Sonneratia apetala (Buch.-Ham.) Fruit Extracts Ameliorate Iron Overload and Iron-Induced Oxidative Stress in Mice.

Iron overload results in oxidative damage to various biomolecules including DNA, proteins and lipids which ultimately leads to cell death. The Sonneratia apetala fruit contains a high content of antioxidants and displays several bioactive properties. Therefore, the powder of the S. apetala fruit was successively fractionated into n-hexane (Hex), chloroform (Chl), and methanol (Met) fractions to evaluate their efficiency in ameliorating iron overload. In vitro, a colorimetric method was used to assess the Fe-chelating activity of the fractions using ferrozine. The fractions were also used in vivo to examine their efficacy in ameliorating iron overload and iron-induced oxidative stress in mice induced by intraperitoneal injection of ferric carboxymaltose at 100 mg/kg body weight (bw). Among the fractions, Met showed the highest Fe-chelation ability with an inhibitory concentration 50 of 165 µg/mL followed by Hex (270 µg/mL), and Chl (418 µg/mL). In vivo, the results showed a significantly (P<0.05) lower iron profile (iron and ferritin concentrations in serum and liver tissue and total iron-binding capacity of serum) in the Met and the Hex treated mice groups than in the iron-overloaded group. Met at 1,000 µg/kg bw completely ameliorated iron overload in the blood and the liver tissue of mice. At this concentration, Met also prevented iron-induced oxidative stress in the liver tissue of iron-overloaded mice by restoring reducing power, total antioxidant capacity, and total protein. Thus, the S. apetala fruit, especially its Met fraction can be used in treating iron overload and associated toxicity.

Anti-Bacterial, Anti-Diarrheal, and Cytotoxic Activities of Edible Fruits in the Sundarbans Mangrove Forest of Bangladesh.

This study evaluated anti-bacterial, anti-diarrheal, and cytotoxic activities of ten edible fruits (Aegiceras corniculatum, Avicennia officinalis, Bruguiera gymnorrhiza, Ceriops decandra, Heritiera fomes, Nypa fruticans, Phoenix paludosa, Sarcolobus globosus, Sonneratia caseolaris, and Xylocarpus mekongensis) in the Sundarbans mangrove forest of Bangladesh. Highest antibacterial activity was shown for ethanol:methanol (1:1) extracts of S. caseolaris (2 mg/disc), which demonstrated inhibition zones of 16.7, 17.0, 14.7, 15.7, and 15.7 mm against Escherichia coli, Klebsiella sp., Shigella boydii, Shigella sonnei, and Staphylococcus aureus, respectively. In mice with castor oil-induced diarrhea, S. caseolaris extract (250 mg/kg body weight) showed the highest inhibition (87.7%) and delayed the onset time (273 min) of diarrheal episodes, followed by A. corniculatum extract (inhibition, 83.6%; delayed onset time, 187.4 min). S. caseolaris and A. corniculatum fruit powders were successively fractionated into n-hexane, diethyl ether, chloroform, ethanol, and methanol. Antidiarrheal activity significantly increased with increasing polarity of these fractions. In brine shrimp lethality assay, S. globosus extract demonstrated the highest cytotoxicity (59.2 µg/mL), followed by H. fomes (74.1 µg/mL) and C. decandra (116.7 µg/mL); however, lowest cytotoxicity was shown for A. corniculatum, A. officinalis, and S. caseolaris extracts. Mice treated with A. corniculatum extract did not experience any acute toxicity. These results demonstrate that S. caseolaris and A. corniculatum fruits have potential to treat diarrhea and may act as useful nutraceuticals.

Antidiarrheal, Analgesic, and Anthelmintic Activities of Honeys in the Sundarbans Mangrove Forest, Bangladesh.

This study evaluated the antidiarrheal, analgesic, and anthelmintic activities of honey samples from the Sundarbans mangrove forest of Bangladesh. Composite raw honey (RH), and its diethyl ether (DEH), ethanol (ETH), methanol (MEH), and distilled water (DWH) fractions were investigated. RH and its fractions strongly inhibited castor oil-induced diarrheal episodes in mice at a concentration of 250 mg/kg body weight (b.w.) (P<0.05). At this concentration, RH, DEH, ETH, MEH, and DWH showed inhibitory activity on diarrheal episodes at 43.8, 47.4, 29.8, 12.3, and 38.5%, respectively, whereas for the inhibitory activity for the positive control (PC, 3 mg loperamide/kg b.w.) was 47.4%. Similarly, DEH (250 mg/kg b.w.) showed strongest inhibition (63.5%) of acetic acid-induced writhing in mice, followed by RH (55.7%), ETH (46.2%), MEH (37.6%), and DWH (32.9%). In a hot plate test, mice treated with DEH at a concentration 250 mg/kg b.w. showed the greatest increase in response time, followed by treatment with RH. RH was further used in an anthelmintic test, where it showed a strong dose-dependent reduction in both the paralysis time and the time until death of the parasite, Paramphistomum cervi. Honeys in the Sundarbans could therefore be of great use as nutraceuticals.

