• KSBB Journal
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• v.22 no.3
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• pp.168-173
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• 2007

To clarify the usefulness of fluorescent diagnosis for cancer, we investigated the optimal method of administrating 5-aminolevulinic acid (5-ALA), 5-aminolevulinic acid methyl ester (ALA-methyl ester) by analyzing fluorescence signal of Protoporphyrin IX (PpIX) in the cultured normal and cancer cells. 5-ALA and ALA-methyl ester was injected as a photosensitizer to the cancer liver cells (HepG2) and normal liver cells (Chang). Chang and HepG2 cells were incubated with various concentrations of 5-ALA and ALA-methyl ester (0-800 ${\mu}g/mL$). The accumulation of PpIX induced by 5-ALA and ALA-methyl ester was in HepG2 and Chang. The cell viability was measured by MTT assay. Fluorescence of PpIX in HepG2 cell was excited at a wavelength ($\lambda$ = 410 nm) and showed an emission spectrum at 603.2 nm, 660.8 nm and 603.2 nm, 661.4 nm which could be related to the PpIX generation induced by the applied 5-ALA and ALA-methyl ester, respectively. The experimental results showed that fluorescence signal of PpIX was proportional to the concentration of 5-ALA and ALA-methyl ester in tumor cells, but measured with low concentration in normal cells. Another results showed that the PpIX formation rate induced by ALA-methyl ester is higher than that of 5-ALA.

• KSBB Journal
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• v.21 no.3
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• pp.171-174
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• 2006

To clarify the usefulness of fluorescent diagnosis for cancer, We investigated the optimal method of administrating 5-aminolevulinic acid(5-ALA) by analyzing fluorescence signal of Protoporphyrin IX(PpIX) in the cultured normal and cancer cells. 5-ALA was injected as a photosensitizer to the cervico-uterine cancer cell line(HeLa) and in normal liver cells(Chang). Chang and HeLa cells were incubated with various concentrations of 5-ALA($0-800{\mu}g/ml$). The accumulation of PpIX induced by 5-ALA was in HeLa and Chang cells. The cell viability was measured by MTT assay. The optimal concentration of ALA that induced maximum levels of PpIX was $50{\mu}g/ml$ in HeLa cell cultured for 24 hours after 5-ALA injection. Fluorescence of PpIX in HeLa cell was excited at a wavelength(${\lambda}$=410 nm) and showed an emission spectrum at 602.3 nm, 659.9 nm which could be related to the PpIX generation induced by the applied 5-ALA. The experimental results showed that fluorescence signal of PpIX was proportional to the concentration of 5-ALA in cancer cells, but measured with low concentration in normal cells.

• KSBB Journal
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• v.22 no.2
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• pp.67-72
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• 2007

This study investigates the optimal method of administrating 5-aminolevulinic acid (5-ALA) in the context of fluorescence detection by analyzing protoporphyrin IX (PpIX) fluorescence in the cultured normal and cancer cells. 5-ALA was injected as a photosensitizer to the lung cancer cells (A549, NCI-H460) and normal lung cells (HeI299). Hel299, A549, and NCI-H460 cells were incubated with various concentrations of 5-ALA ($0\sim800{\mu}g/mL$). The accumulation of PpIX induced by 5-ALA was observed in A549, NCI-H460 and Hel299 cells. The cell viability was estimated by means of the MTT assay. Formation of PpIX was measured by fluorescence spectroscopy. Especially, formation of PpIX in cancer cells was higher than normal cells. This study suggests that the difference of PpIX induced in normal and cancer cells treated with 5-ALA may use by means of fluorescence diagnosis for cancer.

• Bulletin of the Korean Chemical Society
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• v.28 no.1
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• pp.129-132
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• 2007

• Journal of Photoscience
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• v.9 no.2
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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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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• v.9 no.2
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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.

• Proceedings of the Korean Biophysical Society Conference
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• 1999.06a
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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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• v.15 no.2
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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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.