Abdul Manan Mat Jais

Fatty acid and amino acid composition in haruan as a potential role in wound healing

1. Two species of snakehead fish are available in Sabah, i.e. Channa striatus and Channa melanosoma, and are commonly known as haruan. Haruan is consumed by many Malaysians to induce healing after a clinical operations. However, there is no scientific evidence as yet to substantiate the claim, and so it was decided to analyse the biochemical composition in haruan to determine which compounds may have a possible role or potential in wound healing. 2. Samples (midline fillet) of both species were extracted separately in hexane for the qualitative analysis of fatty acids by a gas chromatography, Hewlett-Packard 5890A, using a 10 meter superox 11 column (Alltech) at temperature between 190 and 245 degrees C. Peak areas were calculated automatically using Hewlett-Packard 3393A computing integrator. Subsequently, the amino acid composition was analysed using a precolumn derivatization reverse phase HPLC waters PICO-TAG system. 3. Haruan is found to contain unusually high arachidonic acid (AA) but almost no eicosapentaenoic acid (EPA). AA which is a precursor of prostaglandin may initiate blood clotting and be responsible for growth. Haruan also contains all the essential amino acids for wound healing, particularly glycine which is the most important component of human skin collagen. Therefore, haruan contained all the basic biochemical requirements for wound healing.

Antinociceptive activity of Channa striatus (haruan) extracts in mice

Haruan, Channa striatus, is a snakehead fish consumed in many parts of the southeast Asian region. It is believed to promote wound healing, as well as reduce post-operative pain. In an attempt to establish the scientific basis for the alleged pain-relieving benefits of this fish, we studied the antinociceptive effects of whole fillet and mucus extracts from haruan in the mouse using the abdominal constriction and tail flick tests. In the abdominal constriction test, the 30 min fillet extract exhibited concentration-dependent inhibition of the writhing response in the 10-50% concentration range, with 20% as the IC50 value. This activity was not dependent on the duration of extraction, with no significant differences among the extracts obtained at durations of 10, 20, 30, 60, 90 and 120 min (range between 45-54% inhibition at 20% concentration). The mucus extract also showed concentration-dependent inhibition of the abdominal constriction response-at the highest concentration used the average inhibition was 68.9%, while IC50 value was 25%. Neither the fillet extract (30 min, 20%) nor the mucus extract (25%) had any demonstrable effect on the tail flick latency on their own, but significantly enhanced the antinociceptive activity of morphine in this assay. Similarly, low concentrations of the mucus and fillet extract enhanced the effects of morphine in the abdominal constriction test. Collectively, these results suggest a scientific basis for the folklore practice of eating haruan fish in the post-operative period for pain relief: Haruan extracts have antinociceptive activity and enhance the activity of other antinociceptive agents.

Influence of temperature, pH and naloxone on the antinociceptive activity of Channa striatus (haruan) extracts in mice

We have previously shown that extracts from the snakehead fish, Channa striatus (haruan), have antinociceptive activity on their own, and enhance morphine antinociception. In furtherance of that study, we investigated the effects of temperature, pH and naloxone on the antinociceptive effects of haruan extracts in the mouse abdominal constriction test. The effects of temperature were only studied on mucus extracts since the fillet extract is prepared at high temperature under pressure. There was a general loss of inhibitory activity from 88.4% at 0 degrees C to 61.1% at 80-100 degrees C, though this was only significant at 80 degrees C. The effects of pH were studied on both extracts. Lowering the mucus extract pH to 6.0 (unadjusted pH 7.4) did not significantly affect its activity, while raising it to 8.0 attenuated its activity. Similarly, pH changes had no effect on the fillet extract except at pH 8.0 which produced apparently lower antinociception. Pretreatment with naloxone (4 mg/kg) had no significant effect on the inhibitory effects of either extract in the abdominal constriction test. These results suggest that the active ingredient(s) of these extracts is (are) relatively stable over a wide range of temperature and between pH 6.0 and 8.0. That would be consistent with the possibility of a glycoprotein, polypeptide or polysaccharide.

The antinociceptive action of aqueous extract from Muntingia calabura leaves: the role of opioid receptors.

Objective: The present study was carried out to investigate the antinociceptive activity of the aqueous extract of Muntingia calabura (MCAE) leaves and to determine the effect of temperature and the involvement of the opioid receptor on the said activity using the abdominal constriction test (ACT) and hot-plate test (HPT) in mice. Materials and Methods: The extract was prepared by soaking the dried powdered leaves of M. calabura in distilled water (dH2O) overnight, and the supernatant obtained was considered as a stock solution with 100% concentration. The stock solution was diluted to 1, 5, 10, 50 and 100% and used to determine the antinociceptive activity of MCAE. A further experiment was done with 50% concentration to determine the effect of temperature and naloxone involvement of the opioid receptor system in MCAE antinociceptive activity. Results: At the various concentrations MCAE showed significant antinociceptive activity in both tests. However, the concentration-dependent activity was observed only in the ACT but not in the HPT. The 50% concentration of MCAEs were also stable against the effect of various temperatures as indicated by the presence of activity in both tests. The temperatures (40, 60 and 100°C) also showed an enhanced extract activity only in the HPT. Pre-treatment with naloxone (2 and 10 mg/kg) blocked the extract activity in both tests, indicating the involvement of the opioid receptor system in MCAE antinociceptive activity. Conclusion: Our data indicate that M. calabura leaves possess antinociceptive activity against chemically and thermally induced noxious stimuli. The bioactive compound(s) responsible for its antinociceptive activity is/are heat-stable and work partly via the opioid receptor system.