Physicochemical and Antioxidant Properties of Honeys from the Sundarbans Mangrove Forest of Bangladesh.

This study evaluated the physicochemical, nutritional, antioxidant, and phenolic properties of ten honey samples from the Sundarbans mangrove forest, Bangladesh. The average pH, electrical conductivity, total dissolved solid, ash, moisture, hydroxymethyl furfural, titrable acidity, and absorbance were 4.3, 0.38 mS/cm, 187.5 ppm, 0.14%, 17.88%, 4.4 mg/kg, 37.7 meq/kg, and 483 mAU, respectively. In the honeys, the average contents of Ca, Cu, Fe, K, Mg, Mn, and Na were 95.5, 0.19, 6.4, 302, 39.9, 3.4, and 597 ppm, respectively, whereas Cd, Cr, Pb, and Ni were not found. The average contents of total sugar, protein, lipid, vitamin C, polyphenols, flavonoids, and anthocyanins in the honeys were 69.3%, 0.8%, 0.29%, 107.3 mg/kg, 757.2 mg gallic acid equivalent/kg, 43.1 mg chatechin equivalent/kg, and 5.4 mg/kg, respectively. The honeys had strong 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, reducing power and total antioxidant capacity. High-performance liquid chromatography analysis of the honey fractions revealed the quantification of six polyphenols namely, (+)-catechin, (-)-epicatechin, p-caumeric acid, syringic acid, trans-cinnamic acid, and vanillic acid at 194.98, 330.34, 74.64, 218.97, 49.55, and 118.84 mg/kg, respectively. Therefore, the honeys in the Sundarbans are of excellent quality and a prospective source of polyphenols, and antioxidants.

Antibacterial, Anti-Diarrhoeal, Analgesic, Cytotoxic Activities, and GC-MS Profiling of Sonneratia apetala (Buch.-Ham.) Seed.

Fruits of Sonneratia apetala (Buch.-Ham.), (English: mangrove apple, Bengali: keora) both seeds and pericarps, are largely consumed as food besides their enormous medicinal application. The fruit seeds have high content of nutrients and bioactive components. The seeds powder of S. apetala was successively fractionated using n-hexane, diethyl ether, chloroform, ethyl acetate, and methanol. The fractions were used to evaluate antibacterial, anti-diarrhoeal, analgesic, and cytotoxic activities. Methanol fraction of seeds (MeS) stronly inhibited Escherichia coli strains, Salmonella Paratyphi A, Salmonella Typhi, Shigella dysenteriae, and Staphylococcus aureus except Vibrio cholerae at 500 µg/disc. All the fractions strongly inhibited castor oil induced diarrhoeal episodes and onset time in mice at 500 mg extract/kg body weight (P<0.001). At the same concentration, MeS had the strongest inhibitory activity on diarrhoeal episodes, whereas the n-hexane fraction (HS) significantly (P<0.05) prolonged diarrhoeal onset time as compared to positive control. Similarly, HS (P<0.005) inhibited acetic acid induced writhing in mice at 500 mg extract/kg, more than any other fraction. HS and diethyl ether fractions of seed strongly increased reaction time of mice in hot plate test at 500 mg extract/kg. All the fractions showed strong cytotoxic effects in brine shrimp lethality tests. Gas chromatography-mass spectrometry analysis of HS led to the identification of 23 compounds. Linoleic acid (29.9%), palmitic acid (23.2%), ascorbyl palmitate (21.2%), and stearic acid (10.5%) were the major compounds in HS. These results suggest that seeds of S. apetala could be of great use as nutraceuticals.

Effects of beer and hop on ionotropic gamma-aminobutyric acid receptors.

Beer induced the response of the ionotropic gamma-aminobutyric acid receptors (GABA(A) receptors) expressed in Xenopus oocytes, indicating the presence of gamma-aminobutyric acid (GABA)-like activity. Furthermore, the pentane extract of the beer, hop (Humulus lupulus L.) oil, and myrcenol potentiated the GABA(A) receptor response elicited by GABA. The GABA(A) receptor responses were also potentiated by the addition of aliphatic esters, most of which are reported to be present in beer flavor. Aliphatic esters showed the tendency to decrease in the potentiation of the GABA(A) receptor response with an increase in their carbon chain length. When myrcenol was injected to mice prior to intraperitoneal administration of pentobarbital, the pentobarbital-induced sleeping time of mice increased additionally. Therefore, the beer contained not only GABA-like activity but also the modulator(s) of the GABA(A) receptor response.

