Metabolomic study and in silico approach of DLBS1442 as progesterone receptor agonist

Endometriosis prevalence has been known to be quite high among women of reproductive age and with pelvic painand/or infertility. The reason is that the estrogen level in the eutopic endometrium of women with endometriosis ishigher than in normal endometrium which may possibly be caused by the lack of interaction between progesteroneand progesterone receptor (PR). Dexa Laboratories of Biomolecular Sciences (DLBS) has developed DLBS1442,a bioactive fraction from Phaleria macrocarpa (Scheff) Boerl fruit, which has been found to be potential to treatsymptoms of primary dysmenorrhea and alleviate endometriosis. Therefore, the identification of DLBS1442 activecompounds which act as a PR agonist was necessary. Identification was performed using metabolomics study whichresulted in 14 compounds. Crystal structure of the PR with asoprisnil as the reference was obtained from PDB (4A2J).Virtual screening validation process was performed using Protein-Ligand ANT System (PLANTS) and Pythonbased Protein-Ligand Interaction Fingerprinting (PyPLIF). According to the virtual screening protocol validation,the highest Enrichment Factor (EF) 1% value was obtained with hydrogen interaction with GLN725 and ARG766residue. Virtual screening of the DLBS1442 metabolomics study showed that only glyceryl pentacosanoate exhibited alower Chem Piecewise Linear Potential (ChemPLP) than the cutoff. This compound might have a role as a PR agonistwhich supported the previous findings of DLBS1442 to alleviate endometriosis. However, this finding requires furtherin vitro and/or in vivo study to ensure the agonist activity of glyceryl pentacosanoate as a DLBS1442 active compound.

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.

Trombinol, a bioactive fraction of Psidium guajava, stimulates thrombopoietin expression in HepG2 cells

Objective To study the regulation of trombinol on thrombopoietin, an essential regulator of thrombocyte production. Methods Effect of trombinol on thrombopoietin regulation was evaluated at the mRNA and protein levels in human hepatoma HepG2 cells. The mRNA expressions were revealed by PCR and real-time PCR, while the protein expressions were analyzed using western blotting and human ELISA kit. Statistical differences between the test were determined by student's t-test with P < 0.05 was considered statistically significant. Results Trombinol significantly increased the expression of thrombopoietin at the level of mRNA and protein secretion in HepG2 cell lines. Trombinol with the concentration of 15 μg/mL, positively induces 2.5-fold of thrombopoietin expression. Up-regulation of GABP, a transcription factor of thrombopoietin, is suggested to be involved in cellular regulatory mechanisms of trombinol. Here, our result shows convincing evidence that trombinol affects the thrombopoietin productions in vitro. This molecular explanation of thrombopoietin's stimulating function is in line with the traditional use of Psidium guajava for treatment of diseases involving thrombocytopenia. Conclusions Thrombopoietin stimulating function of trombinol could be potentially considered as one of alternative treatment for thrombocytopenia-related cases, including post chemotherapy shock, dengue fever and liver failure.

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.

Potential effect of striatin (DLBS0333), a bioactive protein fraction isolated from Channa striata for wound treatment

Objective To characterize proteins and other nutrients in striatin (DLBS0333), a bioactive protein fraction isolated from snakehead fish (Channa striata) and to investigate its wound healing activity. Methods Proteins and other constituents in striatin were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two dimension electrophoresis, immunoblotting assay, spectroscopy and high performance liquid chromatography. The wound healing activity of striatin was studied in vitro using 3T3 fibroblast cells and in vivo using wound-induced animal model. Various parameters related to wound healing process were evaluated. Results Striatin contained four major bioactive proteins with approximate molecular weight of 8.3, 10.9, 15.4 and 16.7 kDa. In addition to proteins, striatin also contained amino acids (10 essential and 7 non essential amino acids), fatty acids (palmitic acid, oleic acid, stearic acid, linoleic acid, arachidonic acid), vitamins (vitamin A, vitamin B