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Role of Ca2+ in diallyl disulfide-induced apoptotic cell death of HCT-15 cells
Eun-KyungPark,권강범,Kie-InPark,Byung-HyunPark,Eun-ChungJhee 생화학분자생물학회 2002 Experimental and molecular medicine Vol.34 No.3
Diallyl disulfide (DADS) induced apoptosis throughthe caspase-3 dependent pathway in leukemia cellswas earlier reported from this laboratory. In thisstudy, we investigated the involvement of Ca2+ inDADS-induced apoptotic cell death of HCT-15,human colon cancer cell line. DADS induced the ele-vation of cytosolic Ca2+ by biphasic patern; rapidCa2+ peak at 3 min and following slow and sustainedelevation till 3 h after the addition of DADS. Produc-tion of H2O2 was also observed with its peak value at4 h. Apoptotic pathways including the sequence ofcaspase-3 activation, poly(ADP-ribose) polymerasecleavage, and DNA fragmentation by DADS werecompletely blocked by various inhibitors such asspecific caspase-3 inhibitor, free radical scavenger,and intracellular Ca2+ chelator. N-acetylcystein andcatalase treatment prevented the accumulation ofH2O2 and later caspase-3 dependent apoptotic path-way. However, these radical scavengers did notblock the elevation of intracellular Ca2+. Treatment of cells with 1,2-bis(2-aminophenoxy-ethane)-N,N,N-tetraacetic acid tetrakis -acetoxymethylester (BAPTA-AM), cellular Ca2+ chelator, resulted in acomplete blockage of the caspase-3 dependent apop-totic pathway of HCT-15 cells. It abolished the elevationof intracellular Ca2+, and furthermore, completely inhib-ited the production of H 2O2. These results indicate thatcytosolic Ca2+ elevation is an earlier signaling event inapoptosis of HCT-15 cells. Collectively, our data dem-onstrate that DADS can induce apoptosis in HCT-15cells through the sequential mechanism of Ca2+homeostasis disruption, accumulation of H 2O2, andresulting caspase-3 activation.
Fecal Metabolic Activities of Herbal Components to Bioactive Compounds
Dai-SikLee,Young-SukKim,Chang-NamKo,Ki-HoCho,Hyung-SupBae,Kyung-SupLee,Jung-JinKim,Eun-KyungPark 대한약학회 2002 Archives of Pharmacal Research Vol.25 No.3
The herbal components should be transformed to bioactive compounds by the intestinal bacteria and then expressed the pharmacological action of herbal medicines. Human fecal enzyme activities related to the metabolism of herbal components were measured. The metabolic activities of puerarin, poncirin, glycyrrhizin, ginsenoside Rb1 and ginsenoside Rb2 to their bioactive compounds were 3.51.18, 333.1183.64, 95.7107.1, 28.610.32 and 20.813.3 mmol/ h/g, respectively. The profile of these metabolic activities of glycyrrhizin and ginsenosides were not changed even if herbal extracts, water extract of Glycyrrhizae Radix and Ginseng Radix, instead of the isolated compounds were used. All the enzyme activities tested were not different between male and female, and between ages. However, the difference of these enzyme activities in individuals was significant. These results suggest that the metabolic activity of herbal components to bioactive compounds may be a factor of constitutional classification, and could be available for constitutional classifications, if the constitutional herbal medicines were used.