Effect of 3-O-octanoyl-(+)-catechin on the responses of GABA(A) receptors and Na+/glucose cotransporters expressed in xenopus oocytes and on the oocyte membrane potential.

Recently, 3-O-octanoyl-(+)-catechin (OC) was synthesized from (+)-catechin (C) by incorporation of an octanoyl chain into C in the light of (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), which are the major polyphenols found in green tea and have strong physiological activities. OC was found to inhibit the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA(A) receptors) and Na+/glucose cotransporters expressed in Xenopus oocytes in a noncompetitive manner more strongly than does C. OC also induced a nonspecific membrane current and decreased the membrane potential of the oocyte, and thus death of the oocyte occurred even at lower concentrations than that induced by C or EGCg. Although EGCg produced H2O2 in aqueous solution, OC did not. This newly synthesized catechin derivative OC possibly binds to the lipid membrane more strongly than does C, Ecg, or EGCg and as a result perturbs the membrane structure.

Effects of tea components on the response of GABA(A) receptors expressed in Xenopus Oocytes.

To study the effects of tea components on ionotropic gamma-aminobutyric acid (GABA) receptor response, ionotropic GABA receptors (GABA(A) receptors) were expressed in Xenopus oocytes by injecting cRNAs synthesized from cloned cDNAs of the alpha(1) and beta(1) subunits of the bovine receptors, and their electrical responses were measured by a voltage clamping method. Extracts of green tea, black tea, and oolong tea in an aqueous solution induced the GABA-elicited response, which showed that these teas contain GABA, whereas coffee does not. Caffeine weakly inhibited the response in a competitive manner (K(i) = 15 mM), and (+)-catechin inhibited it in a noncompetitive one (K(i) = 1.7 mM). Especially, two catechin derivatives, (-)-epicatechin gallate and (-)-epigallocatechin gallate, inhibited the response strongly. Alcohols such as leaf alcohol or linalool potentiated the response, possibly because their binding to the potentiation site enhances the GABA-binding affinity to GABA(A) receptors when they bind. Extracts of green tea made with ethyl ether, which must contain lipophilic components of green tea, inhibited the response elicited by GABA, possibly because the amounts of caffeine and catechin derivatives were much larger than fragrant alcohols in such extracts of tea.

Effects of coffee components on the response of GABA(A) receptors expressed in Xenopus oocytes.

The effects of both coffee components and coffee extract on the electrical responses of GABA(A) receptors expressed in Xenopus oocytes were studied by injecting cRNAs of the alpha(1) and beta(1) subunits of the bovine receptors. The aqueous extract of coffee dose-dependently inhibited the GABA-elicited responses, whereas the lipophilic extract of coffee by diethyl ether slightly potentiated it at low doses (0.1-0.4 microL/mL) but showed inhibition at high doses (0.5-0.8 microL/mL). Theophylline inhibited the response in a noncompetitive mechanism (K(i) = 0.55 mM), whereas theobromine and trigonelline hydrochloride inhibited it in a competitive manner, K(i) = 3.8 and 13 mM, respectively. Benzothiazole, catechol, 2,4-dimethylstyrene, guaiacol, 1-octen-3-ol, sotolone, and 2,3,5-trimethylphenol potentiated the responses significantly. Potentiation elicited by guaiacol and sotolone was independent of GABA concentrations, whereas that by 1-octen-3-ol was dependent. When 1-octen-3-ol (100 mg/kg) was orally administered to mice prior to intraperitoneal administration of pentobarbital, the sleeping time of mice induced by pentobarbital increased significantly.

Aging of whiskey increases the potentiation of GABA(A) receptor response.

