Prostaglandin A₂-induced Apoptosis is Not Inhibited by Heme Oygenase-1 in U2OS Cells

Kyoung-Won Ko(고경원),Sun-Young Lee(이선영),Ji-Hyun Ahn(안지현),Jaetaek Kim(김재택),In-Kyung Kim(김인경),Ho-Shik Kim(김호식) 한국생명과학회 2008 생명과학회지 Vol.18 No.11

Prostaglandin A₂ (PGA₂)는 사람 골육종 세포인 U2OS 세포주에서 apoptosis와 heme oxygenase (HO)-1의 발현을 함께 유도하였다. PGA₂에 의한 apoptosis는 HO-1의 과도한 발현이나 HO-1에 대한 small interfering RNA에 의한 발현저하에 의하여 변동되지 않았으나 H₂O₂에 의한 세포사망은 HO-1의 발현 수준에 반비례하여 변동되었다. 또한 thiol antioxidant인 N-acetyl-L-cysteine (NAC)은 PGA₂에 의한 세포사망과 HO-1의 발현 증가를 모두 차단하였지만, non-thiol antioxidant인 butylated hydroxyanisole (BHA)과 ascorbic acid는 세포사망과 HO-1의 발현 유도를 차단하지 않았다. 이와 같은 결과들은 PGA₂는 산화성 손상에 의해서가 아니라 PGA₂의 thiol-reactivity에 의하여 apoptosis와 HO-1의 발현을 유도하며, HO-1의 발현은 PGA₂에 의한 apoptosis와는 독립적인 현상이거나 기능적으로 apoptosis 유도의 하부에 위치하고 apoptosis의 진행에는 기여하지 않을 것이라는 것을 시사해 준다. Prostaglandin A₂ (PGA₂), one of cyclopentenone PGs, induced both apoptosis and heme oxygenase (HO)-1 expression in U2OS cells. PGA₂-induced apoptosis was not perturbed by either over-expression or knock-down of HO-1, whereas H₂O₂-induced cell death was inversely modulated by the expression level of HO-1. In addition, N-acetyl-L-cysteine (NAC), a thiol antioxidant, blocked both apoptosis and HO-1 expression induced by PGA₂. But, non-thiol antioxidants like butylated hydorxyanisole (BHA) and ascorbic acid did not block either apoptosis or HO-1-induction. Taken together, these results suggest that PGA₂ induces both apoptosis and HO-1 expression, which are critically related to the thiol-reactivity of PGA₂, but not oxidative stress, and HO-1 expression may be independent or functionally located downstream of apoptosis by PGA₂ without contribution to apoptosis progression.

Nutlin-3 induces HO-1 expression by activating JNK in a transcription-independent manner of p53

CHOE, YUN-JEONG,LEE, SUN-YOUNG,KO, KYUNG WON,SHIN, SEOK JOON,KIM, HO-SHIK Spandidos Publications 2014 International journal of oncology Vol.44 No.3

A recent study reported that p53 can induce HO-1 by directly binding to the putative p53 responsive element in the HO-1 promoter. In this study, we report that nutlin-3, a small molecule antagonist of HDM2, induces the transcription of HO-1 in a transcription-independent manner of p53. Nutlin-3 induced HO-1 expression at the level of transcription in human cancer cells such as U2OS and RKO cells. This induction of HO-1 did not occur in SAOS cells in which p53 was mutated and was prevented by knocking down the p53 protein using p53 siRNA transfection, but not by PFT-alpha, an inhibitor of the transcriptional activity of p53. Accompanying HO-1 expression, nutlin-3 stimulated the accumulation of ROS and the phosphorylation of MAPKs such as JNK, p38 MAPK and ERK1/2. Nutlin-3-induced HO-1 expression was suppressed by TEMPO, a ROS scavenger, and chemical inhibitors of JNK and p38 MAPK but not ERK1/2. In addition, nutlin-3-induced phosphorylation of JNK but not p38 MAPK was inhibited by TEMPO. Notably, the levels of nutlin-3-induced ROS were correlated with the mitochondrial translocation of p53 and this induction was prevented by PFT-beta, an inhibitor of the mitochondrial translocation of p53. Consistent with the effect of the ROS scavenger and MAPK inhibitors, PFT-beta reduced HO-1 expression and the phosphorylation of JNK induced by nutlin-3. In the experiments of analyzing cell death, the knockdown of HO-1 augmented nutlin-3-induced apoptosis. Collectively, these results suggest that nutlin-3 induces HO-1 expression via the activation of both JNK which is dependent on ROS generated by p53 translocated to the mitochondria and p38 MAPK which appears to be stimulated by a ROS-independent mechanism, and this HO-1 induction may inhibit nutlin-3-induced apoptosis, constituting a negative feedback loop of p53-induced apoptosis.

