Antinociceptive activity and effect of methanol extract of Salvia limbata on withdrawal syndrome in mice
←
→
Page content transcription
If your browser does not render page correctly, please read the page content below
European Review for Medical and Pharmacological Sciences 2012; 16: 38-42
Antinociceptive activity and effect of
methanol extract of Salvia limbata on
withdrawal syndrome in mice
Sh. ALEMY1, M. KARAMI2, E. HOSSINI3, M.A. EBRAHIMZADEH4, N. SHAHBI MAJD5
1
Department of Physiology and Pharmacology, Islamic Azad University Sciences and Research
Branc fars (Iran)
2
Department of Toxico-Pharmacology and Pharmaceutical Sciences Research Center, School of
Pharmacy, Mazandaran University of Medical Sciences, Sari (Iran)
3
Department of Physiology and Pharmacology, Islamic Azad University Sciences and Research
Branc fars (Iran)
4
Departmnet of Medicinal Chemistry, School of Pharmacy, Mazandaran University of Medical
Sciences, Sari (Iran)
5
Department of Physiology and Pharmacology, School of Medicine, Mazandaran University of
Medical Sciences, Sari (Iran)
Abstract. – Objectives: Salvia (S.) is an Conclusions: S. limbata extract produced sta-
impor tant genus consisting of about 900 tistically significant inhibition of pain and devel-
species in the Lamiaceae family. They are several opment of morphine dependence in mice.
reports that some Salvia spp. has effects on the
central nervous system (CNS). The present ex- Key Words:
periments were undertaken to study the protec-
tive effect of S. limbata on the development of Morphine dependence, Antinociceptive activity,
dependence to morphine in mice. Salvia limbata, Jumping, Hot plate.
Material and Methods: Antinociceptive ac-
tivity of aerial parts of S. limbata was investigat-
ed using the hot plate method. In addition, the
effect of its aerial parts on morphine depen-
dence was investigated in mice. After induction
of dependence by morphine, different concentra- Introduction
tions of plant aerial parts extract were injected
to treated groups. To assess morphine with- It is well clear that repeated use of opioid
drawal, mice were injected naloxone (5 mg/kg) drugs brings physical dependence and tolerance.
i.p. on the 5th day. After four consecutive days Based on evidence from neurochemical, neuro-
of morphine injection, withdrawal syndrome was
assessed by placing each mouse in a 30 cm
physiological and biochemical studies of opioid
high glass box and recording the frequency of dependence, a variety of agents and systems such
escape jumps for 60 minutes. as noradrenergic system1 , serotonergic2, adeno-
Results: Animal receiving acute treatment sine receptor agonists3, excitatory amino acid an-
with morphine displayed dependence. The ani- tagonists4,5, protein kinase C inhibitors6, gluco-
mals treated with different extract concentra- corticosteroids7, benzodiazepines8,9 and arachi-
tions could decrease frequency of escape jumps
donic acid10 can modulate the morphine with-
in number or decrease development of morphine
dependence. Addiction was observed following drawal syndrome.
naloxone administration. Methanol extract of S. Pain is still one of the main health problems of
limbata produced a statistically significant inhi- the world’s populations11. Many bioactive sub-
bition of pain induced by hot plate latency at stances are involved in the modulation of pain
500, 1000 and 1500 mg/kg i.p. A significant in- sensation 12 . Eclectic physicians relied upon
crease in pain threshold was observed after 30 herbal medicins and natural remedies to treat dis-
and 60 min (p < 0.001). The activity was compa-
rable to that of morphine (30 mg kg-1 i.p., p >
ease13.
