Ficus Deltoidea Var Research Articles

Effects of Standardized Aqueous Ethanolic Extract of Leaves of Ficus Deltoidea var. Trengganuensis on Blood Pressure in Spontaneously Hypertensive Rats

Background: Laboratory evidence pertaining to the antihypertensive activity of a number of Ficus deltoidea (FD) varieties remains undetermined, although extracts of some of these varieties have been shown to have angiotensin-converting enzyme (ACE) inhibitory activity in vitro. Objective: This study, therefore, evaluated the effect of a standardized aqueous-ethanolic extract of leaves of Ficus deltoidea var. trengganuensis (FDT) on blood pressure in spontaneously hypertensive rats (SHR). Methods: SHR were given either to 800, 1000 or 1200 mg kg-1 body weight of standardized aqueous- ethanolic extract of FDT or 10 mg kg-1 body weight of losartan or 0.5 ml of distilled water daily for four weeks. Results: SBP decreased significantly in FDT- and losartan-treated rats (p < 0.05). Components of RAAS and other serum and urinary parameters were not different between the groups except for Endothelin-1, which was significantly lower than that in the controls. Urinary calcium excretion was significantly higher in FDT-treated rats (p < 0.05). Conclusion: It seems that daily oral administration of FDT significantly lowers blood pressure in SHR through mechanisms that do not involve RAAS but may involve the endothelium or other yetto- be-determined mechanisms.

Ficus deltoidea Leaf Alters Oxidative Stress, Protein Homeostasis and Ubiquitin-Proteasome Pathways in Fatty Acid-Induced Cell Line.

Ficus deltoidea (mistletoe fig) is a shrub well known among locals in Malaysia primarily for its treatment of toothaches, colds and wounds. The aim of this study is to determine the potential of leaves, sourced from three different varieties of F. deltoidea, to exhibit antioxidant activity, a reduction of lipid concentration, and protein expression in steatosis-induced liver cell lines. The leaves of three F. deltoidea varieties, namely Ficus deltoidea var. angustifolia, Ficus deltoidea var. trengganuensis and Ficus deltoidea var. kunstleri, were subjected to water extraction. The resulting crude extracts were fractionated using water and ethyl acetate. Palmitic acid was used to induce lipid accumulation (steatosis) in human liver (WRL68) cells, before all the samples were tested for their lipid-reducing activity. Several proteomic approaches were incorporated. The changes in protein expression were determined using 2-dimensional gel electrophoresis separation, whereas identification of our protein spots of interest was carried out via matrix-assisted laser desorption/ionization time-of-flight. Ficus deltoidea var. kunstleri alone demonstrated the ability to reduce lipids at the highest tested concentration (200 µg/mL) and was, therefore, used for subsequent experiments. Treatment with Ficus deltoidea var. kunstleri was found to restore redox status by increasing superoxide dismutase and glutathione peroxidase amounts and decreasing malondialdehyde formation. Six proteins were successfully identified; these were heat shock protein beta-1 (HSPB1), proteasome subunit alpha type 1 (PSMA1), glutathione S-transferase omega 1 (GSTO1), peroxiredoxin-1 (PRDX1), histone H2B (HIST1H2BD) and ubiquitin c-terminal hydrolase L3 (UCHL3). Through bioinformatics analysis, it was found that these proteins were significantly involved in specific pathways such as oxidative stress (PRDX1 and GSTO1), protein homeostasis (HSPB1) and degradation (UCHL3 and PSMA1). F. deltoidea pretreatment was shown to reduce lipid accumulation, thus improving the redox status and protein homeostasis. This suggests the role of F. deltoidea as a preventive mechanism in non-alcohol fatty liver disease.

