1 The inflammatory reaction is a part of the defense mechanisms of natural immunity also known as innate immunity which is based on the establishment of an early inflammatory reaction to aggressions (e.g., infection or tissue injury). The symptoms of inflammation are redness, heat, and swelling accompanied by pain [1,2]. Fever is the increase in body temperature above the normal physiological threshold that can be caused by infection or inflammation [3]. The inflammatory process is associated with increased production of prostaglandins [4].
2 Nonsteroidal anti-inflammatory drugs (NSAIDs) are often prescribed as treatment of pain, fever, inflammation, and rheumatic disorders. However, their long-term therapeutic use is often associated with side effects such as gastrointestinal ulcers and renal failure [5].
3 The majority of the world's population uses plants in traditional medicine; indeed, plants are capable to produce several classes of chemical constituents with interesting biological activities [6,7]. In this context, the use of natural resources, particularly medicinal plants, is an alternative way to discover effective drugs with fewer side effects [8].
4 Inulaviscosa(L.) Aiton = Dittrichiaviscosa(L.) Greuter from the Asteraceae family is in abandoned and plowed fields in the Mediterranean region [9,10]. This species is used in folk medicine as an antiphlogistic [11], antipyretic [12], antimicrobial [13], and antifungal [10].
5 Various compounds have been isolated and identified fromInulaviscosa(L.) such as flavonoids (7-O-Methylaromadendrin), monoterpenes, triterpenoids, and sesquiterpenes (Tomotonsinn, costic acid). Further, these compounds have been reported to have anti-inflammatory, antidiabetic, antioxidant, and antitumor properties [14].
6 In this work, we were interested in studying the anti-inflammatory, analgesic, and antipyretic activities of some extracts of Inulaviscosa(L.) from Algeria. We chose decoctions for their frequent use in traditional medicine and methanolic extracts.
7 All chemicals and reagents used in this study were of analytical grade. Diclofenac, ibuprofen, carrageenan, and dried brewer's yeast (Saccharomycescerevisiae) were provided by SAIDAL (National Pharmaceutical industry, Algeria), and chemical solvents and reagents (acetic acid, n-hexane, methanol (100%), Folin–Ciocalteu, aluminum chloride), and phytochemical markers (Gallic acid and Quercetin) were purchased from Sigma-Aldrich USA.
8 Instruments: Spectrophotometer (Lambda 25 PerkinElmer, USA), rotary evaporator (300b Stuart, UK), and lyophilizer (Edwards Alto Vuoto, Italy).
9 Leaves and flowers (capitula) of Inulaviscosa(L.) were collected, respectively, in April and November 2014 from Tipaza (northern Algeria). Identification of the plant was confirmed by the National Superior Agronomic School (ElHarrach, Algeria). The two parts were air-dried separately at room temperature under shade and were later ground to fine powder and stored until use.
10 Male and female Swiss albino mice (25–30g) were used to study the acute toxicity, anti-inflammatory, and analgesic activities; females were nulliparous and not pregnant. Male Wistar rats (150–200g) were used to study the antipyretic activity. The animals were provided by the laboratory of pharmaco-toxicology of Antibiotical Saidal Company (Medea, Algeria) and the Center of Research and Development of Saidal Company (El Harrach, Algeria). Mice and rats were housed under standard conditions of temperature (22 ± 2°C), relative humidity (50 ± 15%), and photoperiod (12h light and dark cycle). Commercial pellet diet and water were provided ad libitum. They were acclimatized to laboratory conditions for at least one week before testing. The animals were fasted overnight prior to each experiment but had free access to water.
11 The powdered samples of leaves and flowers (50g) were successively extracted with n-hexane (delipidation to eliminate fatty acids) and methanol (100%) using a Soxhlet apparatus. The different solvent extracts were filtered through a Whatman filter paper no4and concentrated under vacuum using a rotary evaporator to obtain n-hexane and methanolic dry extracts of leaves and flowers [15].
12 For the preparation of 10% decoctions, 50g of powdered samples of leaves and flowers were boiled with 500ml of distilled water for 15min. The mixtures were filtered through a Whatman filter paper no4. Decoctions were then concentrated under reduced pressure and lyophilized [16].
