Artery Relaxation by Chalcones Isolated from the Roots of Angelica keiskei

136

Six chalcones from Angelica keiskei KOIDZUMI (Ashitaba in Japanese) and two chalcones from Humulus lupulus L. (hop) were examined for their cytotoxicity in two human neuroblastoma cell lines (IMR-32 and NB-39) and normal cells (primary culture of rat cerebellar granule cells) by [3-(4,5)-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. All chalcones exhibited cytotoxicity against neuroblastoma cells, and two of them (isobavachalcone and xanthoangelol H) had no effect on normal cells even at high concentration (10 ؊4 M) exposure. Typical morphologic features of apoptosis, including cell shrinkage, chromatin condensation, nuclear fragmentation and formation of apoptotic bodies, were observed in isobavachalcone-treated cells by Hoechst 33342 staining. Western blot analysis showed that isobavachalcone significantly reduced pro-caspase-3 and pro-caspase-9, and subsequently increased the level of cleaved caspase-3 and cleaved caspase-9 in both neuroblastoma cell lines. Moreover, Bax was markedly induced by isobavachalcone application. These results suggest that isobavachalcone induces apoptotic cell death in neuroblastoma via the mitochondrial pathway and has no cytotoxicity against normal cells. Therefore, isobavachalcone may be applicable as an efficacious and safe drug for the treatment of neuroblastoma.

Section: )mentioning

confidence: 99%

Isobavachalcone, a Chalcone Constituent of Angelica keiskei, Induces Apoptosis in Neuroblastoma

Nishimura

Tabata

Arakawa

et al. 2007

136

“…Recent studies have also indicated that the extracted chalcones from the roots of Angelica keiskei KOIDZUMI have beneficial effects including vasodilatation, antitumor and antimetastatic activities. [17][18][19] The purpose of the present study is to evaluate whether chalcones extracted from Angelica keiskei KOIDZUMI have an inhibitory effect on NF-kB activation. To attain this, we examined the effects of xanthoangelol, xanthoangelol D, E, and F on NF-kB activation in PAECs.…”

Section: )mentioning

confidence: 99%

Xanthoangelol D Isolated from the Roots of Angelica keiskei Inhibits Endothelin-1 Production through the Suppression of Nuclear Factor-.KAPPA.B

Sugii

Ohkita

Taniguchi

et al. 2005

Self Cite

Many vascular diseases are closely related to inflammatory processes, where activation of the transcription factor nuclear factor-kappa B (NF-kB) plays a pivotal role by augmenting the expression of various genes such as cytokines, chemokines, leukocyte-adhesion molecules.1,2) NF-kB is found in the cytoplasm of most cells as an inactive complex bound to an inhibitory protein, inhibitor kappa B (IkB). It is also well known that treatment with a variety of stimuli including tumor necrosis factor-a (TNF-a) and reactive oxygen species (ROS) results in the rapid activation of NF-kB, through the phosphorylation of IkB and its subsequent proteolytic degradation by the proteasome-dependent pathway. 2)Recent studies have shown that the inhibition of gene expressions through NF-kB activation is one of the favorable treatments in vascular and inflammatory diseases. 3,4) Endothelin-1 (ET-1) is a potent vasoconstrictor peptide purified from the supernatant of cultured porcine aortic endothelial cells (PAECs) 5) and possesses a number of biological activities leading to vascular disorders. 6) ET-1 biosynthesis and release appear to be regulated at the transcriptional level because ET-1 release from endothelial cells (ECs) is constitutive. Several studies have indicated that various substances such as thrombin, 7) transforming growth factor-b1, 8) and TNF-a 9) stimulate the ET-1 gene expression in ECs by DNA binding of transcription factors such as activator protein-1 (AP-1) and nuclear factor-1 (NF-1). Recent studies have shown that NF-kB is also one of modulators in ET-1 production. [10][11][12] Chalcones are obtained from various natural plant sources. It has been reported that chalcones possess various biological properties, including anti-inflammatory, analgesic, antioxidant, antibacterial, and antifungal activities. 13,14) Our previous studies have demonstrated that the roots of Angelica keiskei KOIDZUMI are abundant in chalcones such as xanthoangelols and 4-hydroxyderricin. 15,16) These roots have traditionally been used as a health food with diuretic, laxative, analeptic and galactagogic effects. Recent studies have also indicated that the extracted chalcones from the roots of Angelica keiskei KOIDZUMI have beneficial effects including vasodilatation, antitumor and antimetastatic activities. [17][18][19] The purpose of the present study is to evaluate whether chalcones extracted from Angelica keiskei KOIDZUMI have an inhibitory effect on NF-kB activation. To attain this, we examined the effects of xanthoangelol, xanthoangelol D, E, and F on NF-kB activation in PAECs. In addition, we evaluate the effects of xanthoangelols on ET-1 production in PAECs. MATERIALS AND METHODSMaterials Xanthoangelol D, E, F and xanthoangelol were isolated from the roots of Angelica keiskei KOIDZUMI by Baba et al. (Fig. 1)

