Terminalia catappa Linn. (Combretaceae)

Synonyms

Terminalia latifolia Blanco, non Swartz, Terminalia mauritiana Blanco, Terminalia moluccana Lamk., Terminalia procera Roxb.

Vernacular Names:

Malaysia: Ketapang
English: Indian Almond, Malabar Almond, Tropical Almond, Singapore almond, sea almond, wild almond, West Indian almond Barbados almond, bastard almond, Bengal almond, country almond, demarara almond, false kamani, Fijian almond, Malay almond, myrobalan, sea almond, story tree, tavola nut, tropical almond, white bombway. [1],[2],[3]

General Information

Description

Terminalia catappa is a large tree by measuring 25-40 m or 82-130 ft tall that grows in subtropical and tropical climates.  It is often associated with coastal vegetation, growing at the edges of mangrove swamps or on rocky shores.  Its main products are the nuts and timber.  It is widely planted throughout the tropics as an ornamental tree for shade, and for the edible nuts.  The nut kernel can be eaten raw.  The tree loses its leaves twice a year in most places, with the colour of the leaves turning a brilliant red to yellow before leaf shedding.

T. catappa is a large, deciduous tree with smooth grey bark and whorled branches that form a canopy and is found in tropical and subtropical regions.  The timber is moderately easy to saw and work, polishes well, has wide applications and is classed as a good constructional timber. [4] The fruit is large (2-3 inches), edible, fleshy, green (unripe) and yellow or red (when ripe).  The fruit has a husk (34.08%), a porous and fibrous pericarp (8.97%), an exocarp which is relatively thin and smooth while the hard endocarp (46.63%) enclosed an edible kernel (10.32%). [2],[4],[5]

Plant Part Used

Leaves, trunk barks, fruits. [1],[6]

Chemical Constituents

T. catappa contains hydrolyzable tannins (punicalagin as a major tannin component representing ca. 0.48% of the dry weight of the leaves, punicalin, terflavins A and B, tergallagin, tercatain, chebulagic acid, geraniin, granatin B, corilagin), flavanoids (isovitexin, vitexin, isoorientin, rutin) and triterpinoids (ursolic acid, 2α, 3β, 23-trihydroxyurs-12-en-28 oic acid and asiatic acid) but no caffeine. [1],[7],[8],[9],[10]

The bark of T. catappa yielded catappanin A, a novel complex tannin, two phenolcarboxylic acids, two phenol glucoside gallates, seven ellagic tannins, one other hydrolyzable tannin, four flavan-3-ols and two complex type tannins. [11]

The leaves contain no ascorbic acid while the contents of b-carotene, a-tocopherol and total phenols were 36.7-39.3, 0.94-1.06 and 167-198 mg/g dry weight for the green, yellow fallen and red fallen leaves, respectively. [12]

The aqueous extract from red fallen leaves contained six phenolic compounds which were identified by total ion chromatogram.  These phenols were p-hydroxybenzoic acid (15.2%), 4-hydroxyphenylpropionic acid (13.3%), m-coumaric acid (17.2%), 3,4-dihydroxybenzoic acid (17.8%), p-coumaric acid (31.4%) and and gallic acid (5.1%). [12]

The supercritical CO2 extracts of freeze-dried abscisic leaves of T. catappa contained squalene. Supercritical CO2 extraction of the freeze-dried abscisic, senescent, mature, and immature leaves and seeds of T. catappa at 40oC and 3000 psi yielded 12.29, 2.42, 1.75, 0.9, and 0% of squalene, respectively, as quantified by HPLC. [13]

The volatile components present in the supercritical CO2 (2000, 3000 or 4000 psi and 40oC) extracts of the green, yellow fallen, and red fallen leaves of T. catappa were identified using gas chromatographic retention indices, mass spectra, and authentic compounds. The aroma characteristics were analysed using gas chromatography/olfactometry.  In total, 18 compounds were identified in the leaf extracts, the three highest amounts were ethyl acetate, 6,10,14-trimethyl-2-pentadecanone and phytol.  (E,E)-2,4-Decadienal has oily odour, and, although not high in content, it was odour significant with an odour value of 18,572-31,714.  The highest odour value of 82,857-44,5715 belonged to b-ionone, the odour was described as a fruity, floral aroma. [14]

The nuts contain 22 mg of phosphorus/g dry weight, 78.14% dry weight of carbohydrates and 16.35% dry weight of crude fat. There were significant amounts of magnesium (0.4 mg/g dry weight), calcium (0.32 mg/g dry weight), iron (49.00 ug/g dry weight), zinc (0.50 ug/g dry weight), sodium (13.61 ug/g dry weight) and manganese (9.50 ug/g dry weight). The nuts also contain vitamins A (0.71 μg/g) and C (0.03 μg/g) while nitrite (1.125 μg/g) and total acidity (0.09%) were low, thus they are safe for consumption. [15]

The seed is very rich in protein (19-22%) and oil (50-52 %). [5]

Traditional Use:

The leaves, fruits and bark of T. catappa are used in dysentery, the leaves for dressing of rheumatic joints, to treat coughs and asthma. [2] The young leaves and fruits are used for headaches and colic. [16] The Sri Lankan folklore advocates the use of the juice of tender leaves of T.catappa for pains including headaches while the Sri Lankan Ayurvedic pharmacopoeia recommended different parts of T. catappa for diseases such as diarrhoea, gonorrhoea and skin ailments including scabies. [17]