Blood pressure and urine metabolite changes in spontaneously hypertensive rats treated with leaf extract of Ficus deltoidea var angustifolia

Ficus deltoidea var angustifolia (FD-A) reduces blood pressure in spontaneously hypertensive rats (SHR) but the mechanism remains unknown. Changes in urine metabolites following FD-A treatment in SHR were, therefore, examined to identify the mechanism of its antihypertensive action. Male SHR were given either FD-A (1000 mg kg−1 day−1) or losartan (10 mg kg−1 day−1) or 0.5 mL of ethanolic-water (control) daily for 4 weeks. Systolic blood pressure (SBP) was measured every week and urine spectra data acquisition, on urine collected after four weeks of treatment, were compared using multivariate data analysis. SBP in FD-A and losartan treated rats was significantly lower than that in the controls after four weeks of treatment. Urine spectra analysis revealed 24 potential biomarkers with variable importance projections (VIP) above 0.5. These included creatine, hippurate, benzoate, trimethylamine N-oxide, taurine, dimethylamine, homocysteine, allantoin, methylamine, n-phenylacetylglycine, guanidinoacetate, creatinine, lactate, glucarate, kynurenine, ethanolamine, betaine, 3-hydroxybutyrate, glycine, lysine, glutamine, 2-hydroxyphenylacetate, 3-indoxylsulfate and sarcosine. From the profile of these metabolites, it seems that FD-A affects urinary levels of metabolites like taurine, hypotaurine, glycine, serine, threonine, alanine, aspartate and glutamine. Alterations in these and the pathways involved in their metabolism might underlie the molecular mechanism of its antihypertensive action.

The use of pattern recognition for classification of Indonesian ginger (Zingiber officinale var. amarum) based on antioxidant activities and FTIR spectra

Discriminating Ficus deltoidea var. bornensis from Different Localities by HPTLC and FTIR Fingerprinting Azierah Azemin, Saravanan Dharmaraj, Muhammad Razak Hamdan, Nashriyah Mat, Zhari Ismail4 and Khamsah Suryati Mohd

Ficus deltoidea ameliorates biochemical, hormonal, and histomorphometric changes in letrozole-induced polycystic ovarian syndrome rats

BackgroundInsulin resistance and hormonal imbalances are key features in the pathophysiology of polycystic ovarian syndrome (PCOS). We have previously shown that Ficus deltoidea var. deltoidea Jack (Moraceae) can improve insulin sensitivity and hormonal profile in PCOS female rats. However, biological characteristics underpinning the therapeutic effects of F. deltoidea for treating PCOS remain to be clarified. This study aims to investigate the biochemical, hormonal, and histomorphometric changes in letrozole (LTZ)-induced PCOS female rats following treatment with F. deltoidea.MethodsPCOS was induced in rats except for normal control by administering LTZ at 1 mg/kg/day for 21 days. Methanolic extract of F. deltoidea leaf was then orally administered to the PCOS rats at the dose of 250, 500, or 1000 mg/kg/day, respectively for 15 consecutive days. Lipid profile was measured enzymatically in serum. The circulating concentrations of reproductive hormone and antioxidant enzymes were determined by ELISA assays. Ovarian and uterus histomorphometric changes were further observed by hematoxylin and eosin (H&E) staining.ResultsThe results showed that treatment with F. deltoidea at the dose of 500 and 1000 mg/kg/day reduced insulin resistance, obesity indices, total cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL), malondialdehyde (MDA), testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) to near-normal levels in PCOS rats. The levels of high-density lipoprotein cholesterol (HDL), estrogen, and superoxide dismutase (SOD) are also similar to those observed in normal control rats. Histomorphometric measurements confirmed that F. deltoidea increased the corpus luteum number and the endometrial thickness.ConclusionsF. deltoidea can reverse PCOS symptoms in female rats by improving insulin sensitivity, antioxidant activities, hormonal imbalance, and histological changes. These findings suggest the potential use of F. deltoidea as an adjuvant agent in the treatment program of PCOS.

Effects of aqueous ethanolic ficus deltoidea var. kunstleri extract on lipid profile and inflammation in high cholesterol diet-induced atherosclerotic rabbits

Background and Aims: Ficus deltoidea var. kunstleri (FDK), a natural product has been reported to exhibit atheroprotective properties in vitro. However, its in vivo atheroprotective effects are unknown. This study aims to investigate the atheroprotective effects of aqueous ethanolic extract of FDK on high cholesterol diet (HCD)-induced early and established atherosclerosis in rabbits.

Antimutagenic, Cytoprotective and Antioxidant Properties of Ficus deltoidea Aqueous Extract In Vitro.

