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.

A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

Ahn Jae-Il,Jang In-Keun,Lee Doo-Hoon,Seo Young-Kwon,Yoon Hee-Hoon,Shin Youn-Ho,Kim Jae-Chan,Song Kye-Yong,Lee Hee-Gu,Yang Eun-Kyung,Kim Ki-Ho,Park Jung-Keung The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

서울의 Penicillinase Producing Neisseria Gonorrhoeae 발생빈도(1994)

김재홍,안진균,정성재,김영태,김중환,김시영,이석종,이홍렬,서호석,김경훈,권혁진,정우권,고우석,이용석,안필수,오준규,오용섭 대한화학요법학회 1996 대한화학요법학회지 Vol.14 No.1

Background : In recent years gonorrhoea has been panendemic and remains one of the most common Sexually Transmitted Diseases in the world, especially in developing countries. Objective & Method : For the detection of a more effective therapeutic regimen and assessing the prevalence of Penicillinase Producing Neisseria Gonorrhoeae(PPNG), Ive have been trying to study the patients who have visited the VD Clinic of Choong-Ku Public Health Center in Seoul since 1980 by means of the chromogenic cephalosporin method. Results : In 1994, 168 strains of N. gonorrhoeae were isolated, among which 109 (64,9% ) were PPNG. Conclustion : Our results suggests that the prevalence of PPNG in Seoul is still increasing.

The Antioxidant Effect of the Rhaponticum uniflorum Extracts on the Oxidative Stress in H₂O₂ induced Human Keratinocytes

So Hee Ahn,Won Yeoung Choi,Ji Won Jeong,Geon Woo Park,Su Yeon Kang,Eun Jeong Kim,Su Rim Sin,Eun Seo Jang,Gye Won Lee,Young Ho Cho 한국식품영양과학회 2021 한국식품영양과학회 학술대회발표집 Vol.2021 No.10

In this study, To investigate the antioxidant activities of Rhaponticum uniflorum extracts (RUE), we assessed scavenging activities on DPPH, ABTS radicals, and reducing power. The DPPH and ABTS radical scavenging activities (SC50) of RUE were 96.97±1.19 μg/mL and 31.26±0.13 μ g/mL, respectively, and the reducing power significantly increased dose-dependent manner. To study the antioxidant effecters of RUE in H₂O₂ induced human keratinocytes, expression of Nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot and qRT-PCR. RUE induced the expression of HO-1, a critical target enzyme of Nrf2, in a concentration-dependent manner. The expression of HO- 1 mRNA was also induced in H₂O₂-treated HaCaT cells but no significant difference in the level of HO-1 mRNA was observed. Taken together, these results suggest that RUE may be useful as a potential ingredient for anti-aging products.

전통누룩 진균류를 이용한 입국의 제조 및 입국곰팡이의 동정

김재호 ( Jae Ho Kim ),권영희 ( Young Hee Kwon ),이애란 ( Ae Ran Lee ),김혜련 ( Hye Ryun Kim ),안병학 ( Byung Hak Ahn ) 한국균학회 2012 韓國菌學會誌 Vol.40 No.4

다양한 향미를 가진 막걸리의 개발을 위해 전통누룩으로부터 분리한 곰팡이로 입국을 제조한 후 품질특성을 분석하여 입국의 규격에 적합하며 이취가 없고 관능이 우수한 9균주를 입국 제조용 우수균주로 최종 선발하였다. 선발된 균주는 Aspergillus oryzae(C1-5-2-2, C20-7-3, CN1.3.1-4, CN16.19.1-1, N152-1, N220-1), Mycocladus corymbiferus (N162-2), Rhizopus oryzae(N20), Lichtheimia corymbifera (N21)로 동정되었으며, 제조한 입국의 산도는 5.0~6.8, 당화력은 128~241sp이었다. Various koji were prepared by fungi isolated from traditional nuruk and their quality characteristics were investigated. Acidity and saccharification power of their koji were ranged in 5.0~6.8 and 128sp~241sp. Nine fungi which were showed good quality and sensory evaluation were identified by analysis of their nucleotide sequences with PCR-amplified 18S rDNA internal transcribed spacer-1(ITS-1) and ITS-4 genes. Among them, six strains were identified as Aspergillus oryzae and the other strains were identified as Mycocladus corymbiferus, Rhizopus oryzae, Lichtheimia corymbifera.

