• KSBB Journal
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• 제22권3호
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• pp.168-173
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• 2007

형광을 이용한 암 진단을 위해 배양된 정상 및 암세포주에 광민감제인 5-ALA, ALA-methyl ester를 투여하고 세포 내 외에서 생성된 Protoporphyrin IX (PpIX)의 형광을 측정하여 5-ALA, ALA-methyl ester 투여의 최적 농도를 조사하였다. 정상 간세포주 (Chang) 및 간암 세포주 (HepG2)에 5-ALA와 ALA-methyl ester를 농도별로 투여하여 5-ALA와 ALA-methyl ester에 의해 유도된 PpIX의 생성을 확인하고, MTT assay로 세포생존율을 측정하였다. 배양된 cell에 5-ALA와 ALA-methyl ester를 투여한 후 24시간 동안 배양함으로써 생성되는 PpIX의 양은 형광의 강도로 측정하였다. 이 때의 형광 (emission) 스펙트럼은 여기 파장이 410 nm일 때 603.2 nm, 660.5 nm와 603.2 nm 및 661.4에서 형광 봉우리가 관찰되었다. PpIX의 형광 강도를 측정한 결과, PpIX는 정상세포에서는 낮은 농도로 축적이 되는 반면에 암세포에서 더 높은 농도로 축적되었으며, 세포 외보다는 세포 내에서 더 높은 농도로 축적됨을 알 수 있었다. 또한 5-ALA 및 ALA-methyl ester에 의한 PpIX의 생성에 대한 형광 강도는 ALA-methyl ester가 5-ALA보다 더 큰 것으로 관찰되었다.

• KSBB Journal
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• 제21권3호
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• pp.171-174
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• 2006

형광을 이용한 암 진단을 위해 배양된 정상 및 암세포주에 광민감제인 5-ALA를 투여하고 세포 내 외에서 생성된 Protoporphyrin IX(PpIX)의 형광을 측정하여 5-ALA 투여의 최적농도를 조사하였다. 정상 간세포주(Chang) 및 자궁경부암 세포주(HeLa)에 5-ALA를 농도별로 투여하여 ALA에 의해 유도된 PpIX의 생성을 확인하고, MTT assay로 세포생존율을 측정하였다. 배양된 cell에 5-ALA를 투여한 후 24시간 동안 배양함으로써 생성되는 PpIX의 양은 형광의 강도로 측정하였다. HeLa 세포주에 대한 5-ALA의 최적농도는 $50{\mu}g/ml$이며, 이 때의 형광(emission) 스펙트럼은 여기 파장이 410 nm일 때 602.3 nm, 659.9 nm에서 형광 봉우리가 관찰되었다. PpIX의 형광 강도를 측정한 결과, PpIX는 정상세포에서는 낮은 농도로 축적이 되는 반면에 암세포에서 더 높은 농도로 축적되었으며, 세포 외보다는 세포 내에서 더 높은 농도로 축적됨을 알 수 있었다.

• KSBB Journal
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• 제22권2호
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• pp.67-72
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• 2007

형광을 이용한 암 진단을 위해 배양된 정상 폐세포 및 폐암 세포주에 광민감제인 5-ALA를 투여하고 세포 내 외에서 생성된 protoporphyrin IX (PpIX)의 형광을 측정하여 5-ALA 투여의 최적 농도를 조사하였다. 정상 폐세포주 (Hel299) 및 폐암 세포주 (A549, NCI-H460)에 5-ALA를 $0\sim800{\mu}g/mL$ 농도별로 투여하여 24시간 동안 배양한 다음 MTT assay로 세포증식 저해율 및 이때 생성되는 PpIX의 양을 형광의 강도로 측정하였다. 그 결과 Hel299 및 A549에서는 5-ALA의 처리농도가 증가할수록 세포 증식의 저해율이 증가하였으나 NCI-H460에서는 세포 증식이 저해되지 않았다. 그리고 폐암세포인 A549와 NCI-H460에 대한 5-ALA의 최적농도는 $100{\mu}g/mL$이며, 이때의 형광 (emission) 스펙트럼은 여기 파장이 410 nm일 때 세포 외에서는 615.8 nm와 660.8 nm, 616.7 nm와 660.2 nm, 세포 내에서는 603.2 nm와 661.4 nm, 603.5 nm와 661.4 nm에서 각각 형광 봉우리가 관찰되었다. 또한 PpIX를 형광 강도로 측정하면, PpIX는 정상세포에서는 낮은 농도로 축적이 되는 반면에 암세포에서 높은 농도로 축적되었으며, 세포 외보다는 세포 내에서 더 높은 농도로 축적됨을 알 수 있었다.