The leaves are used in the treatment of leprosy and for reducing travel nausea, to get rid of intestinal parasites; treat eye problems, for wounds and to stop bleeding during teeth extraction. [2] The fruit kernel is mixed with beeswax to stop putrid exudation and bloody feces. [16] The fallen leaves are popularly used boiled or brewed in water as a drink in Taiwan, India and the Philippines to prevent hepatoma, to treat hepatitis and liver related diseases. [7],[9],[18],[19],[20]  The leaves, trunk bark and fruits were used in folk medicine for the treatment of dermatitis and for antipyretic and homeostatic purposes in India, Philippines, Malaysia, and Indonesia. [6],[18],[19],[20] The leaves are also used in the treatment of different types of cancer and to treat skin diseases. [21]

The kernel is enclosed in a fibrous shell and can be eaten raw.  It is considered to have aphrodisiac and antibacterial properties in Taiwan [15] and elsewhere where it is thought to be useful in the treatment of certain forms of sexual inadequacies such as premature ejaculation. [5]

Pre-Clinical Data

Pharmacology

Antioxidant activity

The water extract of T. catappa leaves showed antioxidant activity as evidenced its ability to inhibit Fe2+/H2O2-induced lipid peroxidation. [7]

The anti-oxidant capacities of the aqueous extracts of green, yellow fallen and red fallen leaves which were collected at three distinct periods of times: June-July, September-October and November-December were determined by 1,3-diethyl-2-thiobarbituric acid (DETBA) method, quenching of the electron paramagnetic resonance (EPR) spectrum of hydroxyl radicals reacted with the nitrone spin trap, 5,5-dimethyl pyrroline-N-oxide (DMPO), scavenging of 1,1-diphenyl-2-picrylhydryl (DPPH) radicals and superoxide anion radicals based on the reduction of nitro blue tetrazolium (NBT) and chelation of ferrous and cupric ions. [12]

The aqueous extracts from the three different leaves (1 mg/mL) showed high anti-oxidant activities in the DETBA method as they each produce 87.1-93.2% inhibition of peroxidation.  In comparison, vitamin E and butylated hydroxy anisole (BHA) at 20 mmol/L (8.6 and 3.6 mg/mL, respectively) inhibited the peroxidation of linoleic acid by 96.6% and 94.1%, respectively. The anti-oxidant activities of the three different leaves were comparable.  The anti-oxidant activities of aqueous extracts of the  three different leaves of T. catappa were comparable to those of methanolic extracts and superior to those of the essential oils (activities of 44.3, 75.3 and 80.4% for 50 mL (equivalent to 1 mg/mL) of essential oils from green, yellow fallen and red fallen leaves, respectively) and supercritical CO2 extracts (antioxidant activities of 30.7-43.9%, 63.1-76.2% and 53.7-67.7% for extracts from green, yellow fallen and red fallen leaves, respectively). The hydrolysable tannins present in T. catappa leaves were believed to be responsible for the anti-oxidant activities of the aqueous and methanolic extracts. [12]

For scavenging of hydroxyl radicals, the aqueous extract from yellow fallen leaves was more effective (75.6-81.0%) than those of aqueous extracts of green and red fallen leaves which were of similar activity (69.9-76.1% and 70.0-75.5%, respectively). The order of effectiveness for scavenging hydroxyl radicals was methanolic extracts (58.2%, 56.1% and 74.8% for 0.2 mg/mL methanolic extracts from green, yellow fallen and red fallen leaves, respectively) > aqueous extracts > essential oils (40.4-51.3% at 12.5 mL (equivalent to 0.25 mg/mL) and 69.2-82.5% at 50 mL (equivalent to 1 mg/mL).  Since scavenging of hydroxyl radicals was related to antimutagenicity, the three aqueous extracts from T. catappa leaves were postulated to possess antimutagenic properties. From the DETBA method and the hydroxyl radical scavenging method, antioxidant activities of leaves boiled for 3 mins were found to be similar to those boiled for up to 15 mins, or stirred at room temperature for 15 mins. [12]

The reducing powers of the aqueous extracts (0.5 mg/mL) from green (1.28) and yellow fallen leaves (1.15) were similar and higher than that of red fallen leaves (0.89). In comparison, the reducing powers of 0.5 mg/mL of ascorbic acid, vitamin E and BHA were 0.88, 1.15 and 1.06, respectively. At 1.0 mg/mL, the aqueous extracts from green, yellow fallen and red fallen leaves scavenged superoxide anions at 90.9, 87.5 and 72.5%, respectively, while vitamin E and BHA at 20 mmol (8.6 and 3.6 mg/mL, respectively) were without effect. The aqueous extracts of the three different leaves showed moderate scavenging of DPPH radicals.  At 50 mg/mL, scavenging abilities of the three aqueous extracts were 76.0%, 92.4% and 66.5% for green, yellow fallen and red fallen leaves, respectively, while those of 0.1 mg/mL vitamin E and BHA were 95.2% and 95.5%, respectively.  Better scavenging of DPPH radicals were shown by methanolic extracts than the aqueous extracts.  Aqueous extracts from green leaves were better than red fallen and yellow fallen leaves at scavenging superoxide anion radicals and DPPH radicals. The aqueous extracts (5 mg/mL) from green, yellow fallen and red fallen leaves showed excellent chelation of ferrous ions at 99.2%, 99.0% and 84.0%, respectively.  The same concentration of aqueous extracts for green, yellow fallen and red fallen leaves showed good chelation of cupric ions at 28.8%, 25.8% and 18.0%, respectively.  In comparison, EDTA (1 mg/ml) chelated both ferrous and cupric ions at 99.5%.  The green leaves contain high amounts of tannins which contribute to the astringent taste thus making the green leaves unpalatable. People prefer to use fallen leaves of T. catappa as senescence of the leaves decrease their content of tannins. [12]