Protective Effect of Heme Oxygenase-1 on High Glucose-Induced Pancreatic β-Cell Injury

Lee, Eun-Mi,Lee, Young-Eun,Lee, Esder,Ryu, Gyeong Ryul,Ko, Seung-Hyun,Moon, Sung-Dae,Song, Ki-Ho,Ahn, Yu-Bae Korean Diabetes Association 2011 Diabetes and Metabolism Journal Vol.35 No.5

<P><B>Background</B></P><P>Glucose toxicity that is caused by chronic exposure to a high glucose concentration leads to islet dysfunction and induces apoptosis in pancreatic β-cells. Heme oxygenase-1 (HO-1) has been identified as an anti-apoptotic and cytoprotective gene. The purpose of this study is to investigate whether HO-1 up-regulation when using metalloprotophyrin (cobalt protoporphyrin, CoPP) could protect pancreatic β-cells from high glucose-induced apoptosis.</P><P><B>Methods</B></P><P>Reverse transcription-polymerase chain reaction was performed to analyze the CoPP-induced mRNA expression of HO-1. Cell viability of INS-1 cells cultured in the presence of CoPP was examined by acridine orange/propidium iodide staining. The generation of intracellular reactive oxygen species (ROS) was measured using flow cytometry. Glucose stimulated insulin secretion (GSIS) was determined following incubation with CoPP in different glucose concentrations.</P><P><B>Results</B></P><P>CoPP increased HO-1 mRNA expression in both a dose- and time-dependent manner. Overexpression of HO-1 inhibited caspase-3, and the number of dead cells in the presence of CoPP was significantly decreased when exposed to high glucose conditions (HG). CoPP also decreased the generation of intracellular ROS by 50% during 72 hours of culture with HG. However, decreased GSIS was not recovered even in the presence of CoPP.</P><P><B>Conclusion</B></P><P>Our data suggest that CoPP-induced HO-1 up-regulation results in protection from high glucose-induced apoptosis in INS-1 cells; however, glucose stimulated insulin secretion is not restored.</P>

H₂O₂로 산화적 스트레스가 유도된 각질 형성 세포에서 흑삼 추출물의 항노화 효과

안소희(So Hee Ahn),이계원(Gye Won Lee),권득상(Deuk Sang Kwon),신현중(Hyun Joong Shin),조영호(Young Ho Cho) 한국식품영양과학회 2021 한국식품영양과학회지 Vol.50 No.10