항암제 AG60이 흰생쥐 위점막 상피세포의 DNA합성에 미치는 영향 : 자기방사법적연구

박경호,안의태,고정식,하상선,한영복,정영신,홍은경,유보림,김상건,이경영,강종구 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.1

부신절제후의 위점막 점액세포의 미세구조

고정식,양남길,구광일,안의태,박경호 순천향대학교 1989 논문집 Vol.12 No.3

The experiment was conducted on the male Sprague-Dawley rats weighing about 250g each, to study the fine structural changes of the gastric mucus-secreting cells following bilateral adrenalectomy. Adrenalectomy was performed by dorsal approach method under ether anesthesia. Animals are fed ad libitum after the operation. The normal and the adrenalectomized rats were sacrificed on the 1st, 3rd, 5th and 7th day after the operation. For the electron microscopic observation, gastric mucosae were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde followed by post fixation in 1% osmium tetroxide. The ultrathin sections were contrasted with uranyl acetate and lead citrate, and examined with JEM 100CX Ⅱ electron microscope. Obtained results were follows: 1. The gastric surface mucous cells of adrenalectomized rat contained mucous granules of less electron density than those of the normal rat contains. And mucous granules of adrenalectomized rats are usually placed apart from the surface plasma membrane. 2. Goblet cells of adrenalectomized rat exhibited well formed microvilli as compared with those of normal rat. 3. Cell differentiation in the isthmic area was generally seem to be delayed following adrenalectomy. 4. Golgi complexes of goblet cells were influenced by adrenalectomy, showing flattening or disorganization of cisterns, non-typical vacuoles, scanty vesicles, etc. 5. Considering the above findings, adrenalectomy probably causes the dysfunction of the gastric mucus cell that results in delayed mucus formation and secretion.