13 Total phenolic content was determined with the Folin–Ciocalteu reagent as described by Singleton and Rossi [17]. 200μl of each extract (1mg/ml) was mixed with 1ml of Folin–Ciocalteu reagent (diluted 10times) for 4min, and 800μl of saturated sodium carbonate solution (about 75g/l) was added into the reaction mixture. The absorbance readings were taken at 760nm after incubation at room temperature for 30min. Gallic acid was used as a reference standard, and the results were expressed as milligram gallic acid equivalent per g of dry extract (mg GA. Eq/g DE). Samples were prepared in triplicate for each analysis.
14 The flavonoid content in the plant extracts was determined using the aluminum chloride (AlCl3) method [18]. 1ml of a 2% solution of AlCl3 was added to 1ml of each extract (1mg/ml). The mixture was incubated for 10min at room temperature, and the absorbance was read at 430nm. The same procedure was repeated for the standard solution of Quercetin to generate a calibration curve. The flavonoid content for each extract was expressed in milligram Quercetin equivalent per gram of dry extract (mgQ. Eq/gDE). Samples were prepared in triplicate for each analysis.
15 The acute toxicity study was performed on Swiss albino mice of either sex selected randomly. Males and females were kept in separate cages until the end of the experiment. A single dose (2,000mg/kg) of methanolic extracts and decoctions of leaves and flowers of Inulaviscosa(L.) were administered orally (p.o.) to different groups containing six mice each (three males and three females). The control group received saline. Mice were kept under regular observation for 48h for any adverse effects, including mortality. Other behavioral changes and parameters, food intake, water intake, diarrhea, and locomotor activity were also monitored. Observations were further extended up to 14days for any signs of mortality [19].
16 The anti-inflammatory study was carried out using the carrageenan-induced paw edema in mice method as described by Winter etal. [20]. Mice were randomized into 14groups of six mice each
- Groups 1, 2, and 3: received Leaf Methanolic Extract (LME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 4, 5, and 6: received Leaf Decoction (LD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 7, 8, and 9: received Flower Methanolic Extract (FME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 10, 11, and 12: received Flower Decoction (FD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Group 13: received diclofenac as a reference drug at the dose of 50mg/kg (p.o.); and
- Group 14: received saline solution as a control (p.o.).
17 After 30min, inflammation was induced by injecting 0.1ml of an edematogenic agent: carrageenan 1% (w/v) suspension in isotonic saline, into the sub-planter surface of the left hind paw of the mouse. Paw thickness was measured before application of the inflammatory substance and at 0, 1, 2, 3, and 4h after induction of inflammation. The difference in footpad thickness was measured by a Vernier caliper.
18 The percentage of inhibition of inflammatory reaction (I%) was determined for each animal by comparison to the control and calculated by the formula:
19 The acetic acid-induced writhing method described by Koster etal. [21] was adopted for evaluation of analgesic activity. Writhing movement was recognized as contraction of abdominal muscle together with the stretching of hind limbs. Mice were divided into 14groups of six animals each:
- Groups 1, 2, and 3: received Leaf Methanolic Extract (LME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 4, 5, and 6: received Leaf Decoction (LD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 7, 8, and 9: received Flower Methanolic Extract (FME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 10, 11, and 12: received Flower Decoction (FD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Group 13: received diclofenac as a reference drug at the dose of 50mg/kg (p.o.); and
- Group 14: received saline solution as a control (p.o.).
20 After 30min, writhing was induced in mice by intraperitoneal injection of 0.2ml of 1% acetic acid solution. 5min after acetic acid injection, the animals were observed and the number of writhes for each mouse was counted in 15min. The response of treated groups was compared with animals in the control group.
