The cytotoxic and antiproliferative effects of gamavuton-0 in rat basophilic Leukemia cells

Gamavuton-0 (GVT-0), also named as 1,5-bis(4’-hydroxy-3’- methoxyphenyl)-1,4-pentadiene-3 one is a 1,5-diphenyl-1,4-pentadiene-3-one analog of curcumin by modifying the center site of curcumin leading to 1,4-pentadiene-3-ones to maintain the hydroxy moiety at aromatic rings which are responsible for its biological activities. Curcumin has been reported to have potent anticarcinogenic effects. Besides, curcumin was found to induce apoptosis in human Leukemia cells. In our study, we investigated the cytotoxic and antiproliferative effects of gamavuton-0 in rat basophilic leukemia cells. Cell viability was determined by WST-1 assay. In brief, tetrazolium salts were cleaved to formazan by cellular enzymes of viable cells, determined by colorimetric methods with a microplate (ELISA) reader at 450 nm.In the present study, we evaluated cytotoxic and proliferative effects of GVT-0 in rat basophilic leukemia cells. In the study, GVT-0 induced rat basophilic leukemia cells death in a dose dependent manner after overnight incubation. GVT-0 also impaired the content of histamine and b-hexoaminidase enzyme in cells. However, the cytotoxic effect of GVT-0 (IC50 : 43,67 mM) was less potent than this of curcumin (IC50 : 29,14 mM). GVT-0 (1 mM) also showed a significant inhibition of cell growth after 48 and 72 hr. Its fact indicates that GVT-0 could prolong the cells doubling time. These results provide useful information to guide the development of new synthetic compounds for the treatment of cancer diseases.Key words : gamavuton-0, curcumin, cancer, cytotoxic, antiproliferative

The effects of PGV-1 and PGV-2 on the b-hexosaminidase release from intraceluller calcium ion-induced mast cells

PGV-1 or 2,5-bis(4'-hydroxy-3',5'-dimethylbenzylidene)cyclopentanone and PGV-2 or 2,5-bis(4'-hydroxy-3',5'-diethylbenzylidene)cyclopentanone are two benzylidene cyclopentanone analogues of curcumin. In our study, weinvestigated the effects of these compounds on the b-hexoaminidase enzyme release from mast cell culture (RBL-2H3 cell line). Thapsigargin and ionomycin were used as intracellular calcium ion stimulants for inducing b-hexoaminidase enzyme release from mast cells. The release of b-hexoaminidase enzyme was determined by colorimetric methods with substrate, p-nitrophenyl-2-acetamido-2-deoxy-b-D-glucopyranocide, and a microplate reader at 405 nm. In present study, treatment of 0.5 mM thapsigargin or 1 mM ionomycin could stimulate therelease of b-hexoaminidase enzyme from RBL-2H3 cells by 43.91 ± 1.30 % dan 52.93 ± 2.07 %, respectively. PGV-1 and PGV-2 showed inhibitory effects on the b-hexoaminidase enzyme release from RBL-2H3 cells induced by the increase of intraceluller calcium ion in dose-dependent manner. At the dose of 100 mM, PGV-1 and PGV-2, respectively, inhibited the b-hexoaminidase enzyme release by 73.51 ± 8.69 % and 66.42 ± 8.63 % on thapsigargin experiments; and by 89.73 ± 3.23 % and 38.57 ± 5.32 % on ionomycin experiments. The IC50 values of their effects on the b-hexoaminidase enzyme release from RBL-2H3 cells, respectively, were 22.20 mM and 22.27 mM on thapsigargin experiment; and 22.77 mM and >100 mM on ionomycin experiment. Based on the results, the inhibitory effect of PGV-1 and PGV-2 on the b-hexoaminidase enzyme release from RBL-2H3 cells involving mechanisms related to the alteration on activationprocesses of intracellular calcium ion on mast cells.Key words : Curcumin, PGV-1, PGV-2, mast cells, b-hexoaminidase enzym

Comparison of Cytotoxic and Antiproliferative Effects of Benzylidenecyclopentanone Analogues of Curcumin on RBL-2H3 Cells