The green, yellow fallen and red fallen leaves of T. catappa were extracted using dichloromethane, ethyl acetate, methanol and n-pentane, and the antioxidant activities of these various solvent extracts determined via measurement of the reducing power, scavenging of DPPH and hydroxyl radicals, and chelating effect on ferrous ions. For detection of hydroxyl radicals, DMPO was used as the radical trap and the resultant DMPO-OH adducts was detected with an EPR spectrometer.  The reference anti-oxidants were a-tocopherol, ascorbic acid and butylated hydroxyanisole.  The yield of oleoresins with the four different solvents were consistently in the order of yellow fallen (6.34-10.50%)>red fallen (5.12-9.98%)>green leaf extracts (2.36-6.08%).  The higher yields were obtained with ethyl acetate or methanol extraction for the three different leaves.  The anti-oxidant activities by the DETBA method for all three leaves were higher with the oleoresins extracted with the more polar solvents being in the order of methanol>ethyl acetate> dichloromethane>pentane extracts.  At concentrations below 0.1–0.5 mg/mL, the antioxidant activity of each of the extracts increased with the concentration but at concentrations which were higher than this range, the antioxidant activity was decreased with an increase in concentration.  These results indicate that at higher concentrations, the solvent extracts showed a prooxidant effect.  The methanol was the most effective solvent for the extraction of antioxidant components from the three different leaves as their methanolic extracts showed the highest antioxidant activities. [9]

The three methanolic extracts showed excellent reducing powers at concentrations as low as 0.5 mg/mL but at 0.1 mg/mL, the three methanolic extracts showed reducing powers in the range of 0.48-0.66, which were comparable to that of vitamin E (0.45), though much less than those of BHA and vitamin C (0.98 and 1.04, respectively).  The three methanolic extracts showed excellent scavenging of DPPH radicals at concentrations as low as 0.1 mg/mL in the range of 92.5-95.7% which was comparable to those of vitamins C and E and BHA (95.2-96.7%).  The methanolic extracts from green, yellow fallen and red fallen leaves (0.2 mg/mL), scavenged hydroxyl radicals by 58.2, 56.1 and 74.8%, respectively.  In comparison, vitamin E (20 mM or 8.6 mg/mL) showed a scavenging ability of 43.8%.  Since the ability to quench hydroxyl radicals was directly related to antimutagenic potential, it was anticipated that the three methanolic extracts of T. catappa leaves may possess antimutagenic properties.  The methanolic extract from green leaves showed better chelation of ferrous ions than those from yellow fallen and red fallen leaves.  At measure 30 mg/mL concentrations, methanolic extracts from green, yellow fallen and red fallen leaves chelated ferrous ions by 77.3, 48.6 and 48.3%, respectively.  In contrast, EDTA (1 mg/mL) chelated ferrous ions by 99.5%. [9]

The supercritical CO2 extracts of freeze-dried abscisic leaves of T. catappa exhibited potent anti-oxidative (in an iron/ascorbate system with linoleic acid, and in a pork fat storage system for inhibition of conjugated diene hydroperoxide formation) and DPPH scavenging activities which increased with leaf maturity. The seed extract showed potent inhibition of conjugated diene hydroperoxide formation but had little DPPH scavenging activity. [13]

The punicalagin and punicalin were the most abundant tannin components of T. catappa. They showed the strongest anti-oxidative effects of the group of tannins of T. catappa.  The antioxidant activity was evaluated by determination of their ability to prevent lipid peroxidation, inhibit superoxide formation and by their free radical scavenging activity. [22]

 

 

The mitomycin C-induced micronuclei in Chinese hamster ovary K1 (CHO-K1) cells was significantly suppressed when the cells were simultaneously treated or pretreated with the aqueous extract of T. catappa leaves (75 & 150 µg/mL). This inhibition was not seen when CHO-Kl cells were treated with 3OO µg/mL of T. catappa extract.  The survival of CHO-Kl cells were unaffected by treatment with 75 and 150 µg/mL of T. catappa extract.  At 3OO µg/mL, treatment with T. catappa for 1 or 6 hours reduced the survival fraction of CHO-Kl cells to 90 and 60% of the control value, respectively.  The administration of T. catappa extracts (4.8 & 24 mg/animal/day by gastric intubation) for 8 days to male mice resulted in a reduction in mitomycin C-induced micronuclei in peripheral blood.  The anticlastogenic effects of T. catappa may be due to its anti=oxidant potential as T. catappa dose-dependently inhibited lipid peroxidation in vitro with an IC50 value of 7.4 µg/mL.  In human mononuclear leukocytes, T. catappa extract reduced 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced production of hydrogen peroxide.  This effect was attibuted to T. catappa’s ability to inhibit TPA induction of hydrogen peroxide formation and/or to scavenging of oxygen free radicals in human leucocytes. The T. catappa extract used here contained punicalin (0.15%) and punicalagin (0.027%) by HPLC analysis. [8]

The supercritical CO2 (2000, 3000 or 4000 psi and 40oC) extract of the green, yellow fallen, and red fallen leaves of T. catappa were tested for antioxidant activity by the DETBA method.  The higher inhibition of peroxidation was elicited by extracts from yellow fallen and red fallen leaves than from green leaves regardless of the pressures employed.  The supercritical CO2 extraction at 2000 psi and 40oC produced extracts with better antioxidant activity while lower inhibition of peroxidation was seen when the extracts prepared by extraction at 4000 psi and 40oC.  However, in comparison to inhibition of peroxidation by a-tocopherol (96% inhibition at 20 mM or 8.6 mg/mL), the anti-oxidant activities of these extracts were low. [14]

 

 