21 The percentage inhibition of writhes (I%) was calculated by the following formula:
22 The antipyretic activity was evaluated using Brewer's yeast-induced pyrexia in rats according to the method of Asongalem etal. [22]. Before yeast injection, basal rectal temperature of rats was recorded using a digital clinical thermometer; thereafter, pyrexia was induced by subcutaneous injection of 10ml/kg of the 15% yeast solution into the rat's dorsum region. The rectal temperature of each rat was measured 18h after injection. Only rats that showed an increase in temperature of 0.5–1°C were used for experiments. Rats were divided into 14groups of six animals each:
- Groups 1, 2, and 3: received Leaf Methanolic Extract (LME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 4, 5, and 6: received Leaf Decoction (LD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 7, 8, and 9: received Flower Methanolic Extract (FME) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Groups 10, 11, and 12: received Flower Decoction (FD) at the doses of 400, 600, and 800mg/kg, respectively (p.o.);
- Group 13: received ibuprofen as a reference drug at the dose of 100mg/kg (p.o.); and
- Group 14: received saline solution a control (p.o.).
23 The rectal temperature (T °C) of all rats was taken at 1, 2, 3, and 4h after drug and extracts administration and was compared with the control group.
24 The percentage inhibition of pyrexia (I%) was calculated by the following formula:
25 The experimental results are expressed as mean ± standard error of the mean (S.E.M.). The statistical significance of the differences between groups was determined by one-way analysis of variance (ANOVA) followed by Dunnett's multiple range test for In vivo studies and Tukey's multiple range test for In vivo and phytochemical studies. P < 0.05 was considered statistically significant, using XLSTAT 2014 software (Pro statistical software, Addinsoft, Paris, France).
26 The highest extraction yield was found in LME (27.26%). The total phenolic content using the Folin–Ciocalteu's method (y = 0.004x + 0.048;R2 = 0.999) (Table 1) calculated in LME (387.33mg GAE/g DE) was higher than those found in LD (310.33mg GA. Eq/g DE), FME (251.33mg GA. Eq/g DE) and FD (208.33mg GA. Eq/g DE) with P < 0.05.
Table 1
| Extracts | Yield% | Total phenolicmg GA. Eq/g DE | Total flavonoidmg Q. Eq/g DE |
|---|---|---|---|
| LME | 27.26 | 387.33 ± 0.88a | 141 ± 1.53a |
| FME | 16.47 | 251.33 ± 0.67c | 128.33 ± 0.67b |
| LD | 18.5 | 310.33 ± 1.67b | 115 ± 1c |
| FD | 13.15 | 208.33 ± 0.88d | 109.67 ± 0.33d |
mg GA. Eq/g DE: milligram gallic acid equivalent per g ofdry extract; mg Q. Eq/g DE: (milligram Quercetin equivalent per gram ofdry extract). Values forLME (Leaf Methanolic Extract), LD (Leaf Decoction), FME (Flower Methanolic Extract), andFD (Flower Decoction) are reported asmean ± SEM forN = 3. Different superscript letters within thesame column indicate significant statistical difference (P < 0.05) according toaone-way ANOVA followed byTukey's multiple test comparisons
27 The total flavonoid content (y = 0.004x–0.019;R2 = 0.999) (Table 1) of LME, LD, FME, and FD was 141mg Q. Eq/g DE; 115mg Q. Eq/g DE; 128.33mg Q. Eq/g DE; and 109.67mg Q. Eq/g DE ± 0.33, respectively. The highest content of flavonoid was also observed in LME with P < 0.05.
28 Bioactive secondary metabolites are essentially found in plants. These bioactive molecules have a great influence on the healthcare system [23].
29 Total phenolic and flavonoid content of methanolic extracts found in our study was higher than those found by Brahmi etal. [24] and Abu-Qatouseh etal. [25] in Algeria; Mahmoudi etal. [26] and Rhimi etal. [27] in Tunisia; Chahmi etal. [28] in Morocco; Salim etal. [29] in Palestine; and Orhan etal. [30] in Turkey. They observed less amounts of phenols and flavonoids in LME and FME as compared to our results. These results can suggest that Inulaviscosa(L.) leaves and flowers harvested in Tipaza are rich in phenolic compounds. According to Ksouri etal. [31] and Medeni etal. [32], total phenolic content of plants depends on the studying species, the organ used, the environment, and the extraction solvents.
30 The oral administration of LME, LD, FMD, and FD at the dose of 2,000mg/kg did not produce any mortality or signs of acute toxicity during the 14days of observation.