Fatty acid compositions in mucus and roe of haruan, Channa striatus, for wound healing

1. Fatty acid profiles in the external mucus extract and roe of Channa striatus were determined using gas chromatography (GC). 2. The mucus samples were collected by inducing hypothermic stress (-20 degrees C) for about 1 hr, and the roe were collected from gravid females at night soon after they liberated their eggs in a spawning program. 3. All mucus and roe samples were freeze-dried, except a part of roe which was not. 4. The mucus extract contained unsaturated fatty acid (oleic acid, C18:1 and linoleic acid, C18:2) as a major component, 21.25% and 22.47% of total lipid. 5. For the freeze- and nonfreeze-dried roe, the major components of fatty acid were somewhat similar to the mucus but with higher percentages: 58.56%, 26.08% and 45.76%, 20.94%. Interestingly, the nonfreeze-dried roe contained a large proportion of arachidic acid, C20:0 (22.16%), which was totally absent in the freeze-dried roe samples. 6. This profiling of the fatty acid mucus extract and roe is useful in strengthening the earlier claims that haruan possesses a potential remedy for wound healing (Mat Jais et al., 1994). Therefore, we are discussing the possibility of getting an optimum amount of the essential fatty acid for wound healing from various other parts of the fish without sacrificing the fish.

The antinociceptive activity of Muntingia calabura aqueous extract and the involvement of L-arginine/nitric oxide/cyclic guanosine monophosphate pathway in its observed activity in mice.

The present study was carried out to investigate on the possible involvement of l‐arginine/nitric oxide/cyclic guanosine monophosphate (l‐arginine/NO/cGMP) pathway in the aqueous extract of Muntingia calabura (AEMC) leaves antinociception in mice assessed by abdominal constriction test. The AEMC, obtained by soaking the dried leaves in distilled water (DH2O) (1 : 2; w/v) for 24 h, was prepared in concentrations of 10%, 50% and 100% that were approximately equivalent to doses of 27, 135 and 270 mg/kg, and administered subcutaneously (s.c.) 5 min after pre‐treatment (s.c.) of mice with DH2O, l‐arginine (20 mg/kg), NG‐monomethyl‐l‐arginine acetate (l‐NMMA; 20 mg/kg), NG‐nitro‐l‐arginine methyl esters (l‐NAME; 20 mg/kg), methylene blue (MB) (20 mg/kg), respectively. The AEMC was found to exhibit a concentration‐dependent antinociception after pre‐challenge with DH2O. Interestingly, pre‐treatment with l‐arginine was found to block significantly (P < 0.05) the AEMC antinociception but only at the highest concentration (100%) of AEMC used. On the other hand, pre‐treatment with l‐NAME was found to significantly (P < 0.05) enhance the low concentration but inhibit the high concentration AEMC antinociception. MB was found to significantly (P < 0.05) enhance AEMC antinociception at all concentrations used. Except for the higher concentration of AEMC used, co‐treatment with l‐NAME was found to insignificantly and significantly (P < 0.05) reverse the l‐arginine effect when given alone or with low concentration AEMC, respectively. In addition, co‐treatment with MB significantly (P < 0.05) reversed the l‐arginine effect when given alone or with 10% concentration AEMC but failed to affect the activity of the rest of concentrations used. As a conclusion, this study has demonstrated the involvement of l‐arginine/NO/cGMP pathway in AEMC antinociception.

Prostaglandin Analogous and Antioxidant Activity Mediated Gastroprotective Action of Tabernaemontana divaricata (L.) R. Br. Flower Methanolic Extract against Chemically Induced Gastric Ulcers in Rats

The present study was conducted to evaluate the antiulcerogenic effect and recognize the basic mechanism of action of Tabernaemontana divaricata (L.) R. Br. flowers. T. divaricata flower methanolic extract (TDFME) was screened for antiulcer activity versus aspirin and ethanol induced gastric ulcers at three doses—125, 250, and 500 mg/kg—orally using misoprostol as a standard. Besides histopathological examination, seven parameters, that is, ulcer index, total protein, nonprotein sulphhydryls, mucin, catalase, malondialdehyde, and superoxide dismutase levels, were estimated. In addition to HPLC profiling, GC-MS analysis and electrospray ionization—high resolution mass spectral (ESI-HRMS) analysis of crude TDFME were carried out in an attempt to identify known phytochemicals present in the extract on the basis of m/z value. The results revealed a significant increase in the levels of catalase, superoxide dismutase, mucin, and nonprotein sulphhydryls, while they revealed a reduction in ulcer index, the levels of total protein, and malondialdehyde. Histopathological observations also demonstrated the protective effect. Though all the doses of TDFME exhibited gastroprotective function, higher doses were found to be more effective. Mass spectral analysis gave a few characteristic m/z values suggesting the presence of a few known indole alkaloids, while HPLC profiling highlighted the complexity of the extract. TDFME was found to exhibit its gastroprotective effect through antioxidant mechanism and by enhancing the production of gastric mucous.