Ficus deltoidea var. deltoidea is used as traditional medicine for diabetes, inflammation, and nociception. However, the antimutagenic potential and cytoprotective effects of this plant remain unknown. In this study, the mutagenic and antimutagenic activities of F. deltoidea aqueous extract (FDD) on both Salmonella typhimurium TA 98 and TA 100 strains were assessed using Salmonella mutagenicity assay (Ames test). Then, the cytoprotective potential of FDD on menadione-induced oxidative stress was determined in a V79 mouse lung fibroblast cell line. The ferric-reducing antioxidant power (FRAP) assay was conducted to evaluate FDD antioxidant capacity. Results showed that FDD (up to 50 mg/mL) did not exhibit a mutagenic effect on either TA 98 or TA 100 strains. Notably, FDD decreased the revertant colony count induced by 2-aminoanthracene in both strains in the presence of metabolic activation (p < 0.05). Additionally, pretreatment of FDD (50 and 100 µg/mL) demonstrated remarkable protection against menadione-induced oxidative stress in V79 cells significantly by decreasing superoxide anion level (p < 0.05). FDD at all concentrations tested (12.5–100 µg/mL) exhibited antioxidant power, suggesting the cytoprotective effect of FDD could be partly attributed to its antioxidant properties. This report highlights that F. deltoidea may provide a chemopreventive effect on mutagenic and oxidative stress inducers.

Phytochemical and In Vitro Genotoxicity Studies of Standardized Ficus deltoidea var. kunstleri Aqueous Extract

The present study was carried out to assess the genotoxicity potential of Ficus deltoidea var. kunstleri aqueous extract (FDAE) using standard in vitro assays. The DNA damage of V79B cells was measured using the alkaline comet assay treated at 0.1 mg/mL (IC10) and 0.3 mg/mL (IC25) of FDAE together with positive and negative controls. For in vitro micronucleus assay, the V79B cells were treated with FDAE at five different concentrations (5, 2.5, 1.25, 0.625, and 0.3125 mg/mL) with and without S9 mixture. The bacteria reverse mutation assay of FDAE was performed on Salmonella typhimurium strains TA98, 100, 1535, 1537, and Escherichia coli strain WP2uvrA using pre-incubation method in the presence or in the absence of an extrinsic metabolic system (S9 mixture). FDAE at 0.1 and 0.3 mg/mL significantly increased DNA damage in both comet tail and tail moment (p < 0.05). No significant changes were detected in the number of micronucleated cell when compared to control. Tested at the doses up to 5000 µg/plate, the FDAE did not increase the number of revertant colonies for all strains. In conclusion, further investigation needs to be conducted in animal model to confirm the non-genotoxicity activities of FDAE.

Effect of Hydrogen Peroxide on Plant Growth, Photosynthesis, Leaf Histology and Rubisco Gene Expression of the Ficus deltoidea Jack Var. deltoidea Jack

The present study was carried out to investigate the effects of hydrogen peroxide (H2O2) on the growth, photosynthesis, leaf histology, mineral uptake, phytochemical accumulation, and Rubisco gene expression in Ficus deltoidea var. deltoidea, a slow growing medicinal plant. The F. deltoidea plants were spray-treated with 0 (control), 8, 16, 30 and 60 mM H2O2 under field conditions. The results showed that the applications of 30 mM H2O2 increased the plant height, leaf number, and syconium number by 26%, 9% and 2275%, respectively, over the control specimen. Photosynthetic rates, transpiration, stomatal conductance and chlorophyll content of F. deltoidae plants increased significantly after treatment of 30 mM H2O2. A lower concentration (8 and 16 mM) of H2O2 also improved the leaf histological properties such as tracheary elements and other vascular cells. In addition, treatments with 30 mM H2O2 increased calcium, potassium, total phenolic and flavonoids content, in F. deltoidea plants, by 14%, 15%, 12% and 72%, respectively. In addition, H2O2 treatment at 30 mM concentration downregulated the Rubisco gene with a between 1 and 5 times fold gene expression change. From this study, it can be concluded that spraying with 30 and 16 mM of H2O2 once a week enhanced growth, photosynthesis, mineral accumulation, phenol and flavonoid content, as well as aided Rubisco gene expression, under field conditions.