31 The anti-inflammatory effect of methanolic extracts and decoctions of Inulaviscosa(L.) leaves and flowers evaluated by the carrageenan-induced paw edema is shown in table2, table3, and figure1. After 4hours, oral administration of LME at the different doses of 400, 600, and 800mg/kg reduced significantly mice paw edema as compared to the control group with a percentage of edema reduction of 49.06%,73.58%, and 90.57%, respectively. Concerning the activity of LD at the doses of 400, 600, and 800mg/kg, we observed reduction of the paw edema by 32.08%,43.40%, and 69.81%, respectively. As shown in table3and figure1, FME at the doses of 400, 600, and 800mg/kg inhibited significantly paw edema (P < 0.001) with 41.51%,60.38%, and 77.36% of inhibition, respectively. FD at 400, 600, and 800mg/kg also reduced mice paw edema with 28.30%,35.85%, and 58.49% of inhibition, respectively. All the tested extracts showed significant reduction of mice paw edema comparing to the control group not in a dose-related manner. Furthermore, paw edema inhibition obtained with the oral administration of LME at 600and 800mg/kg, LD at 800mg/kg, and FME at 800mg/kg was not significantly different from that of diclofenac (79.25%).
Table 2
| Extracts | Change inpaw thickness (mm) | ||||
|---|---|---|---|---|---|
| 0h | 1h | 2h | 3h | 4h (% ofinhibition) | |
| Control | 0.34 ± 0.04 | 0.36 ± 0.03 | 0.4 ± 0.03 | 0.44 ± 0.04 | 0.53 ± 0.05 |
| Diclofenac (50mg/kg) | 0.31 ± 0.04 | 0.27 ± 0.02* | 0.22 ± 0.03*** | 0.16 ± 0.03*** | 0.11 ± 0.03*** (79.25) |
| LME (400mg/kg) | 0.29 ± 0.009 | 0.31 ± 0.01 | 0.3 ± 0.01** | 0.25 ± 0.009*** | 0.27 ± 0.01*** (49.06) |
| LME (600mg/kg) | 0.27 ± 0.01 | 0.23 ± 0.008*** | 0.18 ± 0.008*** | 0.17 ± 0.09*** | 0.14 ± 0.009*** (73.58) |
| LME (800mg/kg) | 0.28 ± 0.02 | 0.2 ± 0.02*** | 0.12 ± 0.009*** | 0.09 ± 0.006*** | 0.05 ± 0.02*** (90.57) |
| LD (400mg/kg) | 0.3 ± 0.01 | 0.35 ± 0.01 | 0.32 ± 0.02* | 0.32 ± 0.01** | 0.36 ± 0.01*** (32.08) |
| LD (600mg/kg) | 0.32 ± 0.04 | 0.34 ± 0.005 | 0.31 ± 0.02* | 0.28 ± 0.02*** | 0.3 ± 0.02*** (43.40) |
| LD (800mg/kg) | 0.32 ± 0.04 | 0.28 ± 0.03* | 0.21 ± 0.02*** | 0.19 ± 0.02*** | 0.16 ± 0.01*** (69.81) |
Values are reported asmean ± SEM forN = 6per group fordiclofenac, LME (Leaf Methanolic Extract), andLD (Leaf Decoction). Statistical significance was determined using aone-way ANOVA followed byDunnett's test (*P < 0.05; **P < 0.01; ***P < 0.001) comparing theexperimental group tothecontrol group
Table 3
| Extracts | Change inpaw thickness (mm) | ||||
|---|---|---|---|---|---|
| 0h | 1h | 2h | 3h | 4h (% ofinhibition) | |
| Control | 0.34 ± 0.04 | 0.36 ± 0.03 | 0.4 ± 0.03 | 0.44 ± 0.04 | 0.53 ± 0.05 |
| Diclofenac (50mg/kg) | 0.31 ± 0.004 | 0.27 ± 0.02* | 0.22 ± 0.03*** | 0.16 ± 0.03*** | 0.11 ± 0.03*** (79.25) |
| FME (400mg/kg) | 0.27 ± 0.02 | 0.34 ± 0.03 | 0.37 ± 0.05 | 0.29 ± 0.02** | 0.31 ± 0.02*** (41.51) |
| FME (600mg/kg) | 0.33 ± 0.05 | 0.27 ± 0.02* | 0.25 ± 0.03** | 0.24 ± 0.03*** | 0.21 ± 0.02*** (60.38) |
| FME (800mg/kg) | 0.29 ± 0.005 | 0.24 ± 0.03** | 0.19 ± 0.02*** | 0.16 ± 0.02*** | 0.12 ± 0.009*** (77.36) |
| FD (400mg/kg) | 0.30 ± 0.01 | 0.36 ± 0.02 | 0.4 ± 0.05 | 0.39 ± 0.04 | 0.38 ± 0.02** (28.30) |
