This study investigated the effects of different percentages of ethanol (0 - 100%), extraction times (1 - 5 h) and temperatures (25 - 60°C) on total phenolic content (TPC) and antioxidant activity (AA) of sapodilla pulp and peel. TPC was determined by Folin-Ciocalteu reagent method, while AA was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay, ferric reducing antioxidant power (FRAP) assay and β-carotene bleaching (BCB) assay. Based on the optimal extraction conditions used, sapodilla pulp extract had TPC of 3.89 mg GAE/g, 63.20% of DPPH scavenging activity, 4.30% of ABTS scavenging activity, 19.17% of BCB activity, and FRAP value of 15.24 mg TE/g; while its peel extract had TPC of 9.23 mg GAE/g, 92.95% of DPPH scavenging activity, 5.36% of ABTS scavenging activity, 8.14% of BCB activity, and 27.85 mg TE/g (FRAP value). Using the optimal extraction conditions for sapodilla pulp (40% ethanol as extraction solvent that extracted at 60°C for 4 h) and sapodilla peel (80% ethanol and 2 h extraction time at 40°C), highest antioxidants can be extracted from the pulp and peel.
Manilkara zapota (L.) P. Royen (Family: Sapotaceae), commonly called as sapodilla, has been applied as traditional folk medicine for diarrhea and pulmonary infections. Conventional therapy in colorectal cancer is not likely effective due to undesirable outcomes. The anti-colon cancer properties of Manilkara zapota leaf water extract have yet to be investigated thus far. Therefore, our present study aimed to evaluate the ability to induce apoptosis and the underlying mechanisms of Manilkara zapota leaf water extract against human colorectal cancer (HT-29) cells. The cytotoxicity of Manilkara zapota leaf water extract was screened in different cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) analyses. The morphological changes in HT-29 cell lines after exposure to Manilkara zapota leaf water extract were viewed under fluorescence and inverted light microscope. The apoptotic cell was measured by Annexin V-propidium iodide staining. The caspase-3 and -8 activities were assessed by colorimetric assay. Overall analyses revealed that treatment with Manilkara zapota leaf water extract for 72 h can inhibit the viability of HT-29 cells. Incubation with Manilkara zapota leaf water extract for 24, 48, and 72 h significantly increased (p < 0.05) the total apoptotic cells compared to the control. Treatment with 21, 42, and 84 μg/mL of Manilkara zapota leaf water extract for 72 h triggered both caspase-3 and -8 activities in a concentration-dependent pattern. We also found that the catalase level in the two treatment groups (21 and 42 μg/mL) was significantly elevated after 24 h incubation. Incubation with Manilkara zapota leaf water extract for 72 h triggered the transcriptional elevation of the adenomatous polyposis coli (APC), glycogen synthase kinase 3β (GSK3β), AXIN1, and casein kinase 1 (CK1). The β-catenin mRNA levels were reduced accordingly when the concentration of the Manilkara zapota leaf water extract was increased. Our results suggested that Manilkara zapota leaf water extract offer great potential against colorectal cancer through modulation of Wnt/β-catenin signaling pathway, caspase-dependent pathway, and antioxidant enzyme.
Introduction: Manilkara zapota (L.) P. Royen or sapodilla is a fruit-bearing tree that has been cultivated mainly in tropical areas including Mexico and South East Asia. The fruits and the other parts of M. zapota plant have been used since ages ago for various medicinal purposes. However, the data on the antioxidant properties of various parts of M. zapota is limited. Therefore, we aimed to measure the content and capacity of antioxidants in various M. zapota plant parts and also to screen the phytoconstituents present in the part with the highest antioxidant content and capacity. Methods: The in vitro antioxidant evaluation including the content of total phenolic (TPC) and total flavonoids (TFC) as well as β-carotene bleaching and 1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability of the leaves, seeds, flesh, and peels of M. zapota extracted by aqueous and ethanol were determined. The plant part that exhibited the highest TPC, TFC, and antioxidant capacity was selected for phytoconstituents identification using liq- uid chromatography mass spectrometry. Results: M. zapota leaves aqueous extract exhibited the highest TPC, TFC, and antioxidant capacities and therefore selected for phytoconstituents identification. Our study provide additional data in which a total of 39 phytoconstituents have been identified in the M. zapota leaves including m-coumaric acid, quinic acid, robinetinidol-4alpha-ol, isoorientin 6’’-O-caffeate, apocynin A, and C16 Sphinganine. Conclusion: Thus, our study revealed that M. zapota leaves aqueous extract has potential as a promising naturally-occurring an- tioxidant candidate which could be useful for medicinal and nutritional functions.