Potential applications of enzymes on the extraction of vitexin from dried Mas Cotek leaves

Mas Cotek or Ficus deltoidea var. deltoidea leaves extracts contain various beneficial properties such as antidiabetic, antihypoglycemic and antioxidant activities, which can be enzymatically extracted in water. Vitexin is one of the antioxidants and remarkable compound of flavone C-glycosides found in Mas Cotek leaves. Cellulase is one of the essential enzymes applied to hydrolyze cellulose into fermentable sugars and therefore, this enzyme was used to extract vitexin in this study. Single parameter approach was used to determine factors influencing the enzymatic-mediated extraction. These factors were enzyme concentration, sample-to-water ratio, temperature, incubation time and agitation. The maximum yield of vitexin found was 0.547% under the optimum conditions: sample-to-water ratio 1:10 g/mL, enzyme concentration 0.4%, incubation time of 240 min, temperature of 50 ± 0.02°C and stirring speed of 200 rpm. The highest amount of reducing sugar obtained was 4.039 mg/mL with the use of 0.4% cellulose. A correlation was also observed between the yield of vitexin and reducing sugars. Key words: Cellulose, enzymatic assisted extraction, vitexin, reducing sugars, cellulose, Mas Cotek, Ficus deltoidea var. deltoidea, antioxidants.

The Effects of Hydrogen Peroxide on Plant Growth, Mineral Accumulation, as Well as Biological and Chemical Properties of Ficus deltoidea

Hydrogen peroxide (H2O2) is defined as a reactive oxygen species (ROS), able to cause damage to a variety of cellular structures. On the other hand, recent work has demonstrated that H2O2 can also act as a potent signaling molecule that mediates various physiological and biochemical processes in plants. This study was carried out to investigate the effects of H2O2 on the growth, mineral nutrient accumulation, as well as the biologic and chemical properties of Ficus deltoidea var. deltoidea. F. deltoidea plants were spray-treated with 0- (control), 8-, 16-, 30- and 60-mM H2O2 under field conditions. Plant height, leaf area, chlorophyll content, net photosynthetic rate, stomatal conductance and quantum yield of the F. deltoidea plants significantly increased after treatment with 16 and 30-mM H2O2. The results indicate that 60-mM H2O2 increased the accumulation of arsenic, iron and sodium content in the leaves of F. deltoidea. On the other hand, 8-mM H2O2 significantly enhanced the accumulation of arsenic, iron, calcium and potassium content in the syconium of F. deltoidea plants. In addition, H2O2 treatment did not produce any significant effects on antimony and magnesium accumulation in the leaves or the syconium of F. deltoidea plants. The results show that the F. deltoidea plant has strong antidiabetic properties and its α-glucosidase activity increased in treated plants compared to standard acarbose. Hydrogen peroxide, particularly in concentrations of 16 and 30 mM, increased the antioxidant activity, total phenolic and flavonoid content and the vitexin and isovitexin content. There was a positive correlation between antioxidant activity with total phenol and total flavonoid content in H2O2-treated plants. The quantitative analysis by HPTLC indicates that the amount of vitexin and isovitexin increased with the higher concentrations of H2O2. From this study, it can be concluded that spraying 16 and 30-mM H2O2 once a week enhances growth, mineral accumulation and stimulates bioactive compounds of the F. deltoidea plants.

Influence of Exogenous Hydrogen Peroxide on Plant Physiology, Leaf Anatomy and Rubisco Gene Expression of the Ficus deltoidea Jack var. Deltoidea

