streptococcus mutans,only,wheras 1.4×10 , 5.6×10 ,or 3.8×10 per ml in the culture of Streptococcus mutans and Streptococcus mutans and Enterococcus durans,Lactobacillus acidophilus,or Streptococcus oralis.When saliva from children was 4.8×10 to 1.3×10 per ml of saliva.The concentration of Enterococcus durans,Lactobacillus or Streptococcus inhibiting Streptococcus mutans in saliva was not proportioned to that oh total bacteria replicated on brain heart infusion agar. These results indicate that the isolated bacteria inhibited the replication of Streptococcus mutans,resulting into inhibiting the formation of plaque,but the concentration of Enterococcus,Lactobacillus,or Streptococcus inhibiting Streptococcus mutans,in saliva might not affect the total bacterial concentration of saliva.

Reverse Twin Block을 이용한 III급 부정교합의 치료증례

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

The occlusal inclined plane is the fundamental functional mechanism of the natural dentition. Twin blocks are bite blocks that effectively modify the occlusal inclined plane to induce favorably directed occlusal forces by causing a functional mandibular displacement. Reverse twin blocks may be used to correct sagittal relationships in ClassⅢ malocclusion. The upper bite block is designed to procline upper incisors and ClassⅢ intermaxillary or extraoral traction may be also applied. The patient who was treated with reverse twin block, and following results were observed: 1. Anterior crossbite was corrected by labioversion of maxillary incisors and linguoversion of mandibular incisors. 2. Forward maxillary growth was enhanced, and normal molar relationship was established by mesial drift of maxillary 1st molar and distal drift mandibular 1st molar. 3. Lower anterior facial height was increased in the patient who had horizontal growth pattern, and profile was improved.

신사구체 경변증을 나타내는 FGS/NgaKist mice 간장에서 사이토크롬 P4501A1/2 단백질 발현

정규식,현병화,이철호,박상준,최양규,류시윤,이차수,민병길 대한해부학회 1997 Anatomy & Cell Biology Vol.30 No.2

선천성 치아기형을 동반한 Chromosome 9의 Pericentric Inversion에 관한 증례

양규호,모현철 大韓小兒齒科學會 1990 大韓小兒齒科學會誌 Vol.17 No.2

The authors observed a patient with pericentric inversion of chromosome 9 whose karyotype was 46, XY, inv (9) (p13 q13). The patient, 22-month-old male, was referred to the Department of Pedodontics of Chonnam National University with the chief complaint of severe dental caries of maxillary anterior teeth. The purpose of this case report is to observe physical & dental abnormalities in a child with pericentric inversion of chromosome 9 with a brief review of the literatures. The following results were obtained: 1. Pediatric clinical findings were mental retardation, low level of intelligence, verbal and motor retardation, general hypotonia, flat nasal bridge, oblique palpebral fissures, telecanthus, prominent epicanthal fold, low set/ malformed ears, and congenital heart disease. 2. Pedodontic perioral findings were constantly open mouth, relative marcroglossia, ankyloglossia, narrow palate, thumb sucking, anterior openbite, anterior gingival swelling and redness, fusion of upper/lower left and right primary incisors, congenital missing of upper permanents left and right lateral incisors, and nursing bottle caries, but there was no delayed eruption of primary teeth. 3. In family study, We could consider genetic and familial tendency because of elder brother whose clinical findings were psychomotor retardation, autism and cleft lip etc. But we failed to examine the chromosomes of patients and elder brother due to insistent refusal of the mother.

정상교하 아동의 두개안면부 성장에 관한 종적 연구

양규호,박창헌,손정수,김낙현,최남기,김선미,김기백,신혜성 大韓小兒齒科學會 2009 大韓小兒齒科學會誌 Vol.36 No.3

혼합치열기 정상교합 아동 24명(남:14명 여:10명 초진 시 평균 나이 9±1.3세, 평균 관찰 기간: 13±1.3개월)에 대한 성장량을 측정하여 기능적 교정장치의 순수 치료효과를 평가하는데 도움이 되기 위해 3회(5~8개월 간격) 촬영한 측모 두부 방사선 규격 사진에 대한 분석 결과 다음과 같은 결론을 얻었다. 1. 남아는 상악골은 전하방, 하악골은 전방성장하였고, 여아는 상하악골이 전하방 성장하였다(p<0.05). 2. 상하악골의 남녀간 차이에서 수평적 성장상태는 여아가 컸고(A point 여아: 2.39mm, 남아: 1.26mm, p<0.05), 수직적 성장상태는 유의한 차이가 없었다. 3. 상악 전치의 치축은 두개저에 대해서 순측 경사하였고(p<0.01) 하악 전치의 치축은 큰 변화가 없었다. The purpose of this study was to provide the reference data evaluating the treatment effect of orthopedic appliances. The skeletal and dental growth increments were measured in 24 normal mixed dentition children(boys: 14, girls: 10) by three serial lateral cephalograms: initial mean age: 9±1.3 years, mean observation period: 13±1.3 months, Cephalometric changes were analysed. The results were as follows: 1. In boys, the maxilla showed forward and downward growth pattern and the mandible showed forward growth pattern(p<0.05). In girls, the maxilla and mandible showed forward and downward growth pattern(p<0.05). 2. Horizontal growth of both maxilla and mandible in girls was superior to those in boys(A point: girls: 2.39mm, boys: 1.26mm, with p<0.05), whereas vertical growth of both maxilla and mandible in boys was similar to those in girls. 3. The change in tooth axis showed labioversion of upper incisor(p<0.01) and comparatively stable lower incisor position.