It is known that the target of most mood-defining compounds such as ethanol is an ionotropic gamma-aminobutyric acid receptor (GABA(A) receptor). The potentiation of the response of these inhibitory neurotransmitter receptors induces anxiolytic, sedative, and anesthetic activities in the human brain. Because both extracts of whiskey by pentane and fragrant components in whiskey potentiate the GABA(A) receptor-mediated response, GABA(A) receptors were expressed in Xenopus oocyte by injecting cRNAs prepared from the cloned cDNA for the alpha(1) and beta(1) subunits of the bovine receptors in order to study the effects of whiskey itself on the GABA(A) receptor-mediated response. Whiskey itself also potentiated the electrical responses of GABA(A) receptors generally more than ethanol at the same concentration as that of the whiskey. The potentiation of the GABA(A) receptor-mediated response increased with the aging period of the whiskey. Inhalation of whiskey to mice increased the sleeping time induced by pentobarbital more than that of the same concentration of ethanol as the whiskey. These results suggest that not only ethanol but also minor components in whiskey play an important role in the potentiation of GABA(A) receptor-mediated response and possibly the sedative effect of whiskey. Although the minor components are present in extremely small quantities compared with ethanol in alcoholic beverages, they may modulate the mood or consciousness of humans through the potentiation of the GABA(A) receptor response after absorption into the brain, because these hydrophobic compounds are easily absorbed into the brain across the blood-brain barrier and are several thousands times as potent as ethanol in the potentiation of the GABA(A) receptor-mediated response.

Potentiation of the ionotropic GABA receptor response by whiskey fragrance.

It is well-known that the target of most mood-defining compounds is an ionotropic gamma-aminobutyric acid receptor (GABA(A) receptor). The potentiation of the response of these inhibitory neurotransmitter receptors induces anxiolytic, sedative, and anesthetic activity in the human brain. To study the effects of whiskey fragrance on the GABA(A) receptor-mediated response, GABA(A) receptors were expressed in Xenopus oocyte by injecting rat whole brain mRNA or cRNA prepared from the cloned cDNA for the alpha(1) and beta(1) subunits of the bovine receptors. Most whiskey components such as phenol, ethoxy, and lactone derivatives potentiated the electrical responses of GABA(A) receptors, especially ethyl phenylpropanoate (EPP), which strongly potentiated the response. When this compound was applied to mice through respiration, the convulsions induced by pentetrazole were delayed, suggesting that EPP was absorbed by the brain, where it could potentiate the GABA(A) receptor responses. The extract of other alcoholic drinks such as wine, sake, brandy, and shochu also potentiated the responses to varying degrees. Although these fragrant components are present in alcoholic drinks at low concentrations (extremely small quantities compared with ethanol), they may also modulate the mood or consciousness of the human through the potentiation of the GABA(A) receptor response after absorption into the brain, because these hydrophobic fragrant compounds are easily absorbed into the brain through the blood-brain barrier and are several thousands times as potent as ethanol in the potentiation of the GABA(A) receptor-mediated response.

Polyphenol-induced inhibition of the response of na(+)/glucose cotransporter expressed in Xenopus oocytes.

To study the effects of polyphenols on the Na(+)/glucose cotransporter (SGLT1) response, SGLT1 was expressed in Xenopus oocytes by injecting cRNA synthesized from the cloned cDNA of the small intestine cotransporter of rats, and the electrical response elicited by glucose or galactose was measured by a voltage clamping method. Most phenol derivatives had no effect on the response. However, the polyphenols (+)-catechin, (-)-epicatechin gallate (ECg), and (-)-epigallocatechin gallate (EGCg), which are components of green tea, caused an inhibition of the response, which was almost independent of glucose concentration. The inhibition constants were estimated to be 2.3 mM for (+)-catechin and 0.45 mM for both ECg and EGCg, assuming the noncompetitive inhibition mechanism. Saponin prepared from tea seeds also inhibited the response significantly. Tannic acid and aqueous extracts of teas induced nonspecific electrical responses in both cRNA-injected and noninjected oocytes at lower concentrations than those that caused an inhibition of the SGLT1 response when their dose-dependent effects were examined. These results are possibly helpful in the development of a dietary supplement for diabetic patients.

Fragrances in oolong tea that enhance the response of GABAA receptors.

We electrophysiologically investigated the effect of some fragrant compounds in oolong tea on the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABAA receptors) which were expressed in Xenopus oocytes. Of the tested fragrances in oolong tea, cis-jasmone, jasmine lactone, linalool oxide and methyl jasmonate significantly potentiated the response. Among these, cis-jasmone and methyl jasmonate potently potentiated the response, having a respective dissociation constant of the compound (Kp) and maximum potentiation (Vm) of 0.49 mM and 322% for cis-jasmone, and 0.84 mM and 450% for methyl jasmonate. Inhalation of 0.1% cis-jasmone or methyl jasmonate significantly increased the sleeping time of mice induced by pentobarbital, suggesting that these fragrant compounds were absorbed by the brain and thereby potentiated the GABAA receptor response. Both of these compounds may therefore have a tranquillizing effect on the brain.