0.05). The anti-nociceptive activity of S. limbata Salvia is an important genus consisting of about
increased until the 60th min (p < 0.05 compared 900 species in the Lamiaceae family14. They are
to morphine). several reports that some Salvia spp. has effects on
38 Corresponding Author: Mohamamd Karami, Ph.D; e-mail: toxkarami@gmail.comEffect of Salvia limbata on withdrawal syndrome
the central nervous system (CNS). S. haematodes freeze-dried.The extract was prepared in phos-
has CNS depressant, antinociceptive and anticon- phate buffer (pH 7.4) and tween 80 (4:1) for
vulsant activities15,16. The genus, Salvia (Labi- pharmacological studies.
atae), is generally known for its multiple pharma-
cological effects including analgesic and anti-in- Morphine Dependence
flammatory activities17,18, S. leriifolia has an effect Morphine was injected intraperitoneally (i.p.)
on morphine dependence19 and hypoglycaemic ef- to mice at doses of 50, 75,100 and 125 mg/kg
fects morphine dependency as well20. Anti-noci- three times daily (8:00 a.m., 12:00 and 16:00
ceptive and anti-inflammatory activities have also p.m., respectively) for 4 days. On day 5, a single
been reported for this species18. dose of morphine (50 mg/kg) was injected 2 h
Jumping is most suitable sign of measuring before naloxone treatment 21.
abstinence quantity as jumps are easily counted
and jumping rate increases when dependence in- Morphine Withdrawal
creases or dose of antagonist increased. Investi- Withdrawal signs were precipitated by injec-
gation on plant S. limbata revealed its beneficial tion of naloxone (5 g/kg, i.p.) 2 h after the final
effects to decrease dependence sign produced by administration of morphine. After the naloxone
morphine and increase in pain threshold after 60 challenge, mice were immediately placed in a
min, in comparison to the control. The present glass cylinder (30 cm high, 20 cm in diameter).
experiments were undertaken to study the protec- The number of jumping episodes was counted
tive effects of S. limbata extract on the develop- for 60 min after naloxone injection22.
ment of dependence to morphine in mice.
Extract Treatment
After induction of dependence by morphine,
mice were divided into 10 groups. Then the con-
Materials and Methods trol group was injected distilled water and differ-
ent concentrations of plant extract (100, 200,
Drugs 500, 1000, 1500 mg/kg) were injected to the oth-
Morphine sulphate was prepared from Daru er groups i.p. 1/5 h after the final dose of mor-
Pakhsh Co. (Iran) and naloxone hydrochloride phine.
was prepared from Tolid Daru Co. (Iran).
Antinociceptive Study
Animals The hot-plate test was assessed on male mice.
Male albino mice 25-30 g was obtained from a The temperature of the metal surface was main-
random bred colony, maintained on a special diet tained at 55± 0.2°C. Latency to a discomfort reac-
in the animal house of Sari University of Medical tion (licking paws or jumping) was determined be-
Sciences. The animals had free access to a stan- fore and after drug administration. The cut-off
dard commercial diet and water ad libitum and time was 55 s. Morphine was injected i.p.) to
were kept in rooms maintained at 25 ± 1°C with mice, as a single dose of 30 mg kg-1 (as a positive
a 12/12h light/dark cycle. control). Solvent was injected to the negative con-
trol group (10 mL kg -1 , i.p.). An aqueous
Plant Material methanolic extract of the aerial parts of S. limbata
Aerial parts (flowered browse) of S. limbata was given at doses of 500, 1000, 1500 mg kg-1 i.p.
were collected from Tehran and was identified to the animals. Antinociceptive activity was as-
and confirmed by Department of Pharmacognosy sessed by measuring the hot plate latency to heat,
(Dr. Gohari). A voucher specimen (No. 783) has as described by Eddy and Leimbach23.
been deposited in Tehran School of Pharmacy
Herbarium. Aerial parts were dried at room tem- Statistical Analysis
perature and coarsely ground before extraction. Statistical analysis was performed using the
One hundred grams of the powdered sample was SPSS software for Windows Ver.10, (SPSS Inc.,
extracted at room temperature by percolation Chicago, IL, USA). Data were analyzed by one-
with methanol/water (80:20, 400 ml × 3). The re- way analysis of variance (ANOVA) and present-
sulting extract was concentrated over a rotary ed as Mean ± SD. Student-Newman-Keuls test
vacuum evaporator, until a solid extract sample was used for statistical analysis and p < 0.05 was
was obtained. The resulting crude extract was considered to be significant.
39Sh. Alemy, M. Karami, E. Hossini, M.A. Ebrahimzadeh, N. Shahbi Majd
Jumping count/60 min
Control 100 mg/kg 200 mg/kg 500 mg/kg 1000 mg/kg 1500 mg/kg
Salvia limbata (mg/kg)
Figure 1. Relation between morphine withdrawal jumps and different concentration of plant methanol extract. Significant at
p < 0.0, each value represents Mean ± SD.