분리된 뇌조직의 수초화세포에 관한 연구

고선용,안의태,양남길,고정식,박경호 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.2

중추신경계통의 신경세포들은 긴 축삭돌기로서 다른 신경세포와 연접을 이루기 때문에 뇌손상시 나타나는 반응도 신경로의 방향과 관계가 깊다. 대부분의 긴 축삭은 수초집으로 둘러싸여 보호를 받고 있고 또 수초는 축삭의 대사활동에 중요한 기능을 발휘한다. 뇌가 손상받아 축삭들이 절단될 때 퇴행되는 축삭을 둘러싼 수초집의 다양한 변화는 손상부위의 괴사과정, 치유과정 및 부분적 재생기능에 중요한 요소가 된다. 본 실험에서는 뇌속에 주위의 다른 신경세포와 의 연결이 단절된 뇌조직의 기둥을 만들고 그속에서 일어나는 형태학적 반응 가운데서 특히 수초구조물의 특수한 반응을 연구하였다. 3Kg 정도의 고양이를 nembutal로 마취시키고 뇌정위고정대에 고정시킨 다음 전두골의 일부를 절개한 후 직경 5mm 크기의 원통형칼날을 내려 눌러서 뇌조직의 기둥이 주변으로 부터 분리되도록 했다. 이때 부위선택은 뇌지도를 이용하여 중간줄무늬체동맥이 아래로 부터 올라오는 것이 손상되지 않도록 해서, 분리된 뇌조직이 계속하여 혈액공급을 받을 수 있도록 하였다. 3~4주동안 생존 시킨 후 1% glutaraldehyde 와 1% paraformaldehyde 용액으로 뇌를 고정시켰고 일반적인 술식으로 표본을 작성하여 전자현미경으로 관찰하였다. 관찰 결과 손상된 줄무늬체기저핵 안에서는 많은 수초구주물들이 불규칙한 모습을 지녔으며, 특히 수초집이 팽되되고 심하게 쭈그러드는 것들이 많았으나 이때도 수초의 기본구조는 흐트러지지 않고 정상에 가까운 것이 많았다. 이는 비록 절단된 축삭이 괴사하여 소멸되더라도, 이를 둘러싼 수초집을 만들고 유지해 온 희돌기아교세포가 건재하기 때문에 아직도 기본구조를 유지한 것으로 생각되었다. 특히 전체적인 모양 심하게 변하는 것은 희돌기아교세포가 오히려 활성화되어 새로운 변형을 시도하는 것으로 생각되며, 그 대표적인 예로서 여유분의 수초집이 신경종말을 비롯한 다른 구조물을 흡인하거나 둘러싸는 현상을 보였다. 가장 특이한 경우로는 거대화된 수초집 속에 상당수의 세포들이 들어 있는 수초화세포의 출현이었다. 이들 세포의 성분을 확정하기는 어려우나 형태학적 특징과 세포의 이동특성, 포식형태등으로 볼때 이 세포들이 별아교세포 성분인 것으로 판단하였다. 이와 같은 별아교세포의 수초화는 이 세포들이 손상받은 뇌조직이 치유과정에서 수초집 안팎의 섬유화를 유도하고, 새로이 재생되려는 축삭의 자라나갈 자리를 차지하므로써 손상된 축삭의 재생을 막는 원인이 될 수 있다고 결론지었다. Since the neurones in the central nervous system make contact with other neurones through their long axons, the tissue reactions following brain damage are complicate, and the reactions appear largely along the course of transected axons. Myelin sheaths around transected axons have to show curious reactions. because they belong to oligodendroglia instead of belong to component. In this experiment, a tissue column of fundus striati nucleus was made by pressing a tube-knife of 5 mm diameter down toward the basal forebrain area, where the middle striatal arteries penetrate the brain. Three to four weeks after the stereotaxic isolation, tissue blocks are punched out from fundus striati nucleus. Ultrathin sections are obtained through the observations are discussed and the following results are obtained. Many axons and their myelin sheaths showed various transformations, including the redundant myelin sheaths and myelinated cells. Many redundant myelin sheaths are wrapped around, or sucked in the axon terminals or other structures. Redundant myelin sheath should be very active structures, even after the degeneration of axon within it, since the myelin sheath is a component of oligodendroglia instead of axonal component. Myelinated cells, the cells dwelled within some giant-swelled myelin sheaths, are observed. They contained relatively light cytoplasm with scanty cell other cell components or axon terminals, within the giant myelin capsule. The structural characteristics and the moldability into the myelin sheath, of these myelinated cells suggested that they are activated astroglial cells. Astroglial activities within or out of the transected myelin may prohibit the new axonal sprouts growing into the myelin sheath during regeneration course of nerve fiber in the central nervous system.

과량의 방사선에 머리부분이 노출된 흰쥐 망막의 속핵층과 신경절세포의 미세구조변화

고정식,최숙경,양남길,안의태,박경호 순천향의학연구소 1995 Journal of Soonchunhyang Medical Science Vol.1 No.2