• Bulletin of the Korean Chemical Society
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• 제28권1호
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• pp.129-132
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• 2007

• Journal of Photoscience
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• 제9권2호
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• pp.512-514
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• 2002

S-aminolevulinic acid (ALA) is a new kind drug used in photodynamic therapy. ALA-PDT have successfully used in superficial malignancies and some skin diseases. Here the effects of ALA-PDT were studied on leukemia cells and hepatoma cells to explore the application on different kind cancers. It was found from the fluorescence emission spectra, that after ALA incubation the sensitizer - protoporphyrin IX (PpIX) was endogenously produced in both leukemia and hepatoma cells. The fluorescence images showed that the PpIX distribute in cytoplasm. However the efficiency of ALA photodynamic inactivation to two cell lines was different. The leukemia cells were more sensitive for ALA-PDT than hepatoma cells, revealing that the ALA-PDT effect is cell line dependent. However by using ALA-Hexyl ester (He-ALA) instead of ALA, the cell photo-inactivation was improved. The PDT efficiency of He-ALA was 10 times high than that of ALA, showing He-ALA is a very promising drug in ALA-PDT.

• 동의생리병리학회지
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• 제22권4호
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• pp.871-877
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• 2008

Heme oxygenase-1 (HO-1), an inducible heme-degrading enzyme, is expressed by macrophages and endothelial cells in response to various stresses and mediators of inflammation. HO-1 has been recently implicated in regulation of angiogenesis via expression of VEGF. The purpose of this study was to determine the effects of HO-1 modulation on the collagen-induced arthritis (CIA) model and on angiogenesis via up- regulation of VEGF expression in human synovial fibroblast. DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 1 to day 35 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, histopathologic changes were examined on the joints. VEGF expression in paws were measured by immunohistochemical stain. mRNA expression of HO-1 and VEGF stimulated with various concentration of $TNF-{\alpha}$, CoPP accessed on human synovial fibroblast by RT-PCR. Effects of pretreatment with SnPP on mRNA expression of HO-1 and VEGF in the presence of CoPP and $TNF-{\alpha}$ in synovial fibroblast was accessed by Real-time RT-PCR. Administration of cobalt protoporphyrin IX significantly induced the inflammatory response, with increased arthritis index and expression of VEGF in the paws of the arthritis models. Treatment with SnPP significantly reduced the severity of CIA through inhibition of joint inflammation and cartilage destruction. The expression of VEGF were also significantly reduced by SnPP treatment in the paw. CoPPIX as inducer of HO-1, increased HO-1 and VEGF expression dose dependently in synovial fibroblast. In contrast, inhibition of HO-1 activity by SnPPIX abrogated CoPPIX-induced HO-1 and VEGF production in synovial fibroblast. Stimulation with $TNF-{\alpha}$ increased HO-1 and VEGF expression itself and showed additive effect on HO-1 and VEGF expression when it treated with CoPP. When SnPP was treated with CoPP and $TNF-{\alpha}$, it abrogated the CoPP induced HO-1 and VEGF expression and also abrogated $TNF-{\alpha}$ induced HO-1 and VEGF expression in synovial fibroblast. The effects of HO-1 induction in rheumatoid arthritis results in aggravation of arthritis via up-regulation of VEGF. I concluded that inhibition of the expression or activity of HO-1 could be a therapeutic target of rheumatoid arthritis.