본 연구에서는 95°C에서 6번 증숙하여 제조한 흑삼 추출물의 항산화 활성과 피부세포에서 산화적 스트레스로부터 보호 효과를 확인하고, 항산화 효소 발현, 주름개선 및 보습관련 단백질 발현에 미치는 영향을 측정하였다. 흑삼 추출물(BGE-A6)의 항산화 활성을 평가하기 위하여 DPPH, ABTS, superoxide anion 라디칼 소거 활성 및 환원력을 측정하였다. 그 결과 증숙 후 흑삼 추출물이 증숙 전에 비하여 DPPH 라디칼 소거 활성은 4배, ABTS 라디칼 소거 활성은 5배 정도 항산화 활성이 증가하는 것으로 나타났다. 피부 각질 형성 세포에서 산화적 스트레스로부터 BGE-A6의 세포보호 효과를 측정하기 위하여 BGE-A6를 1시간 전처리하고 0.5 mM H₂O₂를 6시간 동안 처리하여 분석한 결과, H₂O₂ 처리로 증가된 ROS 생성이 BGE-A6를 100 μg/mL 농도로 처리한 경우 약 40% 정도 감소되는 것으로 나타났다. 또한, BGE-A6의 ROS 소거 효과를 규명하기 위하여 세포 내 존재하는 항산화 효소의 발현량을 western blot으로 평가한 결과 H₂O₂에 의해 유도된 산화 스트레스로부터 감소된 항산화 효소(HO-1, CAT, SOD)의 발현을 농도 의존적으로 회복시켰다. 또한, BGE-A6의 주름 개선 효과를 확인하기 위하여 콜라겐 발현 및 MMP-1 발현 억제 정도를 ELISA와 western blot으로 측정한 결과 0.5 mM H₂O₂를 처리하였을 때 감소된 콜라겐 생성량이 BGE-A6를 100 μg/mL 농도로 처리할 경우 H₂O₂ 처리에 의해 감소된 콜라겐 생성량을 약 20% 정도 회복시키는 것으로 확인되었다(P<0.05). 0.5 mM H₂O₂를 처리하였을 때 증가된 MMP-1의 생성이 BGE-A6 처리 시 농도 의존적으로 억제되는 것으로 나타났다. BGE-A6의 피부보습 관련 단백질 발현에 미치는 영향을 western blot으로 측정한 결과 HAS2와 AQP3 발현량 및 EGFR의 인산화를 처리 시 농도 의존적으로 증가시키는 것으로 나타났다. 결론적으로 피부세포에 H₂O₂를 처리하여 산화적 스트레스를 유도하였을 때 BGE-A6는 항산화 효소의 발현을 통해 세포보호 효과를 나타내고, AQP3, HAS2 및 콜라겐 단백질의 발현은 증가시키고 MMP-1의 발현을 감소시켜 피부보습 및 피부장벽 기능을 유지하는 것으로 사료된다. 또한, AQP3의 증가는 피부조직 및 피부세포막에 발현되는 EGF 수용체의 인산화를 증가시킴으로써 피부 건조에 대한 보호 효과도 있는 것으로 판단된다. 따라서 흑삼 추출물은 항산화 효과, 피부보습 및 장벽기능 유지를 통한 항노화 화장품 소재로 활용이 가능할 것으로 기대된다. In this study, black ginseng extract steamed six times at 95°C (BGE-A6) was evaluated for its antioxidant, anti-wrinkle, and skin moisturizing effects in human keratinocytes. When steamed six times at 95°C, the content of total ginsenosides including minor ginsenosides (Rg3, Rh4, Rk1, and Rg5) was the highest at 14.97 mg/g. The antioxidant properties of BGE-A6 were measured by using different in vitro antioxidant assays such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid, ABTS), and superoxide anion radical scavenging activity. The antioxidant activity of BGE-A6 was increased in a dose-dependent manner. A 2′,7′-dichlorofluorescin diacetate (DCFDA) assay and a western blot assay were performed to verify the reactive oxygen species (ROS) scavenging activity of BGE-A6 in H₂O₂-induced human keratinocytes. Pretreatment with 100 μg/mL of BGE-A6 inhibited intracellular ROS production by about 40%. Also, the expression of heme oxygenase-1 (HO-1), catalase (CAT), and superoxide dismutase-1 (SOD-1), very well-known as antioxidant enzymes, was increased by BGE-A6 in a dose-dependent manner. BGE-A6 was shown to significantly inhibit the expression of matrix metalloproteinase-1 and increase the production of type I collagen, thus confirming its anti-wrinkle activity (P<0.05). Furthermore, BGE-A6 significantly increased the phosphorylation of the epidermal growth factor receptors (EGFR) and the expression of moisturizing proteins, such as hyaluronic acid synthase 2 (HAS2) and aquaporin 3 (AQP3), indicating that it is effective in enhancing skin hydration. The results of this study suggest that BGE-A6 has potential for use as an effective compound in anti-aging cosmetics to address wrinkle formation and skin hydration.

40세 이하 관상동맥 질환자에서 장기적 임상 관찰

이승욱,정명호,박종철,박형욱,조상희,김원,김계훈,이상현,강경태,조장현,김남호,김건형,안영근,조정관,박종춘,강정채 全南大醫大 2001 전남의대학술지 Vol.37 No.2

서울의 Penicillinase Producing Neisseria gonorrhoeae 발생빈도(1998)

김재홍,김준호,반재용,이정우,황성주,정준규,정성태,강진문,조흔정,홍창의,정혜신,이한승,김이선,이봉길,이종호,선영우,한기덕,윤성필,이성훈,안종성,박석범,문승현,조항래,김형섭,류지호,황재영,박준홍,손상욱 한양대학교 의과대학 2001 한양의대 학술지 Vol.21 No.1

In recent years, gonorrhea has been pandemic and remains one of the most common STDs in the world, especially in developing countries. For the detection of a more effective therapeutic regimen and assessing the prevalence of Penicillinase Producing Neisseria gonorrhoeae(PPNG), we have been trying to study the patients who have visited the Venereal Disease Clinic of Choong-Ku Public Health Center in Seoul since 1980 by menas of the chromogenic cephalosporin method. In 1998, 93 strians of N. genorrhoeae were isolated, among which 60(64.5%) were PPNG. The prevalence of PPNG in Seoul, which had been decreased to 39% in 1996 after a peak of 74.3% in 1993, is increased to 64.5% in 1998.