Antidiabetic activity

The aqueous and cold extracts of the fresh tender leaves of T. catappa showed antidiabetic effects in alloxan-induced diabetic rats. The aqueous extract was obtained by extraction of powdered leaves with distilled water in a Soxhlet apparatus for 18 hours while the cold extract was prepared by extraction with distilled water for 7 days with daily stirring for 2 hours with a mechanical stirrer using chloroform as the preservative.  The wistar albino rats (150-200 g) were made diabetic by a single intraperitoneal injection of alloxan monohydrate (150 mg/kg) and treatment with T. catappa extracts at doses which correspond to 1/5th their lethal doses was initiated 48 hours later. The glibenclamide was used as the control antidiabetic agent. Three weeks of daily treatment with the aqueous extract (43 g/kg per day p.o.) or with the cold extract (46 mg/kg per day p.o.) elicited a dose-dependent drop in blood sugar levels by 25-62% with the maximum effect attained after 15 days and remaining at the same level in the third week.  The extracts also reversed alloxan-induced deceases in the body weights of rats beginning from the 7th day of treatment. Both extracts reversed the elevations in the levels of serum cholesterol, serum triglycerides, serum low density lipoprotein (LDL), serum creatinine, serum urea and serum alkaline phosphatase that were seen in alloxan-induced diabetic rats. The decrease in serum high density lipoprotein (HDL) levels that was induced by alloxan was reversed by both extracts. Histologically, both extracts produced regeneration of the β-cells of the pancreas which were previously necrosed by alloxan.  The regeneration was comparable to that produced by glibenclamide.  The antidiabetic effect of T. catappa was attributed to its β-carotene content. [1]

The antidiabetic effects of the petroleum ether, methanol, and aqueous extracts of the fresh, unriped, green fruits of T. catappa which were collected in October, were determined in alloxan diabetic rats. The diabetes was induced in rats by a single intraperitoneal injection of alloxan monohydrate (150 mg/kg).  Forty-eight hours later, the animals were treated with the plant extracts and blood samples withdrawn weekly for blood glucose estimation for the duration of the 3-week study.  The extracts elicited significant antidiabetic effects at doses which were 1/5th of their lethal doses, i.e. 68, 40 and 42 mg/kg per day p.o. for the petroleum ether, methanol and aqueous extracts, respectively.  T. catappa extracts cause a dose-dependent fall in blood sugar levels by 25–62% with the maximum effect seen after 15 days of treatment.  The methanol and aqueous extracts of T. catappa elicited significant anti-hyperglycemic activity in rats without significantly affecting their body weights.  These extracts also improved the lipid profile of the diabetic rats. The methanolic and aqueous extracts produced regeneration of the pancreas of diabetic animals which were previously necrosed by alloxan, comparable to the effects of the standard antidiabetic drug, glibenclamide.  This effect was postulated to be due to b-carotene, which is present in the fruit. [23]

Anticancer activity 

The ability of the water extract of T. catappa to prevent metastasis was investigated in vitro, using A549 cell line, a highly metastatic human lung cancer cells, and in vivo, using Lewis lung carcinoma (LLC)-bearing mice, an established animal model for metastasis. The water extract (0-100 µg/mL) did not affect the viability of A549 cells although it was cytotoxic to LLC cells in a concentration-dependent manner (IC50 of 14.5 µg/mL).  The invasion and motility of A549 cells was significantly reduced by the water extract (50-100 µg/mL) in a concentration-dependent manner.  After 24 hours, the extract (100 ug/mL) left only 24.8% and 28.8% of cells remaining, for cell invasion and motility, respectively.  A similar inhibitory effect was seen in LLC cells treated with the extract for 24 hours.  The levels of the specific endogenous inhibitors of proteolytic enzymes, TIMP-2 and PAI-1, were gradually decreased by the water extract (10-100 µg/mL) in both A549 and LLC cancer cells.  Although the relationship between proteolytic enzymes (i.e. MMP-2, MMP-9, and µ-PA activities) and their endogenous inhibitors (i.e. TIMP-2 and PAI-1) on anticancer metastasis has not been confirmed with certainty, several studies have shown that anti-lung cancer metastasis could be due to lowered levels of proteolytic enzymes with the levels of their endogenous inhibitors remaining unchanged or decreased.  In vivo, the water extracts decreased lung metastases of LLC-bearing C57BL/6 mice by 68% compared to controls.  After 30 days of treatment with the water extract, there was a 2.6-fold reduction in small solid tumors in tumour-bearing mice as compared to controls.  At this time, the tumor weight was reduced by 2.3-fold and there was no apparent signs of toxicity as indicated by body weight monitoring (Chu et al, 2007).   These results indicate that the water extract of T. catappa is a potentially important agent for the prevention of lung cancer metastasis. [24]

Both the water extract of T. catappa leaves (25-00 µg/mL) and its major tannin component, punicalagin (1.0 µg/mL) significantly protected CHO-K1 cells against bleomycin-induced hgprt gene mutation frequency when the cells were pretreated with the extract or with punicalagin for 24 hours prior to exposure to bleomycin (2.25 mU/mL) for another 24 hours. The punicalagin was not as effective as the extract since the most effective anti-mutagenic doses of the extract, 50 and 75 µg/mL, contained 0.24 and 0.36 µg/mL of punicalagin, respectively.  Similarly, CHO-K1 cells were also protected against bleomycin-induced DNA-strand breaks, measured by the comet assay, when the cells were pretreated with the extract (75 and 100 µg/mL, equivalent to 0.36 and 0.48 µg/mL of punicalagin, respectively) or with punicalagin (0.05-1.0 µg/mL) for 24 hours before exposure to bleomycin (15 mU/mL) for 2 hours. These concentrations of either the extract or punicalagin were non toxic to CHO-K1 cells as cellular viability were not affected by maximum concentrations of 100 µg/mL of the extract or 1.0 µg/mL of punicalagin for 24 hours.  The stong anti-genotoxic effects of the extract and punicalagin were attributed to their ability to ameliorate bleomycin-induced reactive oxygen species formation which was responsible for bleomycin’s DNA-damaging effect.  Both the extract and punicalagin suppressed the intracellular formation of superoxides and hydrogen peroxides by bleomycin, probably through direct scavenging of superoxide anions and H2O2. [6]