• Journal of Photoscience
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• 제9권2호
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• pp.521-523
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• 2002

5-aminolevulinic acid (ALA) has been used to stimulate endogenous protoporphyrin IX (PpIX) in tumor and then initiate PDT. Recently, ALA-Hexyl ester (He-ALA) was found much effective than ALA on producing PpIX in cancer cells. To clarify the transportation mechanism of ALA and He-ALA, the detection of them is the important step. ALA and its derivatives all don't emit fluorescence, so the Raman spectroscopy was used here for the direct detection of ALA and He-ALA. The results showed that ALA and He-ALA have the common strong Raman peaks at 2930, 2950 CM$\^$-1/, due to the CH$_2$ vibration. The peak 3050 CM$\^$-1/ appeared in ALA spectrum can be attributed to OH vibration, while the peaks of 2860, 2900 CM$\^$-1/ in He-ALA spectrum were assigned as the modes of CH$_3$. This Raman spectral characteristic is consistence with the structure difference of He-ALA and ALA. Thus, Raman spectroscopy provides a new way to detect and distinguish ALA and He-ALA, and could be explored further in biology system.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1999년도 학술발표회 진행표 및 논문초록
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• pp.44-44
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• 1999

Yeast cytochrome c peroxidase (CcP) was cloned and overexpressed in E. coli, and purified by a Ni$^{2＋}$-affinity column. HoloCcP was obtained by reconstituting apoCcP with Mn$^{3+}$-protoporphyrin IX (MnPP). Electron paramagnetic resonance (EPR) spectra of spin-labeled holoCcP showed a slightly more immobilized signal than spin-labeled apoCcP.(omitted)

• The Korean Journal of Physiology and Pharmacology
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• 제15권2호
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• pp.83-88
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• 2011

A large body of evidence has indicated that induction of endogenous antioxidative proteins seems to be a reasonable strategy for delaying the progression of cell injury. In our previous study, cilostazol was found to increase the expression of the antioxidant enzyme heme oxygenase-1 (HO-1) in synovial cells. Thus, the present study was undertaken to examine whether cilostazol is able to counteract tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced cell death in endothelial cells via the induction of HO-1 expression. We exposed human umbilical vein endothelial cells (HUVECs) to TNF-${\alpha}$ (50 ng/ml), with or without cilostazol ($10{\mu}M$). Pretreatment with cilostazol markedly reduced TNF-${\alpha}$-induced viability loss in the HUVECs, which was reversed by zinc protoporphyrine IX (ZnPP), an inhibitor of HO-1. Moreover, cilostazol increased HO-1 protein and mRNA expression. Cilostazol-induced HO-1 induction was markedly attenuated not only by ZnPP but also by copper-protoporphyrin IX (CuPP). In an assay measuring peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$) transcription activity, cilostazol directly increased PPAR-${\gamma}$ transcriptional activity which was completely abolished by HO-1 inhibitor. Furthermore, increased PPAR-${\gamma}$ activity by cilostazol and rosiglitazone was completely abolished in cells transfected with HO-1 siRNA. Taken together, these results indicate that cilostazol up-regulates HO-1 and protects cells against TNF-${\alpha}$-induced endothelial cytotoxicity via a PPAR-${\gamma}$-dependent pathway.

• 동의생리병리학회지
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• 제18권4호
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• pp.1071-1077
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• 2004

Mudanpi (Cortex Moutan Radicis; the root cortex of Paeonia suffruticosa Andrews) is an important Chinese crude drug used in many oriental prescriptions. 1,2,3,4,6-Penta-O-galloyl-beta-D-glucose (PGG), a major component of this crude drug, has been shown to possess potent antioxidant, anti-mutagenic and anti-proliferative effects. In this study, I examined whether PGG could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 expression and HO activity. Exposure of Neuro 2A cells to PGG (10-50μM) resulted in a concentration- and time-dependent induction of HO-1 mRNA, and protein expressions and heme oxygenase activity. PGG protected the cells from hydrogen peroxide-induced cell death. The protective effect of PGG on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX (ZnPP IX), a HO inhibitor. These results indicate that PGG is a potent inducer of HO-1 and HO-1 induction is responsible for the PGG-mediated cytoprotection against oxidative damage.