Results The extract reduced the jumping episodes dose-
dependently. The maximum effect was observed at
Animal receiving acute treatment with mor- a dose of 1 g kg-1. Results of the present study
phine displayed dependency. The animals treated showed that the aqueous methanol extract of the
with different Salvia limbata extract concentration aerial parts (flowered browse at 1000 mg kg-1) of
could decrease or increase frequency of escape S. limbata produced a statistically significant in-
jumps in number, following naloxone administra- crease in the pain threshold, after 30 min, in com-
tion. Recently, we have shown that the high inhibi- parison to the control (Figure 1). The effect or ac-
tion of morphine dependence in methanolic ex- tivity was rather low, however enough for treat-
tract of S. limbata can decrease development of ment and blocking the pain. This activity was
morphine dependence. However, mechanism of comparable to that of morphine (30 mg kg-1 i.p., p
plant action to S. limbata to inhibit or decrease ab- > 0.05). The anti-nociceptive activity of extract in-
stinence syndrome in dependent mice is unclear. creased until the 60th min.
Latency (sec)
Latency (sec)
Saline Morphine S. limbata Saline Morphine S. limbata
Figure 2. Anti-nociceptive activity of methanolic extract Figure 3. Antinociceptive activity of aqueous methanol
of S. limbata aerial parts after 30 min. Values are presented extract of S. limbata aerial parts after 60 min. Values are
as Mean ± SD (n = 7), ***p < 0.001 with respect to control presented as Mean ± SD (n = 7), ***p < 0.001 with respect
(ANOVA followed by Newman-Keuls multiple comparison to control (ANOVA followed by Newman-Keuls multiple
test). comparison test).
40Effect of Salvia limbata on withdrawal syndrome
Discussion leriifolia18 other genera33. This study and other
research on aerial parts of S. limbata also con-
The present results indicate that the macerated firm that Salvia genera are good candidates for
methanolic extract of S. limbata reduced the anti-inflammatory and analgesic uses. It is con-
withdrawal signs of morphine, dose-dependency. cluded that the methanol extract of S. limbata has
Adenosine A1 receptor agonists such as 2- a central (no spinal) antinociceptive effect and
chloroadenosine and R-phenylisopropyladeno- this may be mediated by opioid receptors.
sine suppressed the withdrawal syndrome of
morphine. Adenosine receptor antagonists such
as caffeine and theophylline increased the jump- ––––––––––––––––––––
ing episodes and blocked the effects of adenosine Acknowledgements
analogues3. S. miltorrhiza extract increased the This work was supported by a grant from the Research
ATP level in the brain24. As ATP is broken down Council of the Medical Sciences University of Mazan-
to adenosine25, it might be possible that the ex- daran/Iran.
tract decreased morphine dependence by an
adenosine mechanism. Further study is needed to
confirm this mechanism. Benzodiazepines, via
GABAA receptors had an inhibitory effect on the References
dependence to morphine 8,9. As some binding
1) AMBROSIO E, IGLESIAS V, GARCIA-LECUMBERRI C, OREN-
sites were found on the GABA/benzodiazepine SANZ L, ALGUACIL LF. Effect of yohimbine on the de-
receptor complex for some S. spp.26,27, there is al- velopment of morphine dependence in the rat:
so a possibility that S. limbata acts through this lack of involvement of cortical beta-adrenoceptor
complex to affect morphine dependency. modifications. Pharmacol Biochem Behav 1997;
The involvement of other mechanisms may al- 56: 487-491.
so be considered. S. miltorrhiza via danshen, a 2) H ARRIS GC, A STON JG. Augmented accumbal
constituent in the root, inhibited adenylate cy- serotonin levels decrease the preference for a
morphine associated environment during with-
clase activity in rat brain28. It also inhibited the drawal. Neuropsychopharmacol 2001; 24: 75-
phosphatidylinositol system in acute myocardial 85.