This study was carried out to investigate the regulatory effects of hydrogen peroxide (H2O2) on the growth, photosynthesis, biochemical properties, leaf anatomy and Rubisco gene expression in Ficus deltoidea var. deltoidea, a slow-growing medicinal herb. Results showed that 20-mM H2O2 treatment increased plant height, net photosynthetic rate, stomatal conductance and chlorophyll content of the plants by 10%, 20%, 127% and 57%, respectively, than a control plant. In addition, 20 mM H2O2 treatment significantly increased the carotene, total phenolic, total flavonoid and total sugar content than the control plant. The applications of H2O2 did not produce any negative effects on the leaf area, chlorophyll fluorescence, quantum yield or antioxidant activity of F. deltoidea plants. In regard to leaf anatomy, it was observed that the applications of H2O2 at 15 mM significantly improved cellular structure, leaf veins and promoted cell proliferation. Treated leaves developed a palisade layer, thickened leaf surface, the widest stomatal openings and a well-developed vascular bundle when compared to the control plant. Employing reverse transcription polymerase chain reaction (RT-PCR), the study showed that the Rubisco gene was expressed at a higher level in 15 mM H2O2 treatments than in 20 mM H2O2 treatments. The results indicate that H2O2 increased the Rubisco expression ratio up to 16-fold when compared to the untreated plants. It was conclusive that spraying 15 mM and 20 mM H2O2 twice a week enhanced growth, photosynthesis, the stomatal aperture, improved leaf anatomy and helped to regulate the expression of the Rubisco gene.

Ficus deltoidea extract down-regulates protein tyrosine phosphatase 1B expression in a rat model of type 2 diabetes mellitus: a new insight into its antidiabetic mechanism.

Ficus deltoidea var. deltoidea Jack (FD) is a well-known plant used in Malay folklore medicine to lower blood glucose in diabetic patients. For further research of the antihyperglycemic mechanisms, the protein tyrosine phosphatase 1B (PTP1B)-inhibitory effect of FD was analysed both in vitro and in vivo. To optimise a method for FD extraction, water, 50, 70, 80, 90 and 95% ethanol extracts were prepared and determined for their total phenolic and triterpene contents, and PTP1B-inhibition capacity. Among the tested extracts, 70% ethanol FD extract showed a significant PTP1B inhibition (92·0% inhibition at 200 µg/ml) and high phenolic and triterpene contents. A bioassay-guided fractionation of the 70% ethanol extract led to the isolation of a new triterpene (3β,11β-dihydroxyolean-12-en-23-oic acid; F3) along with six known compounds. In vivo, 4 weeks' administration of 70% ethanol FD extract (125, 250 and 500 mg/kg/d) to streptozotocin-nicotinamide-induced type 2 diabetic rats reversed the abnormal changes of blood glucose, insulin, total Hb, GLUT2, lipid profile, and oxidative stress in liver and pancreas. Moreover, FD reduced the mRNA expression of the key gluconeogenic enzymes (phosphoenolpyruvate carboxykinase and glucose 6-phosphatase) and restored insulin receptor and GLUT2 encoding gene (Slc2a2) expression. In addition, FD significantly down-regulated the hepatic PTP1B gene expression. These results revealed that FD could potentially improve insulin sensitivity, suppress hepatic glucose output and enhance glucose uptake in type 2 diabetes mellitus through down-regulation of PTP1B. Together, our findings give scientific evidence for the traditional use of FD as an antidiabetic agent.

Factors Affecting Cell Biomass and Flavonoid Production of Ficus deltoidea var. kunstleri in Cell Suspension Culture System

A study was conducted to establish in vitro culture conditions for maximum production of biomass and flavonoid content for Ficus deltoidea var. kunstleri, locally named as Mas Cotek, known to have a wide variety of potential beneficial attributes for human health. Size of initial inoculum, cell aggregate and initial pH value have been suggested to influent content of biomass and flavonoid for cell suspension culture in several plant species. In the present study, leaf explants were cultured by cell suspension culture procedures in MSB5 basal medium supplemented with predetermined supplements of 30 g/L sucrose, 2.75 g/L gelrite, 2 mg/L picloram and 1 mg/L kinetin with continuous agitation of 120 rpm in a standard laboratory environment. Establishment of cell suspension culture was accomplished by culturing resulting callus in different initial fresh weight of cells (0.10, 0.25, 0.50, 1.0, and 2.0 g/25 mL of media) using similar basal medium. The results showed that the highest production of biomass (0.65 g/25 mL of media) was recorded from an initial inoculum size of 2.0 g/25 mL media, whereas the highest flavonoid (3.3 mg RE/g DW) was found in 0.5 g/25 mL of media. Cell suspension fractions classified according to their sizes (500–750 µm, 250–500 µm, and <250 µm). Large cell aggregate size (500–750 µm) cultured at pH 5.75 produced the highest cell biomass (0.28 g/25 mL media) and flavonoid content (3.3 mg RE/g DW). The study had established the optimum conditions for the production of total antioxidant and flavonoid content using DPPH and FRAP assays in cell suspension culture of F. deltoidea var. kunstleri.