“…An equal volume of stop solution (0.04 N HCl in isopropanol) to that of the culture medium was then added to each well and the absorbance at 570 nm (peak) and 630 nm (bottom) was ; Funabashi, Japan: c Department of Materials and Applied Chemistry, College of Science and Technology, Nihon University;[1][2][3][4][5][6][7][8] Japan: and d Department of Pediatrics, School of Medicine, Nihon University; 30-1 Oyaguchikami-cho, Itabashi-ku, Tokyo 173-0032, Japan. Received February 28, 2005; accepted April 25, 2005 Xanthoangelol, a major chalcone constituent of the stem exudates of Angelica keiskei, was evaluated for cell toxicity and apoptosis-inducing activity in human neuroblastoma (IMR-32) and leukemia (Jurkat) cells.…”

Section: -[45-dimethylthiazol-2-yl]-25-diphenyltetrazolium Bromidementioning

confidence: 99%

Xanthoangelol, a Major Chalcone Constituent of Angelica keiskei, Induces Apoptosis in Neuroblastoma and Leukemia Cells

Tabata

Motani

Takayanagi

et al. 2005

105

Neuroblastoma is the most common solid pediatric tumor and remarkable for its clinical heterogeneity. Despite recent advances in chemotherapy, the prognosis of advanced neuroblastoma is still very poor. However, some favorable types of neuroblastoma, especially in infants under 1 year of age, are known to regress spontaneously or mature even if widespread metastases to bone marrow, skin and/or liver (special stage: stage IVS) are present. Apoptosis is known to occur in normal development of nervous systems, and neuroblastoma is generated from neural crest cells when the apoptotic systems do not carry out. Delayed implementation of the normal apoptotic pathway has been proposed as an explanation for the spontaneous regression of favorable neuroblastoma.1) It is reported that resistance to apoptosis plays a contributory role in the mechanism of the aggressive behavior shown by advanced neuroblastoma.2) Acute lymphocytic leukemia, like advanced neuroblastoma, is also a pediatric disease that is difficult to treat, especially in older children or those with a high amount of leukemic cells in the peripheral blood.Angelica keiskei has been used traditionally in Japan as a diuretic, laxative, analeptic and galactagogue, and an A. keiskei extract was previously reported to affect metabolic activity 3,4) and vasoconstriction 5) in rats. Moreover, A. keiskei and a major chalcone constituent of this plant, xanthoangelol, reportedly have inhibitory effects against tumor promoter activity 6,7) and metastasis. 8) Xanthoangelol possesses a chalcone structure, and some compounds related to calchones are known to have antitumor activity and to induce apoptosis. Quercetin chalcone was reported to reduce the size of implanted colon-25 tumors in vivo. 9) However, there has been no report on the effects of chalcones, including xanthoangelol, on neuroblastoma.In this study, we examined the antitumor effect and apoptosis-inducing activity of xanthoangelol against a human neuroblastoma cell line (IMR-32), and also a leukemia cell line (Jurkat) which have been widely used in previous studies of apoptosis. MATERIALS AND METHODS Materials Xanthoangelol(3Ј-C-geranyl-2Ј,4,4Ј-trihydroxychalcone) was isolated from the stem exudate of A. keiskei 6) and dissolved in dimethyl sulfoxide (DMSO) (final concentration 0.2%). IMR-32 and Jurkat were maintained in RPMI-1640 medium (Invitrogen) supplemented with 100 U/ml penicillin, 100 mg/ml streptomycin and 10% fetal bovine serum (FBS) (Invitrogen). The cells were maintained at 37°C/5% CO 2 in a humid environment.Trypan Blue Exclusion Assay The cells ( 1ϫ10 6 ) were plated into a 60-mm dish and maintained for 24 h. Xanthoangelol (final concentrations 10 Ϫ6 , 10 Ϫ5, 10 Ϫ4 M) and vehicle were applied for 48 h. For the IMR-32 cell protocol, the cells were stripped using 0.05% trypsin-EDTA solution after washing them in phosphate-buffered saline (Ϫ). They were then washed in RPMI-1640 medium (with 10% FBS) and counted with a phase-contrast microscope immediately after addition of an equal volume of 1% tr...