ischaemia29. Therefore, some Salvia genus may 3) MICHALSKA E, MALEC D. Agonist and antagonists of
potentially have inhibitory effects on the with- adenosine receptors and morphine withdrawal
drawal syndrome of morphine via these second syndrome in rats. Pol J Pharmacol 1993; 45: 1-9.
messenger systems, which have modulatory ef- 4) BELOZERTSEVA I, ZUARTAV E, BESPALOV A. Behavioral
fects on morphine dependency30,31. effect of MK-801 in morphine dependent and
In conclusion, the methanolic extract of S. lim- non-dependent mice. Life Sci 1996; 58: 55-61.
bata can suppress the morphine withdrawal syn- 5) GONZALEZ P, CABELLO P, GERMANY A, NORRIS B, CON-
TRERAS E. Decrease of tolerance to, and physical
drome. The results of this study are valuable as a dependence on morphine by glutamate receptor
step towards the search for different mechanism of antagonists. Eur J Pharmacol 1997; 332: 257-
actions, which may be involved in the inhibitory 262.
effect of the extract on morphine dependency. It is 6) TOKUYAMA S, FENG Y, WAKABAYASHI H, HO IK. Possible
difficult to speculate on the exact mechanism of ac- involvement of protein kinases in physical depen-
tion at this time. The present results indicate that dence on opioids: study using protein kinase C in-
hibitors, H7 and H8. Eur J Pharmacol 1995; 284,
the aqueous extract of S. limbata has central an- 101-107.
tinociceptive activity, because it showed a signifi-
7) CAPASSO A, PINTO A, SORRENTINO L, CIRINO G. Dex-
cant antinociceptive effect in the hot-plate test and amethasone inhibition of acute opioid physical
also its effect was inhibited by naloxone, a specific dependence in vitro is reverted by anti-lipocortin-
antagonist of opioid receptors. The inhibitory ef- 1 and mimicked by anti-type II extracellular PLA2
fect of naloxone on the antinociceptive activity of antibodies. Life Sci 1997; 61: 127-134.
extract suggests a morphine-like activity profile for 8) SUZUKI T, TSUDA M, NARITA M, FUNADA M, MIZOGUCHI
this plant. With regard to the LD50 value and in H, MISAWA M. Diazepam pretreatment suppresses
morphine withdrawal signs in the mouse. Life Sci
comparison with a toxicity classification32, the ex- 1996; 58: 349-357.
tract was of low toxicity.
9) PUNTILLO K, CASELLA V, REID M. Opioid and benzodi-
Antinociceptive and/or anti-inflammatory ac- azepine tolerance and dependence: application of
tivities have been reported for some Salvia gen- theory to critical care practice. Heart Lung 1997;
era such as S. hemaematodes15, S. aethiopis17, S. 26: 317-324.
41Sh. Alemy, M. Karami, E. Hossini, M.A. Ebrahimzadeh, N. Shahbi Majd
10) CAPASSO A, SORRENTINO L. Arachidonic acid and its and Hall, Harborne JB. 1998. 3rd ed. Chapman
metabolites are involved in the expression of mor- and Hall.
phine dependence in guinea-pig isolated ileum. 23) E DDY NB, L EIMBACH D. Synthetic analgesics. II.
Eur J Pharmacol 1997; 9: 330: 199-204. Dithienylbutenyl- and dithienylbutylamines. J
11) BASBAUM AI, FIELD HL. Endogenous pain control Pharmacol Exp Ther. 1953 Mar; 107: 385-393.
systems: brainstem spinal pathways and endor- 24) WANG L, MILNE B, JHAMANDAS K. Involvement of ex-
phin circuitry. Ann Rev Neurosci 1984; 7: 309- citatory amino acid pathway in the expression of
338. precipitated opioid withdrawal in the rostral ven-
12) YANG D, YANG S, ZHANG Y, LIU Y, MENG X, LIANG Z. trolateral medulla: an in vivo voltametric study.
Metabolic profiles of three related Salvia species. Brain Res 1995; 697: 130-142.
Fitoterapia 2009; 80: 274-278. 25) HOSSEINZADEH H, STONE TW. Adenosine in the cen-
13) WINSTON D. The use of botanicals in eclectic pedi- tral nervous system. Med J Isl R Iran 1996; 9:
atrics. J Am Herbalists Guide 2004; 3: 59-64. 361-368.
