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http://dx.doi.org/10.5352/JLS.2014.24.3.242

Isolation and Identification of a Photosensitizer from Pueraria thunbergiana Leaves that Induces Apoptosis in SK-HEP-1 Cells  

Lee, Jun Young (Department of Life Science & Environmental Biochemistry, Pusan National University)
Kim, Mi Kyeong (Department of Life Science & Environmental Biochemistry, Pusan National University)
Ha, Jun Young (Department of Life Science & Environmental Biochemistry, Pusan National University)
Kim, Yong Gyun (Department of Life Science & Environmental Biochemistry, Pusan National University)
Hong, Chang Oh (Department of Life Science & Environmental Biochemistry, Pusan National University)
Kim, So Young (Department of Convergence Medicine and Pharmaceutical Biosciences, Graduate School, Chung-Ang University)
Kim, Chung-Hwan (Department of Clinical Pathology, Masan University)
Kim, Keun Ki (Department of Life Science & Environmental Biochemistry, Pusan National University)
Publication Information
Journal of Life Science / v.24, no.3, 2014 , pp. 242-251 More about this Journal
Abstract
The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.
Keywords
Anticancer; apoptosis; photosensitizer; Pueraria thunbergiana; SK-HEP-1;
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Times Cited By KSCI : 5  (Citation Analysis)
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