Antiproliferation activity of God's crown fruit (Phaleria macrocarpa) extract and fractions against MCM-B2 breast cancer cells

Breast cancer is a disease caused by abnormal cell proliferation in the breast. God’s crown fruit (Phaleria macrocarpa)and its seed have potential as an antiproliferation of cancer cells. It contains active compounds such as flavonoids,alkaloids, polyphenols, and tannins. The sample of God’s crown fruit was obtained by extraction and fractionationusing the maceration method. Cytotoxicity of extracts and fractions was determined using Brine Shrimp Lethality Testmethod. Antiproliferation activity test of God’s crown fruit against MCM-B2 was performed using the hemacytometermethod. The God’s crown fruit sample consists of crude ethanol extract, n-hexane fraction, ethyl acetate fraction, andwater fraction. Lethal concentration 50 (LC50) values in crude ethanol extract, n-hexane fraction, ethyl acetate fraction,and water fraction were 13.72, 147.55, 405.81, and 149.55 ppm, respectively. The concentration of the test sample wasdirectly used for the antiproliferation activity test on MCM-B2 cells. God’s crown fruit can act as antiproliferation ofMCM-B2. The smallest concentration of those samples has inhibited MCM-B2 cell proliferation which is 3.5 ppmcrude ethanol extract lower than 100 ppm doxorubicin. The maximum percentage of the antiproliferation activity ofcrude ethanol extract (56 ppm) was able to inhibit MCM-B2 cell proliferation by 58.28% while doxorubicin (100 ppm)by 31.2%. This is due to the fact that crude ethanol extract has a lot of complex polar phytochemical content. The crudeethanol extract compounds inhibit MCM-B2 cell proliferation synergistically

Isolation and identification of compounds from Phaleria macrocarpa (Scheff.) Boerl fruit extract

Objective:To identify and isolate the chemical compounds of Phaleria macrocarpa (P.macrocarpa) fruit ethanolic extract.Methods:Dried fruit of P.macrocarpa was extracted with 90％ ethanol and partitioned between n-hexane/H2O and ethyl acetate/H2O.The organic layer was fractionated by various stationary phase and identified by using combined data of ID [(proton nuclear magnetic resonance (NMR),carbon-13 NMR)],2D-NMR (heteronuclear multiplequantum correlation and heteronuclear multiple-bond correlation),and mass spectrum.Results:Purification of n-hexane and ethyl acetate fractions from ethanolic extract of P.macrocarpa fruit resulted in isolation of nine compounds.Conclusions:A new compound was isolated and identified as glyceryl pentacosanoate.Also,two xanthones,which are 1,7-dihydroxy-3,6-dimethoxyxanthone and 1,6,7-trihydroxy-3-methoxyxanthone,are firstly reported to be isolated from P.macrocarpa.

Inhibition of Mahkota Dewa (Phaleria macrocarpa) bioactive fraction on proliferation of human retinoblastoma tumor cells Y-79 through suppression of mRNA level of cyclin E

Objecive:To prove the molecular mechanisms of Mahkota Dewa (Phaleria macrocarpa) in suppressing proliferation of human retinoblastoma cells through suppression of cell cycle's gene-rcgulators expression.Methods:In this study,the molecular mechanism of anti-tumor effect of fractioned extract of Phaleria macrocarpa (DLBS1425) in human retinoblastoma cells Y-79 was investigated by measuring the tumor cells viability,the assessment of population profiles of tumor cells in the cell cycle,and the mRNA concentration of pi6,p21,p53,cyclin D,cyclin E,and E2F.Results:DLBS1425 showed an inhibition effects towards proliferation of Y-79 cell line.Inhibition of proliferation was shown by suppression of cell cycle progression.DLBS1425 downregulated cyclin E,a G1 phase regulator gene of cell cycle,in dosedependent manner without affecting p53-p21 pathway.In the other word,DLBS1425 inhibits cell proliferation through suppression of cyclin E independently towards conventional proliferation pathway.Conclusions:Our results suggest that DLBS 1425 is a potential anticancer agent which targets genes involved in cell proliferation in human retinoblastoma cells which make it pharmacologically ideal for the prevention and/or treatment of retinoblastoma cancer.