Curcumin is a natural yellow pigment isolated from the rhizomes of Curcuma longa L. (turmeric), and has several pharmacological effects and no toxicity in both in animal and human clinical study. However, the problem of curcumin is its stability because of its active methylene moiety. Modification of this moiety to cyclopentanone is expected to increase the stability. Previous study reported that benzylidenecyclopentanone analogues of curcumin showed inhibitory effect on histamine release from RBL-2H3 (rat basophilic leukemia) cells, a tumor analog of mast cells. One of them, the hydroxy-methoxy analog (PGV-O), showed more potent effect than that of curcumin. In the present study, some benzylidenecyclopentanone analogues of curcumin were evaluated for their effects on th~ viability and proliferation of RBL-2H3 cells. Viable cells were counted under a light microscope with a cells-counting chamber or using the cell viability reagent W5T-1. The results showed that mast cell viability and histamine content were not affected by curcumin and benzylidene cyclopentanone for 30 min incubation, however, impaired for overnight incubation. The hydroxy-dimethyl benzylidene analog (PGV-l) strongly decreased the mast cells viability for overnight incubation, and its effect was highest among the other analogues. In the proliferation study, this compound also strongly inhibited the proliferation of mast cells, whereas curcumin and hydroxy-methoxy benzylidene analog inhibited the proliferation slightly. There were no inhibitory effects on mast cells proliferation treated by dibenzylidenedihydroxybenzylideneand hydroxy -diethy lbenzy lidene cyclopentanone

Inhibitory effect of THPGV-0 on the histamine release from antigen-induced RBL-2H3 cells

Tetrahydropentagamavunon-0  (THPGV-0)  is  assumed  to  be  main metabolite  product  of  biotransformation  process  of  PGV-0.  THPGV-0  was synthesized  by  converting  PGV-0  to  the  compound  by  hydrogenation  with  Pd/C as a catalyst. PGV-0 potently inhibited the histamine release from  rat mast cells in  vitro  and  in  vivo,  however,  ironically  only  traces  amount  of  compound  was found in the blood. THPGV-0 is assumed to have important roles in the biological effects of PGV-0 in vivo. In present study, we investigated the antiallergy effect of  THPGV-0  in  compare  to  this  of  PGV-0  in  vitro.  The  study  was  performed  by using  rat  basophilic  leukemia  (RBL-2H3)  cell  line,  a  tumor  analog  of  mucosal mast  cells.  DNP-BSA,  an  antigen,  was  used  as  an  inducer  for  stimulating  the histamine  release  from  mast  cells.  In  present  study,  THPGV-0  at  low concentration  did  not  succeed  to  inhibit  the  histamine  release,  but  at  higher concentration (30 and 100  M) showed strong effects. THPGV-0 at concentration of  100  M  depleted  the  histamine  release  by  96.10  0.51%.  In  compare  to PGV-0,  THPGV-0  has  higher  efficacy  but  less  potent.  In  the  study,  the possibilities  of  the  spontaneous  release  from  RBL-2H3  cells  by  the  compounds were also observed. All concentrations of THPGV-0 as well as PGV-0 showed low spontaneous histamine release, less than 10 % of the total histamine contained in RBL-2H3 cells.Key words: tetrahydropentagamavunon-0, allergy, histamine, RBL-2H3 cell

Comparison of Cytotoxic and Antiproliferative Effects of Benzylidenecyclopentanone Analogues of Curcumin on RBL-2H3 Cells

Curcumin is a natural yellow pigment isolated from the rhizomes of Curcuma longa L. (turmeric), and has several pharmacological effects and no toxicity in both in animal and human clinical study. However, the problem of curcumin is its stability because of its active methylene moiety. Modification of this moiety to cyclopentanone is expected to increase the stability. Previous study reported that benzylidenecyclopentanone analogues of curcumin showed inhibitory effect on histamine release from RBL-2H3 (rat basophilic leukemia) cells, a tumor analog of mast cells. One of them, the hydroxy-methoxy analog (PGV-0), showed more potent effect than that of curcumin. In the present study, some benzylidenecyclopentanone analogues of curcumin were evaluated for their effects on the viability and proliferation of RBL-2H3 cells. Viable cells were counted under a light microscope with a cells-counting chamber or using the cell viability reagent WST-1. The results showed that mast cell viability and histamine content were not affected by curcumin and benzylidene cyclopentanone for 30 min incubation, however, impaired for overnight incubation. The hydroxy-dimethyl benzylidene analog (PGV-1) strongly decreased the mast cells viability for overnight incubation, and its effect was highest among the other analogues. In the proliferation study, this compound also strongly inhibited the proliferation of mast cells, whereas curcumin and hydroxy-methoxy benzylidene analog inhibited the proliferation slightly. There were no inhibitory effects on mast cells proliferation treated by dibenzylidene; dihydroxybenzylidene; and hydroxy-diethylbenzylidene cyclopentanone.Keywords : viability, proliferation, curcumin, benzylidene cyclopentanone, RBL-2H3 cell