Blood pressure lowering effect of Ficus deltoidea var kunstleri in spontaneously hypertensive rats: possible involvement of renin-angiotensin-aldosterone system, endothelial function and anti-oxidant system.

This study investigated the effects of a standardised ethanol and water extract of Ficus deltoidea var. Kunstleri (FDK) on blood pressure, renin-angiotensin-aldosterone system (RAAS), endothelial function and antioxidant system in spontaneously hypertensive rats (SHR). Seven groups of male SHR were administered orally in volumes of 0.5mL of either FDK at doses of 500, 800, 1000 and 1300mgkg- 1, or captopril at 50mgkg- 1 or losartan at 10mgkg- 1 body weight once daily for 4weeks or 0.5mL distilled water. Body weight, systolic blood pressures (SBP) and heart rate (HR) were measured every week. 24-hour urine samples were collected at weeks 0 and 4 for electrolyte analysis. At week 4, sera from rats in the control and 1000mgkg- 1 of FDK treated groups were analyzed for electrolytes and components of RAAS, endothelial function and anti-oxidant capacity. SBP at week 4 was significantly lower in all treatment groups, including captopril and losartan, when compared to that of the controls. Compared to the controls, ACE activity and concentrations of angiotensin I, angiotensin II and aldosterone were lower whereas concentrations of angiotensinogen and angiotensin converting enzyme 2 were higher in FDK treated rats. Concentration of eNOS and total anti-oxidant capacity were higher in FDK treated rats. Urine calcium excretion was higher in FDK treated rats. In conclusion, it appears that ethanol and water extract of FDK decreases blood pressure in SHR, which might involve mechanisms that include RAAS, anti-oxidant and endothelial system.

Insulinotropic Activity of Standardized Methanolic Extracts of Ficus deltoidea from Seven Varieties.

Ficus deltoidea is a traditional medicinal plant that has been proven to show antidiabetic effects. This study focus is to assess the insulin secretion activity of Ficus deltoidea standardized methanolic extracts from seven independent varieties and mechanisms that underlie the insulin secretion action of the extracts. The cytotoxicity of Ficus deltoidea extracts was tested using viability assay. The insulin secretion assay was carried out by treating clonal BRIN BD11 cell line with standardized methanolic Ficus deltoidea extracts or glybenclamide. The clonal BRIN BD11 cell was also treated with insulin agonist and antagonist to elucidate the insulin secretion mechanism. Only the viability percentage for Ficus deltoidea var. kunstleri and intermedia was identified to be toxic at 500 and 1000 μg/ml (P<0.001). The insulin secretion for Ficus deltoidea var. deltoidea, angustifolia, and motleyana was dose-dependent; further evaluation suggested that Ficus deltoidea var. trengganuensis was involved in KATP-independent pathway. This study suggests that standardized methanolic extracts of Ficus deltoidea varieties have an insulinotropic effect on clonal BRIN BD11 cell line and can be utilized as a modern candidate of antidiabetic agents targeting the escalation for insulin secretion from pancreatic beta cells.

Discriminating Ficus deltoidea var. bornensis from Different Localities by HPTLC and FTIR Fingerprinting

A nondestructive, efficient, and accurate fingerprinting method using Fourier transform infrared spectroscopy (FTIR) has been developed and optimized for the investigation and demonstration of the variance in chemical characteristics among extracts of Ficus deltoidea Jack var. bornensis from different but closely situated origins. The capacity of attenuated total reflectance (ATR) to differentiate these samples were studied using methanol and water extracts which were preliminarily screened using HPTLC with vitexin and isovitexin being used as markers for authentication. The mobile phase used was ethyl acetate: formic acid (0.1%): methanol at ratio of 5:5:2 (v/v/v) and the profile showed that methanol extracts had higher affinity for the markers. The FTIR spectra indicated that there was no obvious difference in spectroscopic pattern for either extracts when comparing samples from different localities but the absorption intensities of some peaks were different. Multivariate statistical analyses of PCA and HCA showed that both these techniques were capable of identifying the most similar as well as most differing samples and the identification depended on the type of extract. Overall, FTIR fingerprinting has the potential to be a fast and reliable analytical methodology for the discrimination between variants of plant from closely situated locations and hence chemically similar samples.