이 실험은 머리부분이 많은 양이 방사선에 노출되었을 때, 망막 속핵층과 신경세포의 미세구조적 반응을 관찰하기 위하여 시행하였다. 실험동물로는 체중200-250g의 흰쥐를 사용하였으며, 마취상태에서 머리부분에만 방사선을 조사였다. 방사선조사는 선형가속기(Mitsubishi Linear Accelerator, ML-4MV)를 이용하였으며 1분당 200rad씩 연속 조사하여 3,000rad 또는 6,000rad를 조사한 후 6시간, 2일, 6일에 희생시켰다. 각 동물은 1% glutaradehyde- 1% paraformaldehyde액으로 관류고정한 다음, 안구를 적출하여 톱니둘레를 따라 절단한 후 망막의 일정부위에서 조직을 떼어내었다. 떼어낸 조직은 2.5% glutaradehyde- 1.5% paraformaldehyde 액에 일차고정한 다음 1% osmium tetroxide액에 이차 고정하였다. 고정된 조직은 수세와 탈수과정을 거쳐 araldite혼합액에 포매한 다음 얇은 절편을 작성하여 uranyl acetate와 lead citrate로 염색한 후 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 무축삭신경원은 세포질의 전자밀도가 높은 어두운 세포(type 3)와 전자밀도가 낮은 밝은 세포로 구분되었으며, 밝은 세포는 핵의 윤곽이 매끄럽고 둥근 세포(type 1)와 핵막의 함입현상이 뚜렷한 세포(type 2)로 구분되었다. 2. 3,000rad의 X-선을 조사한 후 6시간군의 경우, 속핵층을 구성하는 세포들과 신경절세포에서는 속핵층을 구성하는 세포들과 신경절세포에서는 사립체와 세포막이 파괴되는 모습이 관찰되었는데 2일군의 경우 그 경향이 가장 심했다. 3. 3,000rad의 X-선을 조사한 후 6일군의 경우, 부챗살아교세포의 영역이 넓어져서 전체적인 모습이 2일군의 것에 비하여 많이 회복되어 보였다. 4. 6,000rad의 X-선을 조사한 후 6일군의 경우, 속핵층을 구성하는 세포들이 기저복합체 근처에서 관찰되었다. 5. 6,000rad의 X-선을 조사한 후 6시간부터, 부챗살아교세포는 미세사가 많고 전자밀도가 높은 변성된 세포들이 자주 관찰되었다. 이상의 결과를 고찰해 보면 머리부분이 3,000rad의 X-선에 노출되면 망막의 속핵층을 구성하는 세포들과 신경절세포는 처음에는 새포질소기관들과 세포막이 손상을 받으나, 6일 후에는 부챗살아교세포의 영역이 넓어져서 전체적인 구조가 많이 회복된다. 그러나 6,000rad정도의 방사선에 노출되면 부챗살아교세포도 변성되어 속핵층과 신경절세포층의 구조가 변성된다고 생각된다. This experiment was performed to study the morphological responses of the cell of the inner nuclear layer and the ganglion cell of the rat retina following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000rads or 6,000rads of radiation in a single dose, respectively. The source was a Mitsubishi linear accelerator ML-4MV. The target to skin distance was 80cm, and the dose rate was 200 rads/min. The experimental groups were sacrificed on 6hour, 2 and 6 days after X-ray irradiations. Under anesthesia, 1% glutaraldehyde-1% paraformaldehye solution was perfused through the heart. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, and in 1% osmium tetroxide, and embedded in araldite mixture. Ultrathin sections were stained with uranyl acetate and lead citrate solutions and were observed with JEM 100CX-II electron microscope. The results were as follow: 1. Three type of amacrine cells were found, according to their electron densities and nuclear shape, i,e,dark amacrine cell with invaginated nucleus(type 3), light amacrine cell with round nucleus(type 1) and light amacrine cell wit invaginated nucleus(type 2). 2. Six hours after exposure to 3,000 rads of X-ray, morphological changes were observed in cytoplasm of the cells of the inner nuclear layer and the ganglion cell: i.e., edematous changes, fragmentation of mitochondria, breakdown of the plasma membrane, etc. 3. Six days after exposure to 3,000rads of X-ray, many of the malformations were seen to be repaired, except the territory of activated Muller cell. 4. Six days after exposure to 6.000 rads of X-ray, cells of the inner nuclear layer were observed near the Bruch's membrane. 5. Six days after exposure to 6.000 rads of X-ray, darkly degenerated Muller cells were frequently observed. From the above results it wluded that 3,000rads of head-irradiation induces damages to the cell organelles in the cells of the inner nuclear layer and the ganglion cell during the first few days, but many of the malformations can be repaired within six days. And 6,000rads of head irradiation leads to renaturation of the inner nuclear layer and the ganglion cell layer of the retina, since the most of the Muller cells are degenerated within a few days following irradiation.

패밀리레스토랑 종업원의 위생과 청결에 관한 중요도 인식

고호석,김선경,김동기,김범진 한국조리과학회 2005 한국식품조리과학회지 Vol.21 No.2

The purpose of this study was to identify factors associated with sanitation and cleanliness regarding the importance of family restaurant employees in the Ulsan and Basin areas. Accordingly, this study conducted a survey questionnaire consisting of 28 measures of food-service as well as major subject descriptors. From the results, the frequency, descriptive, factor, correspondence analysis, major findings and marketing implications could be summarized as follows: First, 6 factors were extracted: cooking and control factor, food and tableware handling factor, personal cleanliness condition factor, purchase and receiving factor, unit food material store factor and Kitchen utensil sanitation factor. KMO and Bartlett'stest statistics showed that the data fitted the factor analysis well. Results of factor analysis, average variance extracted estimates and shared variance showed that the convergent and discriminant validity of 6 factors are supported and Cronhach's alpha showed that the internal consistency of the 6 factors was supported. Second, correspondence analysis indicated that statistically significant relationships existed between some of the family restaurant employee sanitation and cleanliness factors and the family restaurant enterprise.

