Professional Documents
Culture Documents
net/publication/236247048
CITATIONS READS
20 184
5 authors, including:
Some of the authors of this publication are also working on these related projects:
Mechanism of analgesisc activities of fractions obtained from methanol leaf and root bark extract of Cissus cornifolia Planch. (Vitaceae) View project
All content following this page was uploaded by Mohammed Garba Magaji on 23 May 2014.
ABSTRACT
The methanolic leaf extract of Securinega virosa was evaluated for possible analgesic and anti-inflammatory activities in
rodents. Acetic acid induced writhing test in mice and formalin-induced pain in rats were used to study the analgesic
effect, while the effect of the extract on acute inflammation was investigated on carrageenan-induced paw oedema in
rats. The extract significantly (P < 0.01) inhibited acetic acid induced writhing in mice and significantly (P < 0.05)
attenuated the neurogenic phase of formalin-induced pain in rats at the highest dose tested. The extract also produced a
moderate anti-inflammatory activity which was found to be significant (P < 0.05) at all the doses tested. Preliminary
phytochemical screening of the extract revealed the presence of alkaloids, tannins. saponins, cardiac glycosides,
flavonoids and resins. The intraperitoneal median lethal dose (LD 50) of the extract in mice was found to be 1,265 mg/kg
suggesting that the extract may be relatively safe at the analgesic doses. The results obtained in this study lend credence
to the ethnomedical use of the plant in the management of pain and inflammatory conditions.Thus, supporting the
development of the biologically active substances as analgesics and anti-inflammatory agents.
47
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
to overcome these shortcomings has become a investigated, this present study was aimed at
major goal in pain research. In view of this investigating the analgesic and anti-
and on account of the ethnomedical claim of inflammatory activities of the methanolic leaf
the usefulness of the plant in the management extract of Securinega virosa in laboratory
of pain and inflammatory conditions, which to animals.
our knowledge has not been scientifically
48
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
The phytochemical screening of M. angolensis on the following scale: (0) rat walked or stood
was carried out on the methanolic root bark firmly on the injected paw; (1) the injected
extract using standard protocol (Trease and paw was favoured or partially elevated; (2) the
Evans, 1983). injected paw was clearly lifted off the floor;
(3) the rat licked, chewed or shook the
Acetic acid induced writhing test in mice injected paw. Anti-nociceptive effect was
Mice were divided into five groups each determined in two phases. The early phase
containing 6 mice. The control group received (phase 1) was recorded during the first
Normal saline (10ml/kg, i.p.). The test groups 5minutes, while the late phase (phase 2) was
were treated with 25, 50, 100 mg/kg/i.p. of recorded during the last 45 minutes with a 10
SVMLE while the fifth group received minutes lag period in-between both phases.
piroxicam at the dose of 10mg/ kg, i.p. After (Dubuisson and Dennis, 1977; Tjølsen et al.,
30 minutes of drug administration, the mice 1992).
were treated with 0.6% acetic acid at 10ml/kg
body weight, i.p. (Koster et al., 1959). Five Carrageenan-induced paw oedema
minutes after acetic acid injection, mice were The rats were divided into five groups each
placed in individual cage and the number of containing 5 rats. Acute inflammation was
abdominal contractions was counted for each induced by injecting 0.1 ml of (1%)
mouse for a period of 10 minutes after 5 carrageenan into plantar surface of rat hind
minutes latency, and the percentage inhibition paw (Winter et al., 1962). The methanolic leaf
of writhing was calculated. extract (25, 50 and 100 mg/kg), normal saline
(1ml/kg) and ketoprofen (10 mg/kg) as
Formalin test in rats reference agent were administered 30 min
The rats were divided into five groups each before carrageenan injection. The paw volume
containing 5 rats and were administered with was measured at 0, 1, 2, 3, 4 and 5h using a
either normal saline (1ml/kg, i.p.), methanolic vernier caliper to determine the diameter of
leaves extract (25, 50 and 100 mg/kg, i.p) or oedema. The difference between the readings
Morphine (4 mg/kg, s.c). Thirty minutes after at time 0 h and different time interval was
this treatment; 50 µl of a freshly prepared taken as the thickness of oedema.
2.5% solution of formalin was injected
subcutaneously under the plantar surface of Statistical analysis
the left hind paw of each rat. The rats were The results were expressed as Mean ± SEM
placed individually in an observation chamber and analysed using Student’s t-test. A P value
and monitored for one hour. The severity of < 0.05 was considered significant.
pain response was recorded for each rat based
49
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
Constituents Remark
Tannins +
Saponins +
Flavonoids +
Alkaloids +
Cardiac glycosides +
Cyanogenic glycosides +
Resins +
Steroid/Terpenoids +
Carbohydrates +
Anthraquinone -
Each value represents mean ± SEM. *P < 0.001 compared with control (Student’s t-test).