Anti-adipogenic effects of extracts of Ficus deltoidea var. deltoidea and var. angustifolia on 3T3-L1 adipocytes

This study examined the anti-adipogenic effects of extracts of Ficus deltoidea var. deltoidia and var. angustifolia, a natural slimming aid, on 3T3-L1 adipocytes. Methanol and water extracts of leaves of the F. deltoidea varieties were analyzed to determine their total flavonoid content (TFC) and total phenolic content (TPC), respectively. The study was initiated by determining the maximum non-toxic dose (MNTD) of the methanol and water extracts for 3T3-L1 preadipocytes. Possible anti-adipogenic effects were then examined by treating 2-d post confluent 3T3-L1 preadipocytes with either methanol extract or water extract at MNTD and half MNTD (½MNTD), after which the preadipocytces were induced to form mature adipocytes. Visualisation and quantification of lipid content in mature adipocytes were carried out through oil red O staining and measurement of optical density (OD) at 520 nm, respectively. The TFCs of the methanol extracts were 1.36 and 1.97 g quercetin equivalents (QE)/100 g dry weight (DW), while the TPCs of the water extracts were 5.61 and 2.73 g gallic acid equivalents (GAE)/100 g DW for var. deltoidea and var. angustilofia, respectively. The MNTDs determined for methanol and water extracts were (300.0 ± 28.3) and (225.0 ± 21.2) µg/ml, respectively, for var. deltoidea, while much lower MNTDs [(60.0 ± 2.0) µg/ml for methanol extracts and (8.0 ± 1.0) µg/ml for water extracts] were recorded for var. angustifolia. Studies revealed that the methanol extracts of both varieties and the water extracts of var. angustifolia at either MNTD or ½MNTD significantly inhibited the maturation of preadipocytes. The inhibition of the formation of mature adipocytes indicated that leaf extracts of F. deltoidea could have potential anti-obesity effects.

Standardization and Bioassays Characterization on Malaysian herb, Ficus deltoidea Jack

Standardization of herbal materials based on their chemical and biological profile is an important prerequisite for development of herbal product. Proper identification of the source plant with related biological property is the main issue encountered in herbal standardization. Ficus deltoidea Jack (FD) is a South East Asian native plant traditionally used to treat several diseases. Pharmacological data showed that this plant exhibited good antioxidant, anti-diabetic and anti-inflammatory properties. However, there are confusion on the identification of plant material as FD occur at least in eight varieties namely Ficus deltoidea var. deltoidea, var. trengganuensis, var. kunstleri, var. montleyana, var. intermedia, var. bornensis, var. bilobata and var. angustifolia. Since reports on pharmacological and chemical studies were not at the varietal level, it is difficult to identify the best variety to be commercialized. In order to determine the most biological active variety, eight varieties of FD were collected from several localities around Malaysian Peninsular and Borneo and subjected for HPTLC and HPLC fingerprinting analysis using biologically active marker compounds, vitexin and isovitexin. HPTLC chromatograms indicated significant differences in chemical composition among varieties with var. deltoidea and var. trengganuensis possess the most chemical content. Both varieties also contain the most phenolic compounds based on Ferric-chloride assay. The concentration of marker compounds also significantly different among varieties which showed that var. trengganuensis contains the highest concentration of both markers. In vitro antioxidant and anti-diabetic data revealed the differences in biological activities which var. trengganuensis and var. motleyana showed a comparable strongest activity compared to other varieties. Finding from this study will assist the selection of the best variety to be developed as herbal product.

Antioxidant and Antihyperlipidemic Effects of the Aqueous Extract from Ficus Deltoidea var. Kunstleri Fruit and its Water Fraction IN Streptozocin-Induced Diabetic Rats

Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia; University Malaya Centre for Proteomics Research (UMCPR), Medical Biotechnology Laboratory, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; E-mail: hasni@um.edu.my