데페록사민 전처치가 토끼 심근경색 크기의 감소에 미치는 효과

양관모,오동렬,박승현,박규남,이원재,김형국,황두영,최승필,채장성 대한응급의학회 1998 대한응급의학회지 Vol.9 No.4

Background: Reperfusion of ischemic myocardium has been postulated to result in a specific oxygen radical mediated tissue injury. Iron may liberate during ischemia and we hypothesized that administration of the iron chelator, deferoxamine during ischemia would result in improved recovery after postischemic reperfusion. Purpose: To test whether iron-catalyzed processes contribute to myocardial necrosis during ischemia and reperfusion, deferoxamine was administered to block iron catalyzed hydroxyl radical formation in rabbits. Methods: Eleven rabbits were divided into two groups : control group (n=5) and deferoxamine pretreatment group (n=6). The left circumflex coronay artery was ligated for 30 minutes and reperfused for 180 minutes. Area at risk (AR) was measured by non-stained area with methylene blue injection into left atrium after left circumflex coronary artery ligation. Infarct size was measured by weighing after triphenyltetrazolium chloride staining. Heart rate was measured using electrocardiographic recording and systemic blood pressure was monitored by pressure transducer connected to the catheter in the left ventricle. Results: 1. There was no significant difference of heart rate and blood pressure in deferoxamine pretreatment group compared with control group. 2. There was significant decrease of serum iron concentration after continuous infusion of deferoxamine compared with serum iron concentration before ligation of coronary artery(P<0.05). 3. There was no significant difference of area at risk between control and deferoxamine pretreatment group. 4. Area at necrosis to area at risk was significantly reduced in deferoxamine pretreatment group compared with control group(P<0.05). The results suggest that deferoxamine infusion prior to coronary artery occlusion has a significant benefit in reducing infarct size in this model.

Nd-YAG 레이저 조사와 불소도포가 법당질의 내산성 변화에 미치는 영향에 관한 실험적 연구

양규호,모현철 大韓小兒齒科學會 1992 大韓小兒齒科學會誌 Vol.19 No.1

The purpose of this study was to examine the combined effects of a pulsed Nd-YAG laser and acidulated phosphate fluoried(APF) on acid resistance of bovine tooth enamel. The materials were 54 freshly extracted bovine permanent teeth with intact smooth enamel surfaces and sectioned sagitally and horizontally for 214 experimental specimens. To determine the most effective energy density of laser in increasing acid resistance of tooth enamel, the author divided 70 specimens into 7 groups including unlased control and 6 experimental groups. Among them, 60 specimens for 6 experimental groups were exposed to a pulsed Nd-YAG laser under conditions raged from 10 to 60J/㎠ with an increment of 10J/㎠. Total 70 specimens for unlased control and 6 experimental groups were etched with 1ml of 0.5N HCIO₄ for 30sec at room temperature and then the amount of dissolved calcium in the acidic solution was measured with an atomic absorption specrophotometer. Scanning electron microscopic(SEM) observations were also made to examine the enamel surfaces of the lased teeth with 14 specimens for unlased control and 6 experimental groups. To survey the combined effects of a pulsed Nd-YAG laser and APF on acid resistance of enamel, the author divided 90 specimens into 9 groups according to treatment method(APF, only, APF after lasing, lasing after APF) and fluoride application time(4min, 60min, 24hr). After determination of the most effective laser-APF combination method on acid resistance by the amount of dissolved calcium in the acidic solution, the rest 40 specimens for SEM were divided into 20 groups according to treatment method(untreated control, APF only, the then, the most effective laser+APF method ) and etching time (non-etching, 15, 30, 60, 90 sec). And then, thd surfaces of each treated specimens were also observed with SEM. The results were as follows: 1. The acid resistance of enamel surface increased significantly compared to untreated control at the laser energy density of 40J/㎠ and 50J/㎠, but more increased at 50J/㎠ without statistical difference between both groups. Laser energy density of 40J/㎠ produced the almost uniform fusion of the lased enamel surface and 50J/㎠ produced the fusion of enamel surface while that of 50J/㎠ produced the fusion of enamel surface and fine cracks of a mosaic pattern. 2. In the groups etched for 902 sec, the surface of lasing(50J/㎠) only group showed its prism structure as in the control, APF only group. But a part of ena,el structure was still resistant to acid in the group treated with APF after lasing. 3. Longer incubation with APF did bring about as increase in acid resistance but there was no significant increase in acid resistance from 4min treated group with the same treatment method except the group treated APF for 24hr after lasing(50J/㎠) 4. The group of APF application after lasing(50J/㎠) caused a remarkable increase in acid resistance of the enamel, while group of lasing after APF application showed a lesser effect similar to either APF only or lasing only group.