SVMLE= Methanolic leaf extract of S. virosa; n=6
Each value represents mean ± SEM. *P < 0.05 **P < 0.001 compared with control
(Student’s t-test) SVMLE= Methanolic leaf extract of S. virosa. n=5
50
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
1 2 3 4 5
N/Saline 0.19 ± 0.01 0.25 ± 0.03 0.23 ± 0.02 0.20 ± 0.03 0.20 ± 0.02
1ml/kg
SVMLE 0.16 ± 0.02 0.19 ± 0.02 0.16 ± 0.02* 0.13 ± 0.02* 0.12 ± 0.02*
25mg/kg
(15.8) (24.0) (30.4) (35.0) (40.0)
SVMLE 0.11 ± 0.01** 0.16 ± 0.03* 0.14 ± 0.02* 0.12± 0.02** 0.10 ± 0.02*
50mg/kg
(42.1) (36.0) (39.1) (40.0) (50.0)
SVMLE 0.10 ± 0.01** 0.15 ± 0.01* 0.14 ± 0.02* 0.09 ± 0.02** 0.08 ± 0.01**
100mg/kg
(47.4) (40.0) (39.1) (55.0) (60.0)
Ketoprofen 0.08 ± 0.02*** 0.07 ± 0.03*** 0.09 ± 0.03*** 0.10 ± 0.02** 0.09 ± 0.03**
10mg/kg
(57.9) (72.0) (91.3) (50.0) (55.0)
Each value represents mean ± SEM. *P < 0.05, **P <0.01 ***P <0.001, compared with control
values for corresponding hours. Figures in parentheses are percentage inhibition of inflammation.
SVMLE= Methanolic leaf extract of S. virosa. n= 5
51
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
the spinal level after peripheral inflammatory 3 hours (Brooks and Day, 1991). The extract
state (Diaz and Dickeson, 1997). The two moderately inhibited the carrageenan-induced
distinct phases in formalin test are due to inflammation in the 3rd hour. Analgesic and
direct effect of formalin on nociception and anti-inflammatory effects have been observed
due to inflammation with the release of in flavonoids as well as tannins (Ahmadiani et
serotonin, histamine, bradykinin and al., 2000). Flavonoids such as quercetin are
prostaglandins and at least to some degree, the known to be effective in acute inflammation
sensitization of central nociceptive neurons (Rajnarayana et al., 2001). There are also
(Dubuisson and Dennis, 1977; Huskaar and reports on the analgesics effects of alkaloids,
Hole, 1987; Tjøsen et al., 1992). Stimulation essential oils and saponins (Choi et al., 2005;
of opioid receptors has also been suggested as de Araujo et al., 2005; Reanmongkol et al.,
a possible mechanism of action against 2005).
neurogenic pain (Gaertner et al., 1999). The The analgesic and anti-inflammatory effect of
ability of the extract to inhibit the second the extract may therefore, be due to the
phase of formalin-induced pain suggests that it presence of flavonoids, tannins, alkaloid or
may possess central analgesic activity. The saponins. However, further studies are in
probable mechanism of action of carrageenan- progress in our laboratory to isolate the active
induced inflammation is bi-phasic, the first constituents responsible for the observed
phase is attributed to the release of histamine, effect, and to elucidate the possible
serotonin and kinins in the first hour; while mechanisms of action responsible for the
the second phase is attributed to the release of analgesic and anti-inflammatory activities of
prostaglandins and lysosome enzymes in 2 to the methanolic leaf extract.
REFERENCE
Choi J, Jung H, Lee K, Park H (2005). Antinociceptive
Ahmadiani, A., Hosseiny, J., Semnanian, S., Javan, M., and Antiinflammatory effects of saponin and sapogenin
Saeedi, F., Kamalinejad, M., and Saremi, S. (2000). obtained from the stem of Akebia quinata. Journal of
Antinociceptive and anti-inflammatory effects of Medicinal Food, 8 (1) 78-85.
Elaeagnus angustifolia fruit extract. Journal of
Ethnopharmacology, 72: 287–292. Collier, H. O. J., Dinnean, L. C., Johnson, C. A. and
Schenider, C. (1968); The abdominal constriction
Almeida, R.N., Navarro, D.S. and Barbosa-Filho, J.M. response and its suppression by analgesic drugs in the
(2001). Plants with central analgesic activity. mouse British Journal of Pharmacology, 32: 295-310
Phytomedicine, 8(4): 310-322.