| FD (600mg/kg) | 0.32 ± 0.04 | 0.36 ± 0.02 | 0.35 ± 0.04 | 0.32 ± 0.01* | 0.34 ± 0.03*** (35.85) |
| FD (800mg/kg) | 0.28 ± 0.01 | 0.33 ± 0.04 | 0.28 ± 0.01* | 0.25 ± 0.04*** | 0.22 ± 0.03*** (58.49) |
Values are reported as mean ± SEM for N = 6 per group for diclofenac, FME (Flower Methanolic Extract), and FD (Flower Decoction). Statistical significance was determined using a one-way ANOVA followed by Dunnett's test (*P < 0.05; **P < 0.01; ***P < 0.001) comparing the experimental group to the control group
Fig. 1
Values are reported asmean ± SEM with N = 6 per group. Different superscript letters inthesame figure indicate significant statistical difference (P < 0.05) according toaone-way ANOVA followed by Tukey's multiple test comparisons
32 Carrageenan-induced paw edema is widely used as a working model of inflammation in the search for new anti-inflammatory drugs [33]. The edema formation is a biphasic event, the release of histamine, serotonin, and bradykinin occurs in the first phase, and the second phase is associated with the production of prostaglandin, protease, and lysosomal enzymes [34–36]. The carrageenan-induced paw edema test is effectively controlled with the arachidonate cyclooxygenase (COX) inhibitors due to its COX-dependent mechanism [37]. Thus, from the observed results, it is suggested that Inulaviscosa(L.) extracts may process arachidonate COX inhibitory property.
33 Studies carried out by Mañez etal. [38,39] and Hernandez etal. [40,41] showed that sesquiterpenes (illicic acid, hydroxycostic acid, inuviscolide) and phenolic compounds such as flavonoids (sakurantin, 7-O-Methylaromadendrin) present in Inulaviscosa inhibited the swelling induced by TPA (12-O-tetradecanoylphorbol-13-acetate) in models of skin inflammation. Furthermore, inuviscolide a sesquiterpene lactone from Inulaviscosa caused a significant reduction of leukocyte infiltration and inhibited cyclooxygenase 1 [39]. The possible presence of sesquiterpenes in the tested extracts could partly explain their anti-inflammatory effect. Furthermore, the inhibition of carrageenan-induced paw edema by Inulaviscosa(L.) extracts could be due to the inhibition of the cyclooxygenase enzyme and subsequent inhibition of prostaglandin synthesis.
34 The oral administration of LME, LD, FME, and FD exhibited significant inhibition of the writhes not in a dose-dependent manner. As observed in table4and figure2, pretreatment with LME at doses of 400, 600, and 800mg/kg reduced the number of writhes caused by the injection of acetic acid, with a percentage of analgesia of 85.67%,89.80%, and 93.39% at the doses of 400, 600, and 800mg/kg, respectively. LD at the same doses also reduced the abdominal writhing response by 70.79%,78.51%, and 85.4%, respectively. Concerning the FME and FD, both showed a significant decrease of writhes compared to the control group with percentage of analgesia of 89.53% and 79.34%, respectively, for FME and FD at the dose of 800mg/kg. The effect observed with LME at 400, 600, and 800mg/kg, LD at 800mg/kg, FME at 400, 600, and 800mg/kg, and FD at 800mg/kg was comparable to that of the reference drug (Diclofenac at 50mg/kg; 88.17%).
