Isolation of the bioactive compound(s) with antinociceptive activity in mice, from the fish Haruan, Channa striata
p. 419-421
Texte intégral
2Note portant sur l'auteur3
3Note portant sur l'auteur4
4Note portant sur l'auteur5
5Note portant sur l'auteur6
Introduction
6A freshwater fish species haruan, Channa striata (Bloch 1793; syn. Channa striatus), is carnivorous and air breathing. The fish is indigenous to Malaysia and extracts are commonly used to induce postoperative wound healing, especially women following caesarean births. Haruan-based cream is also effective for exfoliation in dermatitis, such as sclerosis, psoriasis, eczema and ichthyosis. The product was awarded a bronze medal at the Annual MINDEX/INNOTEX meeting Kuala Lumpur in 1996. Furthermore, the antinociceptive activity of Haruan extracts in mice was recognised by the Society of Anaesthesiologists as The Most Original Paper Commendation Award, at the Annual Scientific Meeting in Singapore on the 1 7th April 1997. We have recently demonstrated that the extract provided 100 % inhibition of peritoneal pain receptors and enhanced morphine responses (Mat Jais et al., 1997).
7The Haruan fish contains all the essential amino and fatty acids required for wound healing (Mat Jais et al., 1994; 1998). We have previously shown that the bioactive compound in haruan extract is stable to pH 6 to 8, not destroyed by high temperature (100°C) and is not digested by the enzymes a-amylase, protease and lipase (Dambisya et al., 1999).
Materials and methods
8A Fresh midline fillet of haruan, C. striata, Figure 1, was homogenised in chloroform:methanol 2:1 v/v and the aqueous portion retained. A 2 ml aliquot of the aqueous was filtered through either 5,000, 10,000 and 30,000 dalton (Nominal Molecular Weight Limit) Millipore Ultrafree-CL low binding cellulose filter and centrifuged for 10 min at 5,000 rpm. A Series of 0, 25, 50 and 100 % dilutions of these filtered solutions were prepared in distilled water. Subsequently, 1 ml of the 5,000 and 10,000 dalton filtered solutions were purified by preparative HPLC using a LiChrospher 100 RP-18 column (5 mm, 125 mm x 4 mm, Hewlett Packard, Germany) at 1 ml/min and detected by UV at 205 nm. The mobile phase used 5 % CH3OH/H2O for 20 min, followed by gradient elution to 100 % CH3OH.
9The fraction collected between -3 minutes was dried under vacuum yielding 30 mg of extract, reconstitute in distilled water to a 0.0005, 0.005, 0.05 and 0.5 mg/ml in distilled water and then used in abdominal constriction tests in mice according to the method described (Mat Jais et al., 1997). Acetic acid (0.6 %) used to induce pain in mice peritoneal cavity, was administered by an intra peritoneal injection in a volume of 10 ml/kg 30 after the subcutaneous administration of 0.3 ml of saline control and the Haruan extracts. The activity was calculated as the percentage inhibition of abdominal constrictions or writhing (Mat Jais et al., 1997).
Results and discussion
10Each of the 25,50 and 100 % of the 5,000, 10,000 and 30,000 dalton filtered samples (see Figure 2 for the 30,000 dalton samples), as well as the HPLC fraction (Rt = 1 - 3 min) produced inhibition in constriction (Table 1). These activities were dose dependent and significantly different at P < 0.01. The lower concentrations had produced somewhat inconsistent activities (Figure 3), except the 5,000 dalton sample where the activity was clearly a dose dependent and significantly different at P < 0.01.
11We have previously reported that the bioactive compound(s) is thought to be a polar macromolecule (Mat Jais et al., 1994). The present findings confirm the molecule is more than likely less than 5,000 daltons. It remains a possibility that the compound(s) is a protein, perhaps a short peptide. It is unlikely however, to be a lipid such as a fatty acid given that fact that it is present in the aqueous fraction of the extraction.
12To date the identificaty of the bioactive compound(s) responsible for the antinociceptive activity remains to be determined and further analyses are currently in progress.
Table I. Antinociceptive activity of Haruan, Channa striata, HPLC purified filtered extracts at < 5,000 daltons
* Significant at P< 0.001 when compared to the control group
Figure 1. Picture of Haruan, Channa striata
Figure 2. Antinociceptive activity of Haruan, Channa striata, non-filtered and filtered extracts at < 30,000 daltons
Figure 3. Antinociceptive activity of Haruan, Channa striata, non-filtered and filtered extracts at <30,000, <10,000 and < 5,000 daltons
Figure 4. The analytical HPLC trace ofHaruan, Channa striatus, of filtered extracts at 5,000 daltons
Bibliographie
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Format
Reference
DAMBISYA Y.M., LEE T.L., SATHIVULU V., MAT JAIS A.M. (1999) The influence of temperature, pH and naloxone on the antinociceptive activity of Channa striata (haruan) extracts in mice, J Ethnopharmacol, 66, 2, 18-186.
10.1016/0306-3623(94)90101-5 :MAT JAIS A.M., MCCULLOCH R., CROFT K.D. (1994) Fatty acid and amino acid composition in haruan as a potential role for wound healing, Gen Pharmacol, 25, 5, 947-950.
MAT JAIS A.M., DAMBISYA Y.M., LEE T.L. (1997) Anti-nociceptive activity of Channa striata (haruan) extracts in mice, J Ethnopharmacol, 57, 2, 125-130.
MAT JAIS A.M., MATORI M.F., KITTAKOOP P., SUWANBORIRUX K. (1998) Fatty acid compositions in mucus and roe of haruan, Channa striata, for wound healing, Gen Pharmacol, 30, 4, 561-563.
Notes de fin
1 Department of Biomedical Sciences, University Putra Malaysia 43400 UPM Serdang Selangor (Malaysia) Email: mailto:abmanan@medic.upm.edu.my
2 Corresponding Author
3 Department of Biomedical Sciences, University Putra Malaysia 43400 UPM Serdang Selangor (Malaysia) Email: mailto:abmanan@medic.upm.edu.my
4 University Department of Medicine, University of Western Australia Perth, Western Australia (Australia) 6001
5 University Department of Medicine, University of Western Australia Perth, Western Australia (Australia) 6001
6 University Department of Medicine, University of Western Australia Perth, Western Australia (Australia) 6001
Auteurs
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