Dalziel, J. M. (1936). The Useful Plants of West
Bentley, G.A., Newton, S.H. and Starr, J. (1981). Tropical Africa, Watmonghs, Idle, London, pp. 354-
Evidence for an action of morphine and enkephalins on 355.
sensory nerve endings in the mouse peritoneum.
British Journal of Pharmacology, 73: 325-332. de Araujo, P.F., Coelho-de-Souza, A.N., Morais, S.M.,
Ferreira, S.C. and Leal-Cardoso, J.H. (2005).
Brooks, P.M. and Day, R.O. (1991). Non steroidal anti- Antinociceptive effects of the essential oil of Alpinia
inflammatory drugs difference and similarities. New zerumbet on mice. Phytomedicine, 12: 482-686.
England Journal of Medicine, 324: 1716-1725.
52
Yerima et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 47 – 53
Derardt, R., Jongney, S., Delvalcee, F. and Falhout, M. dependent on the polymorphonuclear leucocytes.
(1980). Release of prostaglandins E and F in an Science, 225: 743-745.
algogenic reaction and its inhibition. European Journal
of Pharmacology, 51: 17-24. Lorke, D. (1983). A new approach to acute toxicity
testing. Archives of toxicology, 54:275-287.
Dhara, A.K., Suba, V., Sen, T., Pal, S. and Chaudhuri,
A.K. (2000). Preliminary studies on the anti- Naik, D.G., Mujumdar, A.M., Wagole, R.J., Kulkarni,
inflammatory and analgesic activity of methanolic D.K., and Kumbhojkar, M.S. (2000). Pharmacological
fraction of the root of Tragia involucrate. Journal of studies of Sterculia foetida leaves. Pharmacological
Ethnopharmacology, 72: 265-268. Biology, 1(38): 13-17.
Diaz, A. and Dickenson, A. H. (1997). Blockade of Neuwinger J.D. (translated from the German by Porter,
spinal N- and P-type, but not, L-type calcium channels A.) (1996). African Ethnobotany-Poisons and Drugs.
inhibits the excitability of rat dorsal horn neurones Chapman and Hall, Weinheim pp. 495-499.
produced by subcutaneous formalin inflammation.
Pain, 69: 93-100.
Rajnarayana, K., Sripal Reddy, M. and Chaluvadi,
M.R. (2001) Bioflavanoids Classification,
Dubuisson, D. and Dennis, S.R. (1977). The formalin Pharmacological, Biochemical effects and Therapeutic
test: A quantitative study of the analgesic effects of potential. Indian Journal of Pharmacology; 33: 2-16.
morphine, meperidine, and brain stem stimulation in
rats and cats. Pain, 4: 161-174. Reanmongkol, W., Subhadhirasakul, S., Thienmontree,
S., Thanyapanit, K., Kalnaowakul, J. and Sengsui, S.
Gaertner, M., Muller, L., Roos, J.F, Cani, G, Santos, (2005). Antinociceptive activity of the alkaloid extract
A.R., Niero, R., Calixto, J.B, Yunes, R.A., Delle from Kopsia macrophylla leaves in mice.
Monache, F. and Cechinel Filho, V. (1999) Analgesic Songklanakarin Journal of Science and Technolology,
triterpenes from Sebastiania schottiana roots. 27(Supplement 2): 509-516.
Phytomedicine, 6: 41-44.
Tjølsen, A., Berge, O., Hunskaar, S., Rosland, J.H. and
Huskaar, S. and Hole, K. (1987). The formalin test in Hole, K. (1992). The formalin test: An evaluation of the
mice: Dissociation between inflammatory and non method. Pain, 52: 5-17.
inflammatory pain. Pain, 30: 103-114.
Trease, G. E. and Evans, M.C. (1983) Textbook of
Koster, R., Anderson, M., De Beer, E.J. (1959). Acetic Pharmacology 12th ed., Balliere Tindall, London. pp.
acid for analgesic screening. Federation proceedings, 322-383.
18:412.
WHO/EDM/TRM/2000.1 General Guidelines for
Koyama, K., Imaizumi, T., Akiba, M., Kinoshita, K., Methodologies on Research and Evaluation of
Takahashi, K., Suzuki, A., Yano, S., Horie, S., Traditional Medicine. pp.1-20.
Watanabe, K. and Naoi, Y. (1997). Antinociceptive
components of Ganoderma lucidum. Planta medica, 63:
224-227. Winter, C. A., Riselay, E.A. and Nuss, G.W. (1962)
Carrageenan-induced oedema in the hind paw of the
rats as an assay for anti – inflammatory drugs. Proc.
Levini, J.D., Lau, W., Kwait, G. and Goetzl, E.J. soc. Exp. Biol. Med. 111: 544-547.
(1984). Leukotriene B4 produces hyperalgesia that is
53