Table 4
| Extracts | Number ofwrithes | % ofanalgesia |
|---|---|---|
| Control | 60.5 ± 3.05 | |
| Diclofenac (50mg/kg) | 7.17 ± 1.28*** | 88.15 |
| LME (400mg/kg) | 8.67 ± 1.17*** | 85.67 |
| LME (600mg/kg) | 6.17 ± 1.14*** | 89.80 |
| LME (800mg/kg) | 4 ± 0.78*** | 93.39 |
| LD (400mg/kg) | 17.67 ± 1.54*** | 70.79 |
| LD (600mg/kg) | 13 ± 1.03*** | 78.51 |
| LD (800mg/kg) | 8.83 ± 0.96*** | 85.4 |
| FME (400mg/kg) | 11.17 ± 1.3*** | 81.54 |
| FME (600mg/kg) | 8 ± 0.84*** | 86.78 |
| FME (800mg/kg) | 6.33 ± 1.12*** | 89.53 |
| FD (400mg/kg) | 21.67 ± 1.59*** | 64.19 |
| FD (600mg/kg) | 18.17 ± 1.28*** | 69.97 |
| FD (800mg/kg) | 12.5 ± 1.07*** | 79.34 |
Values are reported asmean ± SEM forN = 6per group forLME (Leaf Methanolic Extract), LD (Leaf Decoction), FME (Flower Methanolic Extract), andFD (Flower Decoction). Statistical significance was determined using aone-way ANOVA followed byDunnett's test (***P < 0.001) comparing theexperimental group tothecontrol group
Fig. 2
Values are reported asmean ± SEM with N = 6 per group. Different superscript letters inthesamefigure indicate significant statistical difference (P < 0.05) according toaone-way ANOVA followed by Tukey's multiple test comparisons
35 Acetic acid writhing test was used in order to evaluate the peripheral analgesic action of Inulaviscosa(L.) This method has been used by different research groups to determine the antinociceptive effect of natural compounds [42,43]. Abdominal writhes are manifested by the curving of the back, and the extension of hind limbs [44]. This test induced pain by liberation of various endogenous substances which excite the peripheral nociceptors [45]. Some mediators (histamine, bradykinin, serotonin, or prostaglandins) and cytokines produced in the peritoneal fluid cause an elevation of the vascular permeability and induce the stimulation of the nervous terminal of nociceptive fibers [43,46]. The reduction in the number of writhes induced by Inulaviscosa(L.) methanolic extracts and decoctions suggests that the antinociceptive effect may be peripherally mediated by the inhibition of synthesis and/or liberation (release) of prostaglandins.
36 The results of the antipyretic activity of Inulaviscosa(L.) extracts on rats are illustrated in table5, table6, and figure3. The subcutaneous injection of yeast suspension had elevated the rectal temperature after 18h. Animals in the control group had a significant increase in rectal temperature throughout the experimental period, while those treated with ibuprofen (100mg/kg) showed significant decrease of rectal temperature during the experimental period with maximum attenuation at 4h. The antipyretic activity of LME of Inulaviscosa showed a significant reduction of pyrexia atthedoses of 600and 800mg/kg after the firsthour (P < 0.01; P < 0.001) until the fourthhour, while at the dose of 400mg/kg LME showed significant reduction of rectal temperature after 2h. Maximum diminution of hyperthermia observed with LME was of 89.51% at 800mg/kg after 4hours which was comparable to the reference drug (91.61%). FME at 600and 800mg/kg produced a significant decrease of rectal temperature after the firsthour of treatment, while FME at 400mg/kg showed a significant decrease in temperature after 3h of treatment. The maximum attenuation of pyrexia observed with the FME was of 87.41% at 800mg/kg after 4h. Inulaviscosa LD and FD demonstrated significant antipyretic activity at test dose of 800mg/kg throughout the assessment times (1–4h) with a percentage of inhibition of pyrexia of 77.62% and 69.23%, respectively. The effect of the extracts was not in a dose-dependent manner. The obtained results with the oral administration of LME at 600and 800mg/kg, FME, and LD at 800mg/kg was not significantly different than to those of ibuprofen at 100mg/kg (91.61%).