Malaysia has a rich diversity of medicinal plants and some of them inhibit xanthine oxidase (XO), which can be introduced as new natural sources of gout medication and a substitute for synthetic xanthine oxidase inhibitors (XOI). The degree of XO inhibitory activity was determined by measuring the absorbance spectrophotometrically at 295 nm, which is associated with uric acid formation. Our preliminary screening study had employed the use of distilled water, 70% methanol and absolute ethanol to extract XOI from twenty parts of five plant species, namely, Averrhoa carambola, Carica papaya, Dimocarpus longan malesianus, Manilkara zapota and Salacca zalacca. These plants were selected based on their frequent medicinal usages by local folks. The results have shown that an aqueous extract of Carica papaya mature leaves has promising activity to inhibit XO up to 75.68 ± 0.1%. Statistical experimental design were employed to optimize the selected sample (dried Carica papaya leaves: distilled water) on extraction of XOI and the maximum XOI percentage of 86.93 ± 1.9% was obtained, which exhibited only 6.76% less than the activity exhibited by allopurinol (93.69 ± 0.2%), a commercial XOI. The comparison was made between allopurinol and optimized extract on the basis of IC50concentrations. Allopurinol showed IC50 value of 3.74 μg/ml that is considerably lower as compared to the optimized sample (4.33 μg/ml).
Manilkara zapota (L.) P. Royen (family: Sapotaceae) is commonly called sapodilla, or locally known as ciku. The detailed mechanisms underlying Manilkara zapota leaf methanol extract against HeLa human cervical cancer cells have yet to be investigated. Therefore, our present study is designed to investigate the ability to induce apoptosis and the underlying mechanisms of Manilkara zapota leaf methanol extract inducing cytotoxicity in HeLa cells. The apoptotic cell death was assessed using Annexin V-propidium iodide staining. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential activities were measured using dichlorodihydrofluorescein diacetate and MitoLite Orange, respectively, by NovoCyte Flow Cytometer. Bax and Bcl-2 expression were evaluated using Enzyme-Linked Immunosorbent Assay. Caspase-3 activity was determined using a colorimetric assay. The associated biological interaction pathways were evaluated using quantitative real-time PCR. Our data showed that HeLa cells were relatively more sensitive to Manilkara zapota leaf methanol extract than other cancer cell lines studied. Overall analyses revealed that Manilkara zapota leaf methanol extract can inhibit the viability of HeLa cells, induce mitochondrial ROS generation, and inhibit nuclear factor-kappa B (NF-κB) and epidermal growth factor receptor (EGFR) transcriptional activities. Our results suggested that Manilkara zapota leaf methanol extract might represent a potential anticervical cancer agent.
Manilkara zapota (L.) P. Royen, called sapodilla, or locally known as ciku, belongs to the family Sapotaceae. We found that Manilkara zapota leaf water extract has cytotoxic effect against human hepatocellular carcinoma (HepG2) cell line in our earlier study. Therefore, this study aimed to explore the anticancer properties of Manilkara zapota leaf water extract in HepG2 cells. We also aimed to unravel yet undiscovered mechanisms and identified several expressed genes whose functions in cytotoxicity activity of Manilkara zapota leaf water extract in HepG2 cells have not been well-studied. The apoptosis and intracellular reactive oxygen species (ROS) activities were analyzed using Annexin V-propidium iodide staining and dichlorodihydrofluorescein diacetate, respectively, by NovoCyte Flow Cytometer. Bax and Bcl-2 expression were assessed using Enzyme-Linked Immunosorbent Assay. The associated molecular pathways were evaluated by quantitative real-time PCR. Overall analyses revealed that Manilkara zapota leaf water extract can increase percentage of early apoptotic cells, induce the formation of ROS, upregulate c-Jun N-terminal kinase 1 (JNK1) and inducible nitric oxide synthase (iNOS), and reduce Akt1 and vascular endothelial growth factor A (VEGFA) transcriptional activities. Our data suggest that Manilkara zapota leaf water extract can suppress the growth of HepG2 cells via modulation of ERK1/2/Akt1/JNK1 transcriptional expression.