Isolation and identification of compounds from Phaleria macrocarpa (Scheff.) Boerl fruit extract

Objective To identify and isolate the chemical compounds of Phaleria macrocarpa (P. macrocarpa) fruit ethanolic extract. Methods Dried fruit of P. macrocarpa was extracted with 90% ethanol and partitioned between n-hexane/H

Inhibition of Mahkota Dewa (Phaleria macrocarpa) bioactive fraction on proliferation of human retinoblastoma tumor cells Y-79 through suppression of mRNA level of cyclin E

Objective To prove the molecular mechanisms of Mahkota Dewa (Phaleria macrocarpa) in suppressing proliferation of human retinoblastoma cells through suppression of cell cycle's gene-regulators expression. Methods In this study, the molecular mechanism of anti-tumor effect of fractioned extract of Phaleria macrocarpa (DLBS1425) in human retinoblastoma cells Y-79 was investigated by measuring the tumor cells viability, the assessment of population profiles of tumor cells in the cell cycle, and the mRNA concentration of p16, p21, p53, cyclin D, cyclin E, and E2F. Results DLBS1425 showed an inhibition effects towards proliferation of Y-79 cell line. Inhibition of proliferation was shown by suppression of cell cycle progression. DLBS1425 downregulated cyclin E, a G1 phase regulator gene of cell cycle, in dose-dependent manner without affecting p53–p21 pathway. In the other word, DLBS1425 inhibits cell proliferation through suppression of cyclin E independently towards conventional proliferation pathway. Conclusions Our results suggest that DLBS1425 is a potential anticancer agent which targets genes involved in cell proliferation in human retinoblastoma cells which make it pharmacologically ideal for the prevention and/or treatment of retinoblastoma cancer.

Phaleria macrocarpa Fruits Methanolic Extract Reduces Blood Pressure and Blood Glucose in Spontaneous Hypertensive Rats (SHR).

Methanolic extract of Phaleria macrocarpa fruits was evaluated for the anti-hypertensive and anti-hyperglycaemic capacity. For a period of three weeks, 18 male young Spontaneous Hypertensive Rats (SHR) were divided into three experimental groups and received commercial rat pellets as their maintenance diet. At the same duration, SHR in Group 1 received 2 ml/kg/day of distilled water and served as a negative control. Meanwhile, SHR in Group 2 and Group 3 received 0.5 mg/kg/day of Telmisartan (positive control) and 500 mg/kg/day P. macrocarpa fruits methanolic extract, respectively. Bodyweight, arterial blood pressure and blood glucose were measured on a weekly basis. All SHR showed significant increased (p<0.05) in their bodyweight but did not differ significantly (p>0.05) between groups. Blood pressure and blood glucose significantly decreased (p<0.05) in SHR treated with Telmisartan and P. macrocarpa compared to negative control. These results lead us to conclude that P. macrocarpa fruits methanolic extract exhibit anti-hypertensive and anti-hyperglycaemic activities.

Production of secondary metabolite E2.2 from Phaleria macrocarpa endophytic fungus

Objective: To isolate new endophytic fungus from Phaleria macrocarpa (P. macrocarpa) that is able to produce E2.2 compound. Methods: Endophytic fungi were isolated from P. macrocarpa. Morphological and molecular identification was done to determine the species of the endophytic fungus. High performance liquid chromatography was used to determine the ability of this fungus to produce E2.2 compound and to quantify the total yield of E2.2 from fungal fermen-tation. Fermentation process was optimized by observing suitable medium, pH and length of fermentation process. Phloroglucinol and gallic acid addition were examined to determine the effect of each compound on E2.2 production. Results: One endophytic fungus was successfully isolated from P. macrocarpa plant. Morphological and molecular identification showed that it was a Colletotrichum gloeo-sporioides which belonged to Glomerellaceae family. This fungus showed highest pro-duction of E2.2 when incubated in potato dextrose broth with initial pH value of the medium at 5, and was incubated for 15 days. Phloroglucinol was found to better enhance E2.2 production. Conclusions: Colletotrichum gloeosporioides found in P. macrocarpa plant is prom-ising as a potential alternative source of E2.2.