To study the chemopreventive effects of T. catappa water extract and punicalagin, H-ras-transformed NIH3T3 cells were used as these cells are known to express typical phenotypes of malignant transformation, viz, alterations in cell morphology, increased growth rate, reduced serum dependence, loss of density-dependent growth inhibition, acquisition of anchorage-independent growth potentials and the ability to form tumors in experimental animals. T. catappa water extract suppressed the growth of H-ras-transformed NIH3T3 cells in a concentration-dependent manner.  Cellular growth was completely suppressed by 100 µg/mL of the water extract although in non-transformed NIH3T3 cells, this concentration only produced 30% cell death (Chen & Li, 2006).  The punicalagin also inhibited the growth of H-ras-transformed NIH3T3 cells in a concentration-dependent manner.  Almost complete inhibition was achieved at 50 µg/mL although in non-transformed NIH3T3 cells, the growth rate at this concentration was at 70%.  The IC50 values for the water extract and punicalagin on H-ras-transformed NIH3T3 cells were 45 and 17 µg/mL, respectively.  Both compounds did not increase the number of dead cells.  This indicate that they work through inhibition of cell proliferation via a concentration-dependent induction of G1 arrest of H-ras-transformed NIH3T3 cells as both elevated the population of G1 cells.  Subtoxic concentrations of the water extract (5 & 10 µg/mL) and punicalagin (5 µg/mL) suppressed anchorage-independent cell growth, a malignant phenotype of H-ras-transformed NIH3T3 cells. [7]

Higher concentrations of the water extract (100 µg/mL) or punicalagin  (25 µg/mL) completely inhibited anchorage-independent cell growth.  Neither Ras protein level nor Ras activity was affected by punicalagin  (5-50 µg/mL).  The elevated superoxide levels that was seen in H-ras-transformed NIH3T3 cells was lowered by punicalagin (5-50 µg/mL) without any effects being produced on nontransformed NIH3T3 cells. The punicalagin (25 & 50 µg/mL) inhibited phosphorylated JNK-1 without affecting JNK-1 protein contents, which could be attributed to its suppression of Ras-induced superoxide generation.  JNK is essential for Ras-induced cellular transformation.  The reduction of JNK-1 activity in H-ras-transformed NIH3T3 cells may contribute to the inhibition of cell growth and anchorage-independent growth by punicalagin.  Similarly, phosphorylated p38 was suppressed by punicalagin (5-50 µg/mL) while p38 protein contents were not affected. There was no effect of punicalagin on phosphorylated ERK1/2.  In contrast, punicalagin did not affect JNK-1 and p38 phosphorylation in non-transformed NIH3T3 cells. [7]

The hot water extract of T. catappa showed potent short-term chemopreventive action on biomarkers of colon carcinogenesis. Colon cancer was induced in 6 weeks old male F344 rats by weekly subcutaneous (s.c.) injections of azoxymethane (20 mg/kg body weight) for 2 weeks.  The rats were fed a diet containing 0.02 and 0.1% T. catappa for 5 weeks beginning a week before the first injection of azoxymethane.  T. catappa (0.02 & 0.1%) significantly reduced the number of aberrant crypt foci (ACF)/colon/rat and b-catenin accumulated crypts/cm2/rat when compared to the control.  ACF is well accepted as visible preneoplastic lesions that develop in the colonic mucosa of rats treated with azoxymethane, thus it is a useful biomarker for colon carcinogenesis. T. catappa also significantly reduced cell proliferation activity of colonic mucosal epithelium as the proliferating cell nuclear antigen index was lower than that of the control. The protection afforded by T. catappa against colon carcinogenesis waspostulated to be related to its antioxidant activity. [18]

The supercritical CO2 extract of T. catappa leaves was not mutagenic at a dose of 0.5 mg/plate in the Ames test nor was it toxic. The decreases in the temperature (60, 50, and 40oC) and pressure (4000, 3000, and 2000 psi) used for extraction increased the antimutagenic activity of supercritical CO2 extracts with the most potent antimutagenic effects observed in extracts obtained at 40oC and 2000 psi.  The extract (0.5 mg of extract/plate), with S-9, inhibited approximately 80% of the mutagenicity of benzo[a]pyrene while the mutagenicity of N-methyl-N'-nitroguanidine (without S-9) was inhibited by 46%.  The supercritical CO2 extract (0-500 µg/mL) elicited dose-dependent growth inhibition of both human hepatoma (Huh 7) and normal liver (Chang liver) cells but cytotoxicity of the extracts to the hepatoma cells was greater than to normal liver cells. [25]

Antiviral activity

The punicalin and punicalagin inhibited HIV replication in infected H9 lymphocytes with little cytotoxicity, and inhibited purified HIV reverse transcriptase with ID50 values of 8 and 5 µM, respectively. [8] The chebulagic acid and punicalin blocked the binding of recombinant HIVcoat protein gp120 (rgp120) to its normal cellular receptor, CD4 (From Liu et al, 1996).  The fruit of T. catappa contains ellagic-acid which has anti-HIV activity. [23]