Comparison of Cytotoxic and Antiproliferative Effects of Benzylidenecyclopentanone Analogues of Curcumin on RBL-2H3 Cells

Curcumin is a natural yellow pigment isolated from the rhizomes of Curcuma longa L. (turmeric), and has several pharmacological effects and no toxicity in both in animal and human clinical study. However, the problem of curcumin is its stability because of its active methylene moiety. Modification of this moiety to cyclopentanone is expected to increase the stability. Previous study reported that benzylidenecyclopentanone analogues of curcumin showed inhibitory effect on histamine release from RBL-2H3 (rat basophilic leukemia) cells, a tumor analog of mast cells. One of them, the hydroxy-methoxy analog (PGV-0), showed more potent effect than that of curcumin. In the present study, some benzylidenecyclopentanone analogues of curcumin were evaluated for their effects on the viability and proliferation of RBL-2H3 cells. Viable cells were counted under a light microscope with a cells-counting chamber or using the cell viability reagent WST-1. The results showed that mast cell viability and histamine content were not affected by curcumin and benzylidene cyclopentanone for 30 min incubation, however, impaired for overnight incubation. The hydroxy-dimethyl benzylidene analog (PGV-1) strongly decreased the mast cells viability for overnight incubation, and its effect was highest among the other analogues. In the proliferation study, this compound also strongly inhibited the proliferation of mast cells, whereas curcumin and hydroxy-methoxy benzylidene analog inhibited the proliferation slightly. There were no inhibitory effects on mast cells proliferation treated by dibenzylidene; dihydroxybenzylidene; and hydroxy-diethylbenzylidene cyclopentanone. Keywords : viability, proliferation, curcumin, benzylidene cyclopentanone, RBL-2H3 cell

Effects of Flavonoids Isolated From Orange Jasmine (Murraya Paniculata [L.] Jack.) on Histamine Release From Mast Cells

Murraya paniculata [L.] Jack. (Kemuning) is a plant that grows widely in some areas of Indonesia. Studies related to this plant have been widely explored especially isolation of its active compounds. The plant contents several active compounds such as flanovoids. In the study, three flavonoid isolated from M. paniculata were evaluated for their effect on histamine release from mast cells (RBL-2H3 cells). These compounds were 3,3',4',5,5',6,7,8-octamethoxyflavone; 3,3',4',5,5',6,7-heptamethoxyflavone and 3, 3', 4', 5, 5', 7–hexamethoxyflavone. The histamine inducers used in the study were DNP24-BSA dan thapsigargin, inducing the histamine release immunologically and non-immunologically, respectively. In the study, heptamethoxyflavone and hexamethoxyflavone did not influence the histamine release from mast cells significantly. However, octamethoxyflavone could increase the histamine release from RBL-2H3 cells in absence and presence the histamine inducers. The flavanoid could increase the release of histamine up to 50 %. Based on the results, polymethoxy moieties at the structure of flavonoid have a significant role to emerge the histamine-release stimulating effect from mast cells

Effects of aegeline, a main alkaloid of aegle marmelos correa leaves, on the histamine release from mast cells.

Aegeline or N-[2-hydroxy-2(4-methoxyphenyl) ethyl]-3-phenyl-2-propenamide is a main alkaloid isolated from Aegle marmelos Correa collected in Yogyakarta Indonesia. In our study, we investigated the effects of aegeline on the histamine release from mast cell. The study was performed by using (1) rat basophilic leukemia (RBL-2H3) cell line, and (2) rat peritoneal mast cells (RPMCs). DNP24-BSA, thap pe of mast cell and also involved some mechanisms related to intracellular Ca 2+ signaling events via the same target of the action of thapsigargin or downstream process of intracellular Ca 2+ signaling in mast cells