Standardized bioactive fraction of Phaleria macrocarpa (Proliverenol) prevents ethanol-induced hepatotoxicity via down-regulation of NF-kB-TNFα-caspase-8 pathway

Objective: To verify that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. Methods: Activity of Proliverenol against ethanol-induced apoptosis was evaluated at mRNA and protein levels in HepG2 cell exposed to Proliverenol for 1 and 3 h. Results: Proliverenol conferred hepatoprotective activity through increasing cell survival up to 53%-69% via up-regulation of APEX1 DNA repair enzyme for 3.0-4.7 fold and down-regulating of nuclear factor-kB, tumor necrosis factorα and caspase-8 expression, allowing them to prevent 4.5-6.9 fold of alanine aminotransferase (ALT) leakage in HepG2 cells. Our finding revealed that Proliverenol repressed expression of ALT, which is significantly important as possible alternative mechanism for increased blood transaminase activities. In addition, the result also showed that caspase-8 pathway seemed to be involved in the molecular pathway rather than directly inducing mitochondrial damage. Conclusions: The data support our hypothesis that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. We propose that Proliverenol pro-vides hepatoprotective activity through up-regulating expression of APEX1 that repress DNA fragmentation, and down-regulating expression of nuclear factor-kB, tumor ne-crosis factorαand caspase-8, which therefore repress ALT leakage and its expression.

Standardized bioactive fraction of Phaleria macrocarpa (Proliverenol) prevents ethanol-induced hepatotoxicity via down-regulation of NF-κB-TNFα-caspase-8 pathway

Objective To verify that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. Methods Activity of Proliverenol against ethanol-induced apoptosis was evaluated at mRNA and protein levels in HepG2 cell exposed to Proliverenol for 1 and 3 h. Results Proliverenol conferred hepatoprotective activity through increasing cell survival up to 53%–69% via up-regulation of APEX1 DNA repair enzyme for 3.0–4.7 fold and down-regulating of nuclear factor-κB, tumor necrosis factorα and caspase-8 expression, allowing them to prevent 4.5–6.9 fold of alanine aminotransferase (ALT) leakage in HepG2 cells. Our finding revealed that Proliverenol repressed expression of ALT, which is significantly important as possible alternative mechanism for increased blood transaminase activities. In addition, the result also showed that caspase-8 pathway seemed to be involved in the molecular pathway rather than directly inducing mitochondrial damage. Conclusions The data support our hypothesis that Proliverenol has a potential ability in protecting cells from ethanol-induced hepatotoxicity. We propose that Proliverenol provides hepatoprotective activity through up-regulating expression of APEX1 that repress DNA fragmentation, and down-regulating expression of nuclear factor-κB, tumor necrosis factorα and caspase-8, which therefore repress ALT leakage and its expression.

Production of secondary metabolite E2.2 from Phaleria macrocarpa endophytic fungus

Objective To isolate new endophytic fungus from Phaleria macrocarpa (P. macrocarpa) that is able to produce E2.2 compound. Methods Endophytic fungi were isolated from P. macrocarpa. Morphological and molecular identification was done to determine the species of the endophytic fungus. High performance liquid chromatography was used to determine the ability of this fungus to produce E2.2 compound and to quantify the total yield of E2.2 from fungal fermentation. Fermentation process was optimized by observing suitable medium, pH and length of fermentation process. Phloroglucinol and gallic acid addition were examined to determine the effect of each compound on E2.2 production. Results One endophytic fungus was successfully isolated from P. macrocarpa plant. Morphological and molecular identification showed that it was a Colletotrichum gloeosporioides which belonged to Glomerellaceae family. This fungus showed highest production of E2.2 when incubated in potato dextrose broth with initial pH value of the medium at 5, and was incubated for 15 days. Phloroglucinol was found to better enhance E2.2 production. Conclusions Colletotrichum gloeosporioides found in P. macrocarpa plant is promising as a potential alternative source of E2.2.