ANTIPROLIFERATIVE AND APOPTOTIC EFFECTS OF FICUS PSEUDOPALMA BLANCO (MORACEAE) AGAINST HEPATOCARCINOMA (HepG2)

Objectives.Hepatocellular carcinoma is the third most frequent cause of cancer-related death in the Philippines.  The prevalence of this disease continuously pose a great challenge to the medical and health sectors of the country, especially in discovering safe, efficacious and cheap alternative medicines with less or no adverse side effects.  Among our endemic medicinal plant species that belong to Moraceae family, Ficus pseudopalma remains understudied.  To our knowledge, there are no reports into the molecular mechanisms underlying the growth inhibitory effects of F.  pseudopalma against hepatocellular carcinoma.  We provided evidence that F.  pseudopalma enhances apoptotic cell death of HepG2, specifically targeting DNA damage. Methods. To assess the apoptotic and genotoxic abilities of the plant extract, trypan blue exclusion test, MTT, Comet, TUNEL and Caspase-3 activations assays were performed.  Results. Results showed that the crude extract of F. pseudopalma dose-dependently inhibited cell growth of HepG2 recording an IC50 of 353.342µg/mL.  Interestingly, all assays indicated apoptotic DNA fragmentation of HepG2 cells at concentrations of 300µg/ml and 1000µg/ml.  In particular, changes in morphological features such as aggregation of chromatin, cell shrinkage, and formation of apoptotic bodies were also observed in treated HepG2 cells.Conclusion. The data strongly suggest that F. pseudopalma may be used as an alternative, natural, cheap, readily available, and potent cancer chemoprotective agent by inhibiting cell growth and promoting cell death.  The study provides the first molecular evidence on the plant that induces apoptosis for the development of phytotherapeutic regimen for cancer prevention or treatment. Keywords:Ficus pseudopalma, cytotoxicity, apoptosis, Comet assay, TUNEL assay, caspase assa

Effect of Lactobacillus salivarius Dietary Supplementation on the Antioxidant Biomarkers of the Freshwater Shrimp Macrobrachium rosenbergii

The shrimp aquaculture is one of the most important livelihoods in the Philippines. Although there is an observed boost in the production of shrimps in the Philippines, there are remaining problems that are needed to be resolved. Bacterial and viral infections in shrimps have significantly affected its mortality, resulting in lower production and yield. In order to prevent or lessen this effect, various researches have been made to increase the resistance of shrimps against infections. In this study, a probiotic microorganism, Lactobacillus salivarius was used as a dietary supplementation for Macrobrachium rosenbergii and see its influence on the antioxidant biomarkers and temperature stress tolerance of the shrimp. Three concentrations of L. salivarius (1.0x107, 1.0x108, and 1.0x109 cells/g) were initially tested for its effect towards the oxidative stress tolerance and the oxygen consumption of the shrimp. The study involved the use of various antioxidant biomarkers including total glutathione (tGSH), catalase enzyme activity, and lipid peroxidation. Results showed that 1.0x108 cell/g of L. salivarius was the most effective concentration of probiotic microorganism per weight of feeds in improving the metabolic rate of the shrimp. Oxidative stress tolerance levels were observed to be increasing in terms of the catalase reaction (12.46 and 21.74 nmole/min) and lipid peroxidation (1.4X10-7 and 1.49X10-7 nmole MDA/mg protein) on both hepatopancreas and muscles tissues when the shrimp was fed with supplemented feeds. Also, high oxidant stress biomarkers were more evident on the hepatopancreas than the muscles

Lupeol: An antioxidant triterpene in Ficus pseudopalma Blanco (Moraceae)

Objective: To assess the antioxidant activity of Ficus pseudopalma Blanco (Moraceae) (F. pseudopalma) and characterize the active components present in it. Methods: Column chromatography of crude ethanol leaf extract of F. pseudopalma was performed and seven fractions were obtained, labeled as F1, F2, F3, F4, F5, F6, F7. DPPH, FRAP, Griess, Fenton and superoxide radical scavenging assays were performed to assess the antioxidant ability of the fractions. Thin layer chromatography (TLC), high performance liquid chromatography and Fourier transfer infrared spectroscopy (FTIR) were performed to identify and characterize the bioactive component present in each fractions of F. pseudopalma. Results: DPPH and FRAP assay showed that F5, F6 and F7 exhibited the good proton accepting ability and reducing power as compared to the other fractions. All fractions exhibited a good nitric oxide radical scavenging activity wherein F1, F2 and F3 showed the highest inhibition. However, all of the fractions exhibited a stimulatory activity on hydroxyl and superoxide radicals. Lupeol matched one of the spots on the thin layer chromatography chromatogram of the fractions. Linear gradient high performance liquid chromatography and spiking of lupeol with the fraction revealed the presence of 5.84 mg/L lupeol in F6. Infrared spectra of the fractions revealed the presence of C-C, OH, aromatic C=C and C=O groups. Conclusions: The identified lupeol in F. pseudopalma may be responsible for the exhibited antioxidant property of the plant. Furthermore, knowing the antioxidant capability of the plant, F. pseudopalma can be developed into products which can help prevent the occurrence of oxidative stress related diseases