Table 5
| Extracts | Temperature (°C) | |||||
|---|---|---|---|---|---|---|
| Normal | 18h after yeast | 1h | 2h | 3h | 4h (% reduction) | |
| Control | 37.17 ± 0.18 | 38.35 ± 0.27 | 38.48 ± 0.25 | 38.43 ± 0.24 | 38.57 ± 0.23 | 38.6 ± 0.18 |
| Ibuprofen (100mg/kg) | 37.15 ± 0.13 | 38.18 ± 0.15 | 37.67 ± 0.14 ** | 37.25 ± 0.16 *** | 37.17 ± 0.17 *** | 37.27 ± 0.17*** (91.61) |
| LME (400mg/kg) | 37.12 ± 0.13 | 38.42 ± 0.19 | 38.07 ± 0.16 | 37.65 ± 0.11 ** | 37.42 ± 0.12*** | 37.57 ± 0.17 *** (68.53) |
| LME (600mg/kg) | 37.12 ± 0.15 | 38.32 ± 0.17 | 37.65 ± 0.13 ** | 37.48 ± 0.16*** | 37.42 ± 0.18*** | 37.4 ± 0.14*** (80.42) |
| LME (800mg/kg) | 37.1 ± 0.16 | 38.38 ± 0.16 | 37.58 ± 0.16 ** | 37.4 ± 0.18 *** | 37.32 ± 0.19*** | 37.25 ± 0.18***(89.51) |
| LD (400mg/kg) | 37.3 ± 0.16 | 38.48 ± 0.13 | 38.45 ± 0.14 | 38.12 ± 0.14 | 37.98 ± 0.17* | 37.93 ± 0.13*(55.94) |
| LD (600mg/kg) | 37.12 ± 0.14 | 38.25 ± 0.16 | 38.08 ± 0.12 | 37.77 ± 0.17 * | 37.62 ± 0.14** | 37.65 ± 0.15** (62.94) |
| LD (800mg/kg) | 37.2 ± 0.16 | 38.3 ± 0.17 | 37.68 ± 0.13 ** | 37.58 ± 0.12 ** | 37.55 ± 0.12*** | 37.52 ± 0.14*** (77.62) |
Values are reported asmean ± SEM forN = 6per group forLME (Leaf Methanolic Extract) andLD (Leaf Decoction). Statistical significance was determined using aone-way ANOVA followed byDunnett's test (*P < 0.05; **P < 0.01; ***P < 0.001) comparing theexperimental group tothecontrol group
Table 6
| Extracts | Temperature (°C) | |||||
|---|---|---|---|---|---|---|
| Normal | 18h after yeast | 1h | 2h | 3h | 4h (% reduction) | |
| Control | 37.17 ± 0.18 | 38.35 ± 0.27 | 38.48 ± 0.25 | 38.43 ± 0.24 | 38.57 ± 0.23 | 38.6 ± 0.18 |
| Ibuprofen (100mg/kg) | 37.15 ± 0.13 | 38.18 ± 0.15 | 37.67 ± 0.14** | 37.25 ± 0.16*** | 37.17 ± 0.17*** | 37.27 ± 0.17*** (91.61) |
| FME (400mg/kg) | 37.1 ± 0.15 | 38.42 ± 0.13 | 38.4 ± 0.12 | 38.2 ± 0.12 | 37.85 ± 0.16** | 37.7 ± 0.12*** (58.04) |
| FME (600mg/kg) | 37.25 ± 0.13 | 38.37 ± 0.14 | 37.92 ± 0.11* | 37.65 ± 0.12** | 37.5 ± 0.15*** | 37.62 ± 0.15*** (74.13) |
| FME (800mg/kg) | 37.2 ± 0.13 | 38.35 ± 0.11 | 37.77 ± 0.13** | 37.53 ± 0.11*** | 37.42 ± 0.09*** | 37.38 ± 0.11*** (87.41) |
| FD (400mg/kg) | 37.2 ± 0.17 | 38.35 ± 0.17 | 38.47 ± 0.13 | 38.4 ± 0.12 | 38.28 ± 0.13 | 38.1 ± 0.13* (37.06) |
| FD (600mg/kg) | 37.05 ± 0.13 | 38.17 ± 0.13 | 38.31 ± 0.10 | 38.03 ± 0.11 | 37.87 ± 0.11* | 37.73 ± 0.10** (52.44) |
| FD (800mg/kg) | 37.18 ± 0.12 | 38.25 ± 0.15 | 37.82 ± 0.12* | 37.65 ± 0.14** | 37.6 ± 0.15*** | 37.62 ± 0.16*** (69.23) |
Values are reported as mean ± SEM for N = 6 per group for FME (Flower Methanolic Extract) and FD (Flower Decoction). Statistical significance was determined using a one-way ANOVA followed by Dunnett's test (*P < 0.05; **P < 0.01; ***P < 0.001) comparing the experimental group to the control group
Fig. 3
Values are reported asmean ± SEM with N = 6 per group. Different superscript letters inthesame figure indicate significant statistical difference (P < 0.05) according toaone-way ANOVA followed by Tukey's multiple test comparisons
