Induction of apoptosis by carbazole alkaloids isolated from Murraya koenigii.
In the current study, we isolated 10 carbazole alkaloids from the plant species Murraya koenigii (Rutaceae), and examined their effects on the growth of the human leukemia cell line HL-60. Three carbazole alkaloids, mahanine (6), pyrayafoline-D (7) and murrafoline-I (9), showed significant cytotoxicity against HL-60 cells. Fluorescence microscopy with Hoechst 33342 staining revealed that the percentage of apoptotic cells with fragmented nuclei and condensed chromatin was increased in a time-dependent manner after treatment with each alkaloid. Interestingly, each carbazole alkaloid induced the loss of mitochondrial membrane potential. In addition, both caspase-9 and caspase-3 were also time-dependently activated upon treatment with the alkaloids. Caspase-9 and caspase-3 inhibitors suppressed apoptosis induced by these alkaloids. The results suggest that these three alkaloids induced apoptosis in HL-60 cells through activation of the caspase-9/caspase-3 pathway, through mitochondrial dysfunction.
Ito
C
C
Department of Medicinal Chemistry, Faculty of Pharmacy, Meijo University, Nagoya, Japan.
Itoigawa
M
M
Nakao
K
K
Murata
T
T
Tsuboi
M
M
Kaneda
N
N
Furukawa
H
H
D017209:Apoptosis; D002227:Carbazoles / Q000737:chemistry* / Q000494:pharmacology*; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D051336:Mitochondrial Membranes / Q000187:drug effects; D032101:Murraya / Q000737:chemistry*; D009842:Oligopeptides / Q000494:pharmacology; D010946:Plants, Medicinal / Q000737:chemistry*; D013997:Time Factors; D018675:Toxicity Tests
D002227:Carbazoles; D009842:Oligopeptides; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; C403754:benzyloxycarbonyl-leucyl-glutamyl-histidyl-aspartic acid fluoromethyl ketone; C505407:CASP3 protein, human; C506433:CASP9 protein, human; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases