Possible Antinociceptive Mechanism and Site of Activity of Haruan (Channa Striatus) Crude Aqueous Extract in Mice
The present study was carried out to determine the possible mechanism of antinociception and site of activity of the crude aqueous extract of Haruan (Channa striatus) (ASH) in mice using the abdominal constriction test. The ASH, obtained after chloroform:methanol (CM) (2:l; vlv) extraction (24 hr...
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| Format: | Thesis |
| Language: | English English |
| Published: |
2005
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| Online Access: | http://psasir.upm.edu.my/id/eprint/6324/1/FPSK%28P%29_2005_5.pdf |
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| Summary: | The present study was carried out to determine the possible mechanism of
antinociception and site of activity of the crude aqueous extract of Haruan (Channa
striatus) (ASH) in mice using the abdominal constriction test. The ASH, obtained after
chloroform:methanol (CM) (2:l; vlv) extraction (24 hrs) of the fresh Haruan fillet, was
evaporated to remove the methanol residue and used throughout the study. The first study
was carried out to ascertain the dry weight and antinociceptive profile of ASH. The
second study was carried out to determine the amino acids and fatty acids compositions,
as well as the polypeptide profile of ASH. The third study was carried out to determine
the actual onset and offset of ASH activity after its subcutaneous (SC) or intraperitoneal
(IP) administration at four different sets of time (0, 5, 30 and 60 min). The fourth and
fifth studies were carried out to determine the involvement of opioid and non-opioid
receptors, respectively, in the ASH antinociceptive activity. All of the antagonists of
opioidergic, muscarinic, nicotinic, a- and P-adrenergic, dopaminergic, serotonergic and
y-aminobutyric acid (GABA) receptors were administered (SC) 10 min prior to ASH
(SC) administration. The sixth study was carried out to determine the role of L argininelnitric oxide/cyclic 3'5'-guanosine monophosphate (L-arginine/NO/cGMP)
pathway in the ASH antinociceptive activity. The precursor (L-arginine) and inhibitor
@IG-nitro-L-arginine methyl esters (L-NAME)) for NO, as well as the inhibitor for cGMP
(methylene blue (MB), were administered (SC) 5 min before ASH administration (SC).
In all of the above-mentioned studies that involved the use of antinociceptive test, the
0.6% acetic acid-induced abdominal constriction test in mice was used as an assay to
evaluate the ASH antinociceptive activity. All data obtained were analysed using the
One-way Analysis of Variance (ANOVA) followed by the Tukey test with P<0.05 as the
limit of significance.
From the data obtained, the ASH, which exhibited significant (P<0.05) and
concentration/dosage-dependent antinociceptive activity, yielded 1.89g/10.0ml of white
coloured powder after subjection to the freeze-drying process. The ASH was also found
to contain all the important amino acids with major amino acids found are glycine
(35.77% -+ 0.58), alanine (10.19% + 1.27), lysine (9.44 + 0.56), aspartic acid (8.53 -+
1.15) and proline (6.86% -+ 0.78). Furthermore, the ASH was also found to contain high
composition of palmitic acid (C16:O) (35.93% * 0.63), oleic acid (C18: 1) (22.96% A
0.40), stearic acid (C18:O) (15.31% * 0.33), linoleic acid (C18:2) (1 1.45% * 0.3 1) and
arachidonic acid (C20:4) (7.44% rt 0.83). The ASH was also found to produce at least
four major fractions (at the retention times of 8.919, 9.841, 10.263 and 10.744), when
subjected to the high performance liquid chromatography (HPLC) process, that are
believed to be polypeptides. The onset time and the offset time of the ASH
antinociceptive activity, which are concentration-dependent and concentration independent, occurred between 0 to 5 min, and 60 min after its SC administration.
Interestingly, changing the route of administration from SC to IP caused significant
(Pc0.05) increase in the ASH antinociceptive activity with the concentration-independent
onset time of activity observed immediately after the ASH administration with no
apparent offset time. The activity was found to reach the maximum effect 30 min after
the ASH administration regardless of the route of administration used. Pretreatment with
naloxone at all dosages did not cause any significant changes in the ASH antinociceptive
activity indicating that the activity did not involve an opioid receptor mechanism, and
thus confirmed the report made by Dambisya et al. (1999). Re-treatment with various
types of non-opioid receptor antagonists demonstrated the involvement of at least four
=of receptors (muscarinic, GABA*, a-adrenergie d s e r o t o ~ g k ) in the
mechanism of ASH antinociceptive activity. Pre-treatment with atropine and bicuculine
almost completely blocked (P<0.05), while pre-treatment with phenoxybenzamine and
methysergide significantly (Pc0.05) reduced half of the ASH activity. The role of Larginine/
NO/cGMP pathway in ASH antinociceptive activity was also observed after
pretreatment of the ASH with L-arginine, L-NAME or MB, but not with D-arginine.
Pretreatment with L-arginine was found to significantly (PC0.05) reduce the ASH
antinociceptive activity, whereas pretreatment with L-NAME or MB were found to
enhance (Pc0.05) the activity. Based on the finding, low concentration of NO, limited by
the presence of higher concentration of ASH, and inhibition of cGMP system play
important role in ASH antinociceptive activity. However, the actual mechanism
underlying this phenomenon is yet to be fully understood.As a conclusion, we suggest that the ASH-produced antinociceptive activity could be due
to the presence of various types of amino acids and fatty acids, as well as four major
fractions, and involved activation of at least four types of the non-opioid receptors
(namely the muscarinic, GABA*, a-adrenergic and serotonergic) and the Larginine/
NO/cGMP pathway. |
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