EFEK SENYAWA FLAVONOIDS DARI KEMUNING (Murraya paniculata [L.] Jack.) TERHADAP PELEPASAN HISTAMIN DARI KULTUR SEL MAST

Murraya paniculata [L.] Jack. (Kemuning) is a plant that grows widely in some areas of Indonesia. Studies related to this plant have been widely explored especially isolation of its active compounds. The plant contents several active compounds such as flanovoids. In the study, three flavonoid isolated from M. paniculata were evaluated for their effect on histamine release from mast cells (RBL-2H3 cells). These compounds were 3,3’,4’,5,5’,6,7,8-octamethoxyflavone; 3,3’,4’,5,5’,6,7-heptamethoxyflavone and 3, 3’, 4’, 5, 5’, 7–hexamethoxyflavone. The histamine inducers used in the study were DNP24-BSA dan thapsigargin, inducing the histamine release immunologically and non-immunologically, respectively. In the study, heptamethoxyflavone and hexamethoxyflavone did not influence the histamine release from mast cells significantly. However, octamethoxyflavone could increase the histamine release from RBL-2H3 cells in absence and presence the histamine inducers. The flavanoid could increase the release of histamine up to 50 %. Based on the results, polymethoxy moieties at the structure of flavonoid have a significant role to emerge the histamine-release stimulating effect from mast cells. Murraya paniculata [L.] Jack. merupakan tanaman yang tumbuh di Indonesia, dikenal dengan nama Kemuning. Penelitian mengenai tanaman ini telah banyak dilakukan, terutama isolasi senyawa aktifnya. Tanaman ini mempunyai kandungan senyawa aktif, diantaranya senyawa turunan flavonoid. Pada penelitian, tiga senyawa flavonoid yang diisolasi dari M. paniculata diuji aktivitasnya terhadap pelepasan histamin dari kultur sel mast yaitu sel RBL-2H3. Ketiga senyawa tersebut adalah 3,3’,4’,5,5’,7–heksametoksiflavon; 3,3’,4’,5,5’,6,7-heptametoksiflavon; dan 3,3’,4’,5,5’,6,7,8-oktametoksiflavon. Induktor pelepasan histamin yang digunakan adalah DNP24-BSA dan thapsigargin. Keduanya berturut-turut menginduksi secara imunologis dan non-imunologis. Hasil penelitian menunjukkan bahwa senyawa heptametoksiflavon dan heksametoksiflavon cenderung tidak mempengaruhi pelepasan histamin dari sel mast. Namun, oktametoksiflavon dapat meningkatkan pelepasan histamin dari sel mast baik tanpa induksi maupun terinduksi dengan DNP24-BSA atau thapsigargin. Senyawa tersebut mampun meningkatkan pelepasan histamin hingga 50%. Dari hasil tersebut, penambahan gugus polimetoksi pada struktur flavonoid berpotensi dapat menghasilkan efek pelepasan histamin dari sel mast. 

Effects of pentagamavunon-0 on histaminemediated hyperresponsive airway in asthmatic models : in-vitro in-vivo

Asthma  is  a  chronic  inflammatory  airway  disease  involving  reversible airway  constriction  and  airway  hyperresponsiveness  (AHR)  to  allergens,  airway edema,  and  increased  mucus  secretion  tumor  cells.  To  date,  exploration  of antiasthmatic  drug  is  still  being  studied  both  from natural  products  or  sinthetic processes.  One of the sinthetic compound is pentagamavunon-0  (PGV-0)  which possesses anti-inflammatory and inhibitory effects on histamine release from rat mast cells. The aim of the study is to look at the effects of PGV-0 on histaminemediated hyperresponsive airway in asthmatic models (in vitro and in vivo studies).  In  vitro  study  was  conducted  using  isolated  organ  technique  with isotonic  transducer.  The  results  have  shown  that  PGV-0  could  not  inhibit  the contraction  of  isolated  guinea  pig  trachea  induced  by  histamine.  PGV-0 did not change  the  pD2 and  Emax  values  of  histamine  on  trachea  smooth  muscle.  The finding  indicates  that  PGV-0  does  not  have  affinity and  intrinsic  activity  on  H-1 histaminergic  receptor  in  trachea  smooth  muscle.  In vivo  study,  we  sensitized the  rats  with  ovalbumin  (OVA)  to  develop  the  airway hyperreactivity  to histamine.  Histamine  level  in  bronchoalveolar  lavage  fluid  (BALF)  and  airway tissue  were  determined  using  HPLC-fluorometry.  Multiple  exposures  of ovalbumin  significantly  histamine  level  in  BALF  by  74.51±5.33  pmol/mL  or 6-times  higher  than  this  of  control  saline  group.  Oral  administration  of  PGV-0 (40  mg/kg  BW)  significantly  decreased  the  histamine accumulation  in  BALF  to 30  %  of  the  value  of  control  group  in  asthmatic  rats.  Besides,  PGV-0 significantly  prevented  the  histamine  decrease  in  asthmatic  rats  to  37.8  % trachea, and 34.2 % in bronchus. However, PGV-0 did not succeed to prevent the histamine decrease in the lung of asthmatic rats.  The result of the study may provide useful information for further discovering pharmacological synthetic compound for treatment of allergic inflammatory asthma.Key words: asthma, curcumin, pentamavunon-0, histamine, airway hyperresponsiveness