Antifungal and antimicrobial activity

The methylene chloride and methanol extracts of T. catappa showed antifungal activity against Pythium ultimum, Rhizoctonia solani, Sclerotium rolfsii, Aspergillus fumigatus and Phytophthora parasitica. The methanol extract showed marked antifungal activity against Pythium ultimum and Phytophthora parasitica. [21]

The antibacterial effects of the petroleum ether, chloroform and methanol extracts of the dried roots of T. catappa were determined by the cup plate agar diffusion method.  The petroleum ether extract was devoid of antimicrobial activity.  Both the chloroform and methanol extracts showed good antimicrobial activity against Gram positive and Gram negative microorganisms. Compared to the other extracts, the chloroform extract showed prominent antimicrobial activity against S. aureus (MIC of 0.4 mg/mL) and E. coli.  The methanol extract exhibited potent activity against E. coli (MIC of 0.065 mg/mL). [26]

Anti-inflammatory activity

The ethanolic extract of T. catappa leaves showed anti-inflammatory activity in acute and chronic mouse models of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema. The activity was attributed to ursolic and 2a,3b,23-trihydroxyurs-12-en-28-oic acids which were isolated from the chloroform fraction.  In the acute model, the chloroform fraction (1 mg/ear) exhibited significant anti-inflammatory activity which was comparable to that of indomethacin (0.3 mg/ear).  The ED50 for anti-inflammatory activity of the chloroform fraction was 0.86 mg/ear.  The ethyl acetate fraction showed mild antiedema effect, while the crude ethanol extract showed little or no anti-inflammatory effect.  Both ursolic acid and 2a,3b,23-trihydroxyurs-12-en-28-oic acid produced over 50% reductions in edema in the acute model.  In the chronic ear edema model, the crude ethanolic leaf extract (1 mg/ear) and the chloroform fraction reduced ear edema by 32.0 and 66.0%, respectively, while ursolic and 2a,3b,23-trihydroxyurs-12-en-28-oic acids (each at a dose of 0.3 mg/ear) reduced ear edema by 96.9 and 94.8%, respectively.  The chloroform fraction and the two isolated compounds produced a concomittant decrease in neutrophil infiltration as detected by reductions in myeloperoxidase activities by 59.0%, 92.6% and 90.9%, respectively.  In contrast, the crude ethanolic leaf extract reduced myeloperoxidase activity by 17.6%. [20]

Analgesic activity

The analgesic, antihyperalgesic and anti-inflammatory activities of the extracted juice from the tender leaves of T. catappa were investigated in rats. The extracted juice was produced by maceration of the tender leaves in a mortar (40% v/w; 2.5 g of leaves produced 1mL extract).  The extract was orally administered to male rats in doses of 5, 10, or 15 mL/kg while female rats at different stages of the estrous cycle were given a 10 mL/kg dose.  The analgesic potential was determined at 1, 3 and 5 hour.  Other groups of rats were given the extract (10 mL/kg) and were subjected to carrageenan-induced paw edema, inflammatory and formalin-induced pain tests.  In the hot plate test, the higher doses (10 and 15 mL/kg) increased the reaction time and changed the percent maximum possible effect at 3 hours post treatment.  IC50 values for the extract and for indomethacin (the reference analgesic agent) in the hot plate test were 7.8 mL/kg and 3.37 mg/kg, respectively.  No analgesic activity was shown by all doses of the extract in the tail flick test.  These results suggest that antinociception by the extract was mediated supraspinally as the hot plate test largely measures supraspinally organised responses while tail flick test predominantly measures spinal reflexes.  The extract (10 mL/kg) also produced significant analgesia in female rats which was not affected by the estrous cycle.  Antinociception by the extract was not antagonised by naloxone nor by metochlopramide.  The onset of the analgesic action was slow (3 hours) and of short duration (reversible by 5 hours).  Thus for use as an analgesic, it is only recommended for mild to moderate pain.  The extract did not induce sedation.  Neither anti-inflammatory nor antihyperalgesic activities were exhibited by the extract in the carrageenan-induced paw edema model.  Pain in the early phase of the formalin pain test was significantly reduced by the extract but no reduction was seen in the late phase. [17]

Hepatoprotective/antioxidant activity

The water extract of T. catappa markedly scavenged DPPH and superoxide anion (O2- ) radicals with IC50 values of 0.85 µg/mL and 0.2 µg/mL, respectively. Chebulagic acid and corilagin present in both the water extract and in the 50% ethanol extract showed strong radical scavenging activity.  The IC50 values for scavenging of O2- were 0.84 µM and 0.32 µM for chebulagic acid and corilagin, respectively.  For scavenging of peroxyl radicals, the IC50 values were 14.5 µM and 16.4 µM for chebulagic acid and corilagin, respectively.  Reactive oxygen species (ROS) generation from PMA-stimulated leukocytes was inhibited more by corilagin (IC50 of 69 µM) than by chebulagic acid (IC50 of 154 µM).  Both compounds markedly inhibited enzymatic and non-enzymatic lipid peroxidation with IC50 values of 1.6-4.5 mM) (Kinoshita et al, 2007). An animal model of fulminant hepatic failure with predominant hepatocyte apoptosis with caspase-dependency was produced by treatment of rats with D-Galactosamine (D-GalN)/lipopolysaccharide(LPS) (600 mg/kg GalN, s.c. and 0.5 mg/kg LPS i.p.).  Administration T. catapa extract (1 mg/kg, i.p.) at 1 and 15 hours before treatment of rats with GalN/LPS resulted in significant falls in the serum levels of liver marker enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and serum glutathione S-transferase (GST) activities which were elevated by GalN/LPS. The extract alone did not alter serum AST and ALT levels of control rats.  Pretreatment of GalN/LPS-treated rats with the extract also resulted in reversals in GalN/LPS-induced increase in lipid peroxide level in the liver homogenate, GalN/LPS-induced increase in nitric oxide (NO) levels in the liver and serum, and less loss of cytosolic GST.  Pretreatment with the extract also abolished the increase in caspase 3 activity and DNA fragmentation that were observed in the livers of GalN/LPS-treated rats.  Free radical formation, specifically hydroxyl and singlet oxygen was seen in GalN/LPS-treated rat liver, these were abolished by pretreatment with the extract.  Corilagin produced similar effects as the water extract in GalN/LPS-treated rats. Administration of corilagin (1 mg/kg, i.p.) at 1 and 15 hours before GalN/LPS-treatment in rats also led to reversals in GalN/LPS-induced increases in serum ALT, AST, GST levels. Corilagin also reversed GalN/LPS-induced increases in liver lipid peroxide levels, chromosomal DNA fragmentation and GalN/LPS-induced increase in caspase 3 activity. [10]