14) R ECHINGER KH, 1982. S ALVIA . In: Flora Iranica, 26) LEE CM, WONG HN, CHUI KY COANG TF, HON PM,
Rechinger KH and IC Hedge Akademische Druck CHANG HM. (1991). Miltrione, a central benzodi-
and Verlagsanstalt, Graz, Austria, p. 439. azepine receptor partial agonist from a chinese
15) IMANSHAHIDI M, HOSSEINZADEH H. The pharmacologi- medicinal herbs Salvia militorrhiza. Neurosci Lett
cal effects of Salvia species on the central ner- 1991; 127: 241-273.
vous system. Phytother Res 2006; 20: 427-437. 27) RUTHERFORD DM, NELSON MP, HANSEN SK, WITT MR,
16) BARICEVIC D, SOSA S, DELLA LR, TUBARO A, SIMONOVS- BERGENDROFF O, STERNER O. Isolation and identifica-
KA B, KRASNA A, ZUPANCIC A. Topical anti-inflamma- tion from Salvia officinalis of two diterpenes which
tory activity of Salvia officinalis L. leaves: the rel- inhibit t-butylbicyclophosphoro [35S] thionate
evance of ursolic acid. J Ethnopharmacol 2001; binding to chloride channel of rat cerebrocortical
75: 125-132. membranes in vitro. Neurosci Lett 1992; 135:
224-226.
17) HERNANDEZ-PEREZ M, RABANAL RM, DE LA TORRE MC,
R ODRIGUEZ B. Analgesic, antiinflammatory, an- 28) ZARRINDAST MR, MOHAJERI S. Influence of ATPde-
tipyretic and haematological effect of pendent K+ channels on nicotine-induced inhibi-
aethiopinone, ano-naphthoquinone diterpenoid tion of withdrawal in morphine-dependent mice.
from Salvia aethiopis roots and two hemisynthet- Eur J Pharmacol 2006; 552: 90-98.
ic derivatives. Planta Med 1995; 61: 505-509. 29) TAO Y. Effect of Salvia miltiorrhiza compositae on
18) HOSSEINZADEH H, YAVARI M. Anti-inflammatory ef- phosphoinositides metabolism in acute myocar-
fects of Salvia leriifolia Benth. leaf extract in dial ischemia. Chang Kvo Chang His Chieh Ho
mice and rats. Pharm Pharmacol Let 1999; 9: 60- Iso Chin 1993; 13: 354-355.
61. 30) F UNDYTUS ME, C ODERRE TJ. Effect of activity at
19) HOSSEINZADEH H, LARI P. Effect of Salvia leriifolia metabotropic, as well as ionotropic (NMDA), glu-
extract on morphine dependence in mice. Phy- tamate receptors on morphine dependence. Br J
tother Res 2000; 14: 384-387. Pharmacol 1994; 113: 1215-1220.
20) HOSSEINZADEH H, HADDADKHODAPARAST MH, SHOKO- 31) THOMAS JM, FRAZIER JS, HU ZW, HOFFMAN BB. Phos-
HIZADEH H. Antihyperglycemic effect of Salvia leri- phorylation of cyclic AMP response element bind-
ifolia Benth. leaf and seed extract in mice. Irn J ing protein and induction of c-fosgene expression
Med Sci 1998; 23: 74-80. on withdrawal from chronic treatment with carba-
21) MARSHALL L, GRAHAME-SMITH DG. Evidence against chol in NG108-15 cells. Mol Pharmacol 1995; 48:
a role of brain 5-HT in the development of physi- 593-600.
cal dependence upon morphine in mice. J Phar- 32) L OOMIS TA. Essential of Toxicology. Lea and
macol Exp Ther 1994; 197: 63-64. Febiger: Philladelphia, 1968; pp. 67-78.
22) HARBORNE JB. Phytochemical methods: A guide to 33) ZARGARI A. Medicinal Plants, Vol. 4. Tehran Univer-
modern technique of plant analysis. Chapman sity Press: Tehran, 1990; pp. 1-57.
42You can also read