WIND ENERGY POTENTIAL ASSESMENT and SITING a WIND FARM in CHEJU ISLAND

고경남,김건훈,조경호,고장권,허종철 濟州大學校 産業技術硏究所 1997 산업기술연구소논문집 Vol.8 No.2

The possibility in the practical use and supply of the wind turbine in Cheju island was investigated. The wind data measured in three regions which have a plenty wind energy potential were utilized to quantify the wind power resources by statistical analysis. Power output was calculated for 600 kW wind turbine. The coastal area of Guja was superior to Daejung and Hoichun in wind power energy densities. The calculated capacity factor of 600 kW wind turbine in this site was high as much as 30%. Hoichun site. the intermediate region between the Halla mountain and the sea. was rich in wind power energy resources in winter. Hourly ideal energy density at three regions increased at sunrise, reached the maximum a t 13-16 O'clock, decreased steadily, and finally remained constant at sunset.

Mitomycin C가 생쥐 콩팥소체의 미세구조에 미치는 영향

고정식,안의태,박경호,박대균 순천향대학교 교수학습개발센터 2004 Journal of Soonchunhyang Medical Science Vol.10 No.3

The experiment was performed to study the morphological responses of the renal glomeruli of the mice after administration of mitomycin C. Healthy adult mice weighing about 25gm each were divided into normal and experimental groups. Mitomycin-C(400㎍/kg) was injected subcutaneously to the animals every other day, and animals were sacrificed at 4 days or 6 days following the first injections. Pieces of tissues were taken from renal corticies, prefixed with 2.5% glutaraldegyde- 1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.), and followed by post-fixation with 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH 7.3). Fixed blocks were dehydrated with alcohol and acetone, embedded in araldite mixture, cut with LKB V-ultrotome. The ultratihin sections stained with urany1 acetate and lead cirtrate were observed with a JEM 100CX-Ⅱ electron microscope. In the fourth and seventh day following the first injection of mitomycin C, components of the renal glomeruli of the mice are looked compact since they were filled with the widened the mesangium, and showed narrowing lumen of glomerular capillaries and of urinary spaces. The electron densities of the lamina rara interna and the lamina rara externa were similar to the density of the lamina densa and the expanded lamina rara interna were often seen. and the cisternae of the rough endoplasmic reticulum of the podocytes were slightly swollen, whereas flattened cisternae of the Golgi complexes were frequently observed. From the above results, it was suggested that although small amount of mitomycin C(400ug/Kg) may give damages on the fine structure of the renal glomeruli of the mice.

5-Fluorouracil이 생쥐 콩팥소체의 미세구조에 미치는 영향

고정식,안의태,박경호,박대균 순천향대학교 교수학습개발센터 2004 Journal of Soonchunhyang Medical Science Vol.10 No.3

The experiment was performed to study the morphological responses of the renal glomeruli of the mice after administration of 5-fluorouracil. Healthy adult mice weighing about 25gm each were divided into normal and 5-fluorouracil treated groups. 5-fluorouracil(60mg/kg) were injected subcutaneously to the animals every other day, and animals were sacrificed at 4 days or 7days following the first injections. Pieces of tissues were taken from renal coriticies, prefixed with 2.5% glutaraldehyde- 1.5% paraformaldehyde(0.1M Millonig's phosphate buffter, pH 7.3), and followed by post-fixation with 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH 7.3). Fixed blocks were dehydrated with alcohol and acetone, embedded in araldite mixture, cut with LKB V-ultrotome. The ultrathin sections stained with uranyl acetate and lead citrate were observed with a JEM 100CX-Ⅱ electron microscope. In the fourth day following the first injection of 5-fluorouracil, components of the renal glomeruli of the mice are looked compact since they were filled with the widened the mesangium, and showed narrowing lumen of glomerular capillaries and of urinary spaces. The cisternae of the rough endoplasmic reticulum of the podocytes were slightly swollen, whereas flattened cistrernae of the Golgi complexes were frequently observed. And in the fourth day after the first injection of 5-fluorouracil, the compact arragement of renal golmeruli of the mice were stil observed, but morphological changes of glomeruli were generally recovered in the seventh day. From the above results, it was concluded that the large amout of 5-fluorouracil(60mg/kg)induces slightly cytotoxic effects of the renal glomeruli, but fine structural changes occurred inthe renal glomeruli after injection of 5-fluorouracil were recovered to normal apperance within 1 week.