@2025 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2015;21(2):121-128

Background
… [1-3] The consumption of non-steroidal anti-inflammatory medications is the most common method for the temporary relief of symptoms of inflammation, but their continued use for the treatment of chronic inflammation is associated with severe side effects [4]. … [5, 6] The components of the essence and extract of Salvia Multicaulis contain borneol, bornil asetat camphor and camphene. The most compounds in the extract are polyphenols and flavonoids. The essence and extracts of this plant has antioxidant activity, but the effect of the plants’ extract on free radicals removal is more essential than the essence [7]. … [8] Antibacterial activity of the plant essence against ‘translucens pv. Cerealis xanthomonas bacteria’ can be attributed to camphor, cineole, thujene and borneol derivatives [9]. Essence of Salvia multicaulis and especially some of its components such as cineole, thujene and camphor have antimicrobial and anti-cancer properties [10, 11]. … [12-16]
Previous Studies
The extract of Salvia multicaulis leaves has analgesic properties in both primary and secondary pain stages, so that 50, 100 and 200 mg/kg of body weight leaf extracts significantly reduce the appeared responses in the initial phase of induced pain by the formalin test. In addition, like morphine, the extract could significantly reduce the initial phase of induced pain by formalin test. In the second phase of pain, also, different concentrations of extract, dependent on the dose, have reduced the pain induced by the formalin test [8].
Aim(s)
The aim of this study was to evaluate the analgesic and anti-inflammatory effects of hydro-alcoholic extract of Salvia Multicaulis.
Research type
This is an experimental study.
Research society, place and time
Approximately 2-month rats weighed 200 to 250gr were studied in the Research Laboratory of Animal Physiology, Ferdowsi University of Mashhad (Iran).
Sampling method and number
42 animals were studied.
Used devices&materials
All animals were kept at the same environment with 12-hour light/darkness cycle at 22 ± 2°C and they had no restricted access to food and water. Salvia Multicaulis was gathered from the Height Mountains of Kermanshah and Islamabad Nowakuh. Extraction was performed using extraction method with ethanol 70%. 42 rats were randomly divided into six 7-rat groups including control group (received no substance), sham (ethanol solvent combination intra-peritoneal injection, Tween 80 and saline at 1: 1: 8 ratio) and four groups receiving 50, 100, 200 and 400 mg/kg Salvia Multicaulis extract via intra-peritoneal injection. Formalin test was used to measure chemical pain [17-19]. To this end, 0.05ml formalin 2.5% were injected subcutaneously into the hind foot and animal behavior has been studied for an hour. The degree of pain was recorded every 15 seconds. Then, average degree of pain was measured every 5 minutes. Zero was for the time when there was no pain and the animal’s foot was on the ground; one score was for the time when only foot toe was on the ground; two scores were assigned when the animal hold his foot totally above the ground and three scores were assigned when animal licked or bit the foot [17].To measure thermal pain, the test of withdrawal of tail was used [20]. In this test, the light intensity of the device on the middle third of the animal's tail was set in such a way that the mean time for withdrawal of tail was 4 to 6 seconds. This test was performed three times with the interval of one minute in the middle third of the tail and the mean data was considered as the threshold for incidence of pain (delay in the withdrawal of tail) for each animal. To compare the groups, the results were reported as the maximum possible effect percent (MPE%). To measure the degree of inflammation, inflammation-induced paw edema volume measurement was applied. In this method, animal’s feet below the ankle, which was marked by a marker, were immersed into the mercury column that was placed on a digital scale and the number showed by the scale was recorded. Then after 30 minutes of intra-peritoneal administration, to induce inflammation, 0.5cc formalin 2.5% was injected subcutaneously into the animal’s foot. The secondary foot volume, also, was measured using plethysmometery method and the edema volume induced by formalin was calculated using the formula “(primary foot volume -secondary foot volume) divided by 13.6 (density of mercury)”. To analyze data, SPSS 16 software was used. Analysis of Variance with Repeated Measure was used in analysis of variance. Student T-test (to compare the mean scores) and Probity Assessment (to calculate LD 50) were used. … [21]
Findings by Text
The amount of LD50, 72 hours after intra-peritoneal administration, was 7356 mg/kg of body weight. Degree of chemical pain showed no significant difference between sham and control group. Intra-peritoneal administration of 50, 100, 200 and 400 mg/kg of hydoralchohalic extract of purple Salvia Multicaulis reduced the chemical pain induced by formalin in foot in both the first (neurogenic) and the second (inflammatory) formalin test compared to sham group (Diagram 1). Degree of thermal pain was no significantly different in control and sham groups. Intra-peritoneal administration of 200 and 400 mg/kg of purple Salvia Multicaulis extract induced hyperalgesia compared to control group (Diagram 2). The results of measuring the foot volume changes induced by formalin in the plantar in sham and control groups showed no effect of solvents on foot edema and lack of significant differences between both groups. Comparison between the effects of different doses of hydro-alcoholic extract of the plant and control group showed that all doses of intra-peritoneal administration led to a significant decrease in the volume of foot inflammatory edema. In addition, the amount of inflammation in experimental groups had an inverse relation with dose increase compared to control group (Diagram 3). Comparison between the differences of paw volume due to plantar formalin injection in 50 and 100mg/kg groups showed no significant difference between the groups, while a significant difference was observed between other doses. The results of thermal pain threshold measurement via tail withdrawal test showed a significant difference between 400mg/kg group and 50mg/kg group. No significant difference was observed between the groups.
Main comparison to the similar studies
Intra-peritoneal administration of Salvia Multicaulis extract had analgesic effect during formalin test in both first and second phases of the test. Antioxidant effects of the extract and essence of this plant are higher than synthetic antioxidants like BHT (Butilhidroxitolueno), ascorbic acid and curcumin [22]. Analgesic activity of the abundance of alkaloids, terpenoids, flavonoids and lactones has been shown [8]. … [23-34]
Suggestions
To determine the exact mechanism of this extract, VRL-1 antagonists and agonists receptors must be used.
Limitations
There was no important limitation.
Conclusions
Intra-peritoneal administration of Salvia Multicaulis extract has anti-nociceptive effect, leading to chemical pain reduction in formalin test.
Acknowledgments
Research Department of Ferdowsi University of Mashhad is appreciated.
Conflict of interest
Non-declared
Ethical Permissions
All experiments were performed in accordance with the ethical instructions about laboratory animals.
Funding Sources
This study was funded by Ferdowsi University of Mashhad.

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