PCR-RFLP analysis of mitochondrial DNA cytochrome b gene among Haruan (Channa striatus) in Malaysia.

Haruan (Channa striatus) is in great demand in the Malaysian domestic fish market. In the present study, mtDNA cyt b was used to investigate genetic variation of C. striatus among populations in Peninsular Malaysia. The overall population of C. striatus demonstrated a high level of haplotype diversity (h) and a low-to-moderate level of nucleotide diversity (π). Analysis of molecular variance (AMOVA) results showed a significantly different genetic differentiation among 6 populations (F(ST)=0.37566, P=0.01). Gene flow (Nm) was high and ranged from 0.32469 to infinity (∞). No significant relationship between genetic distance and geographic distance was detected. A UPGMA tree based on the distance matrix of net interpopulation nucleotide divergence (d(A)) and haplotype network of mtDNA cyt b revealed that C. striatus is divided into 2 major clades. The neutrality and mismatch distribution tests for all populations suggested that C. striatus in the study areas had undergone population expansion. The estimated time of population expansion in the mtDNA cyt b of C. striatus populations occurred 0.72-6.19 million years ago. Genetic diversity of mtDNA cyt b and population structure among Haruan populations in Peninsular Malaysia will be useful in fisheries management for standardization for Good Agriculture Practices (GAP) in fish-farming technology, as well as providing the basis for Good Manufacturing Practices (GMP).

Salutary value of haruan, the striped snakehead Channa striatus – a review

Murrel namely Channa striatus or haruan contains all essential elements to maintain good health and to recover the lost energy after prolonged illness. The fatty acid composition (% of total fatty acid) indicated the abundant presence of C16:0 fatty acid as 30% and the other major fatty acids were C22:6 (15%), C20:4 (19%), C18:1 (12%) and C18:0 (15%). Haruan contains arachidonic acid (C20:4) as 19.0%, a precursor for prostaglandin and thromboxane biosyntheses. Both fatty and amino acids are important components for wound healing processes. Both the fillet and mucus extracts of haruan were found to exhibit a concentration dependent antinociceptive activity. In vitro antioxidant activity was higher in Channa roe protein hydrolysate than in Labeo roe protein hydrolysate in both DPPH radical scavenging and ferric reducing power. Protein content of roe concentrates (RPC) was found to be 90.2% (Channa) and 82.5% (Lates). Water absorption, oil absorption, foam capacity, stability and emulsifying capacity were found to be higher in Channa RPC than in Lates RPC. Characterization of protein hydrolysates from muscle and myofibrillar samples of haruan showed different kinetic and proteolytic activities. The skin extract of haruan influences the serotonergic receptor system thus they can function as an anti-depressant. Thus, haruan is the best example for food as medicine.

Gastroprotective Effect of Freeze Dried Stripped Snakehead Fish (Channa striata Bloch.) Aqueous Extract against Aspirin Induced Ulcerogenesis in Pylorus Ligated Rats

Channa striata (Bloch.) is a fresh water fish belonging to the family Channidae. The stripped snakehead fish possesses wide range of medicinal properties. In view of traditional use of C. striata for wound healing, the present study was undertaken to investigate the beneficial effects of orally administered freeze dried aqueous extract of Channa striata (AECS) in experimentally induced gastric ulcers in Wistar rats. Aspirin induced ulcerogenesis in pyloric ligation model was used for the assessment of antiulcer activity and Ranitidine (50 mg/kg) was employed as the standard drug. The various gastric parameters like volume of gastric juice, pH, free and total acidities, ulcer index, and levels of antioxidant enzymes like catalase, superoxide dismutase, and lipid peroxidation marker malondialdehyde were determined. AECS at concentrations of 40% and 50% w/v significantly decreased the volume of gastric juice and increased the levels of catalase while considerable decrease in free and total acidities and increase in superoxide dismutase were observed with the treatment of standard drug and AECS (50% w/v). All the test doses of AECS markedly decreased ulcer index and malondialdehyde compared to the standard drug whereas AECS 30% w/v did not alter volume of gastric juice, pH, free and total acidities, catalase, and superoxide dismutase. From these findings, it can be concluded that AECS is devoid of acid neutralizing effects at lower doses and possesses antisecretory and antiulcer activities and this could be related to its antioxidant mechanism.