The extract of T. catappa leaves protected mice against D-GalN-induced liver injury.  D-GalN can induce acute hepatitis in mice which closely resembles human viral hepatitis both in the morphological and functional changes.  In this study, mice were pretreated with T. catappa leaf extract (20, 50 and 100 mg/kg/d, p.o.) for 7 days prior to treatment with D-GalN (800 mg/kg, i.p.).  The extract was able to completely block the increase in serum ALT activity caused by D-GalN, denoting hepatoprotection as the ALT enzyme is an index for cell membrane damage.  In vitro, T. catappa leaf extract (0.1, 0.5 and 1.0 mg/mL) protected against D-GalN-induced cytotoxicity in primary cultured hepatocytes from fetal mice in a dose-dependent manner.  The extract at 1.0 mg/mL concentration almost totally block D-GalN-induced cytotoxicity.  T. catappa extract also protected against D-GalN-induced increase in AST level and D-GalN-induced decrease in superoxide dismutase (SOD) activity.  AST is an index of mitochondrial damage as mitochondria contains 80% of the enzyme.  The effects of 2a, 3b, 23-trihydroxyursane-12-en-28-oic acid, a compound isolated from T. catappa leaf, were studied to determine the mechanism for the hepatoprotection by the extract.  2a, 3b, 23-Trihydroxyursane-12-en-28-oic acid (50, 150 and 500 µmol/L) showed dose-dependent inhibition of Ca2+-induced mitochondrial swelling and dose-dependent scavenging of superoxide radicals.  Thus, hepatoprotection against D-GalN-induced hepatotoxicity may be attributed to direct mitochondrial protection and to scavenging of reactive oxygen species by the extract’s components. [27]

The protective effects of chloroform extracts of T. catappa leaves on carbon tetrachloride (CCl4)-induced liver damage were investigated in mice. The hepatic damage was induced with 0.15% CCl4 in olive oil (10 mL/kg, i.p.).  Oral pretreatment of the mice for 5 days with 20, 50 or 100 mg/kg of the chloroform leaf extract significantly inhibited CCl4-induced increases in the activities of serum AST and ALT and the CCl4-induced increase in liver lipid peroxidation in a dose-dependent manner.  The highest dose of the extract (100 mg/kg) almost completely blocked the changes elicited by CCl4.  The hepatoprotective effects of T. catappa chloroform leaf extract were confirmed morphologically.  CCl4 caused massive fatty change, gross necrosis, broad infiltration of lymphocytes and Kupffer cells around the central vein and loss of cellular boundary in the liver.  Pretreatment with the extract prevented against CCl4-induced changes as the livers of pretreated mice showed mild degrees of fatty change, necrosis and lymphocyte infiltration.  T. catappa chloroform leaf extract showed protective activities against liver mitochondrial damage as pretreatment with the extract effectively prevented CCl4-induced disruption of liver mitochondrial membrane potential, intramitochondrial Ca2+ overload and CCl4-induced suppression of mitochondrial Ca2+-ATPase activity.  Dimethyl diphenyl bicarboxylate (200 mg/kg daily for 5 days, p.o.) was the positive reference agent. [19]

The pretreatment of mice with the chloroform fraction of the ethanol extract of T. catappa leaves (10 or 30 mg/kg, intragastric administration) protected mice from CCl4 toxicity. CCl4-induced elevations in the activities of serum ALT and serum AST were reversed by T. catappa extract. CCl4-induced over-transcription of IL-6 gene was markedly suppressed by T. catappa which also blocked the expression of IL-6 protein especially in the area surrounding the terminal hepatic vein. [28]

In a similar study, the hepatoprotective effects of T. catappa chloroform extract against CCl4-induced liver injury in mice and its effects on IL-6 gene overexpression were determined.  Mice were pretreated with the extract (20, 50, 100 mg/kg/d) for 5 days followed by an intraperitoneal injection of CCl4.  The extract dose-dependently reversed the increase in serum AST caused by CCl4 and decreased CCl4-induced increase in IL-6 mRNA level in the liver.  At a high dose (100 mg/kg), the extract was able to lower the IL-6 mRNA level to control values.  Histological changes due to CCl4 treatment such as the infiltration of inflammatory cells and hepatocyte swelling, were significantly reduced by pretreatment of mice with the extract. [29]

The crude aqueous extract of T. catappa protected rat liver against against CCl4-induced hepatotoxicity.  The extract also showed antioxidant activity in FeCl2-ascorbic acid induced lipid peroxidation in rat liver homogenate.  Superoxide anion radical scavenging by T. catappa was shown using electron spin resonance and spintrapping techniques. [30]

Aphrodisiac activity

A 4.3 kDa bioactive peptide from Eurycoma longifolia, a plant with reputed aphrodisiac activity, was found to increase testosterone levels in rat Leydig cells.  However, the 4.3 kDa bioactive peptide peak was not detected in T. catappa even though it reportedly prolonged the ejaculatory latency, thus suggesting a dissimilar action of aphrodisiac activity from E. longifolia. [16]

The kernel of T. catappa seeds exhibited aphrodisiac activity in rats. The seeds were obtained from ripe and partially dried fruits that have gray coloured pericarp.  A suspension of the seed kernel (1500 mg/kg or 3000 mg/kg) in 1% methyl cellulose was orally administered to male rats and their sexual behaviour with a receptive female was monitored 3 hours later. Another group of rats was administered 3000 mg/kg of the seed suspension daily for 7 days.  Effects on sexual behaviour and fertility were evaluated by overnight pairing with a pro-oestrous female.  The assessment was carried out on days 1, 4 and 7 of treatment and day 7 post-treatment.  The lower dose (1500 mg/kg) impaired sexual arousability, as there was prolongation of the time taken to mount, intromit or ejaculate without any effect on libido, sexual vigour or sexual performance.  The prolongation of ejaculatory latency suggests an aphrodisiac action.  Libido, sexual vigour and sexual performance were unimpaired during the aphrodisiac action as all treated rats mounted and intromitted without inhibition of mount-and-intromission frequencies or copulatory efficiency or intercopulatory interval.  The higher dose (3000 mg/kg) reversibly inhibited all parameters of sexual behaviour except the frequencies of mounting and intromission, and copulatory efficiency.  The inhibitory effect on sexual behaviour had a rapid onset and recovery.  The inhibition that was seen with the higher dose of T. catappa seed suspension was due to sedation.  Both doses of T. catappa seed did not reduce blood testosterone levels. [31]

Anti- sickling of human ‘SS’ erythrocytes activity

T. catappa leaves showed potential for the management of sickle cell disease ((HbSS). The ethanol extract of the leaves dose-dependently inhibited osmotically-induced hemolysis of human erythrocytes and prolonged the clotting time of uncoagulated blood. [32]  The extract (1.0 mg/mL) prevented and reversed metabisulphite-induced sickling of human ‘SS’ erythrocytes.

Other uses

The fruit shell of T. catappa proved to be a useful precursor material for the preparation of a carbon adsorbent used in the removal of Hg(II) from aqueous solutions. The adsorption capacity of the carbon was 94.43 mg/g at pH 5.0 for particle sizes of 180-210 µm. [4]

The acidified methanol extract of the outer covering of T. catappa fruits is a useful, economical, simple and accurate indicator for acid-base titrations. [2]

Toxicities

The oral LD50 doses of the aqueous and cold extracts of T. catappa leaves in mice were 215 and 230 mg/kg, respectively. [1] The oral LD50 doses of the petroleum ether, methanol, and aqueous extracts of T. catappa fruits in mice were 343, 195, and 210 mg/kg, respectively. [23]

T. catappa seed suspension (1500 mg/kg) did not produce liver toxicity in male rats but a higher dose (3000 mg/kg) caused moderate elevations in serum AST and ALT activities indicating liver toxicity. [31] Both doses of T. catappa seeds did not elicit general toxicity nor haemotoxicity, stress, muscle deficiency, muscle incoordination, analgesia or hypoglycaemia.  T. catappa tender leaf extract (5, 10, or 15 mL/kg) was well tolerated in male and female rats and produced no overt signs of toxicity, stress or behavioural abnormalities. [17]

T. catappa (0.02 & 0.1% in diet for 5 weeks) produced no significant effects on the body, liver and relative liver weights of male rats given. [18]

Adverse effects of defatted T. catappa seed meal

 

The effects of defatted T. catappa seed meal on the growth performance and carcass of 3-weeks old weaning albino rats (average weight of 24.0±3.4 g) fed on this diet for six weeks were determined in comparison to rats fed a soybean meal (control).  Defatted T. catappa seed meal caused depression in the growth rate, enlargement of rat tissues with adverse effects on carcasses of the rats.  Defatted T. catappa seed meal significantly reduced the rats’ body weights to one-third of those of controls while causing a significant increase in the organ (brain, liver, kidney, stomach, small intestine) to body weight ratio in comparison to controls.  The carcasses of rats fed defatted T. catappa seed meal had significantly reduced ether extract, crude protein and ash contents but significantly higher crude fibre and the nitrogen free extract when compared with controls.  Two of the 6 rats fed the defatted T. catappa seed meal died before the end of the experiment indicating possible toxicity of defatted T. catappa seed meal on the internal organs of the rats which were significantly heavier than those of controls as indicated by the higher organ to body weight ratio in the former group.  The seeds of T.catappa contain inherently high protein quality, but this was not utilised to form body muscle possibly as a result of chelation by antinutrients present in the seed especially phytates, insoluble oxalates and condensed tannins.  Ether extract of the carcass of rats fed the T.catappa seed meal was significantly reduced indicating that lipid metabolism in these animals was adversely affected. [5]

Clinical Data

Clinical Trials

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Adverse Effects in Human:

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Use in Certain Conditions

Pregnancy / Breastfeeding

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Age Limitations

Neonates / Adolescents

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Geriatrics

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Chronic Disease Conditions

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Interactions

Interactions with drugs

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Interactions with Other Herbs / Herbal Constituents

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Contraindications

Contraindications

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  1) Botanical Info

References

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