37 Fever is the first sign of disease right from the very beginning of human civilization. The febrile response is regulated by the central nervous system through several mechanisms [47]. There is an increased production of pro-inflammatory mediators such as cytokines like interleukin 1(IL-1) and Tumor Necrosis Factor (TNF) alpha from infected or damaged tissues. These substances elevate the body temperature by increasing the synthesis of prostaglandin E2 (PGE2) near the pre-optic hypothalamus area [48]. Subcutaneous injection of Brewer's yeast causes fever and induces an increase in the synthesis of prostaglandins. It is also considered as a convenient method to study the antipyretic potential of plant extracts [49]. Antipyretic medications such as nonsteroidal anti-inflammatory drugs have the ability to inhibit the prostaglandin synthetase in the hypothalamus [50,51].
38 In this study, Inulaviscosa(L.) leaf and flower methanolic extracts and decoctions caused a significant decrease in body temperature of rats. Therefore, it could be suggested that the antipyretic action of Inulaviscosa(L.) may be related to the inhibition of prostaglandin synthesis in the hypothalamus.
39 Inulaviscosa(L.) extracts contain many compounds suchas phenolic acids, flavonoids, and sesquiterpenes [14,40,41]. It is difficult to predict which of them is responsible for the pharmacological proprieties. Different scientific researchers proved that chemical constituents such as phenolic compounds possess analgesic, anti-inflammatory, and antipyretic activities [52]. The presence of these compounds in Inulaviscosa(L.) can contribute toward their promising anti-inflammatory, analgesic, and antipyretic activities. Several studies have demonstrated the anti-inflammatory and antinociceptive activities of some phenolic acids (chlorogenic acid and dicaffeoylquinic acid derivatives) [53–58], flavonoids (luteolin, quercétine, kaempferol, sakurantin, and 7-O-methylaromadendrin) [40,53,59,60], and sesquiterpenes (illicic acid, hydroxycostic acid, and inuviscolide) [38,39,41]. These compounds or their derivatives have been already identified in Inulaviscosa extracts (L.) [26,27,38–40,61,62], which could explain the anti-inflammatory and analgesic effect obtained in this study. According to Siva etal. [63], plants with analgesic and anti-inflammatory effects have also antipyretic property. Previous studies have related the antipyretic effect to the anti-inflammatory activity of some flavonoids which have an inhibitory action on prostaglandin synthesis [64,65].
40 Furthermore, different species of the genus of Inula have been reported to possess anti-inflammatory (Inulacappa, Inulacuspidata, and Inulabritannica) [66–68], antinociceptive (Inulacuspidata, Inulabritannica, and Inulagraveolens) [67,69,70], and antipyretic effects (Inulagraveolens) [70,71].
41 The obtained results showed that the methanolic extracts ofInulaviscosa leaves and flowers have a good anti-inflammatory, antipyretic, and especially analgesic activity; however, decoctions of leaves and flowers have a moderate anti-inflammatory and antipyretic activity.
42 We found a high content of phenolic compounds in the various extracts, which could explain partly the pharmacological effects of the plant. This study encourages the traditional use and the valorization of Inulaviscosa(L.) as an analgesic, anti-inflammatory, and antipyretic products.
43 In addition, it would be interesting to carry out further research by identifying and testing purified molecules from this plant such as phenolic acids, flavonoids, and sesquiterpenes to understand their mechanism in the treatment of inflammation, pain, and pyrexia.
We wish to thank Lila RABIA, Ph.D. student at the Rensselaer Polytechnic Institute, Department of Chemical and Biological Engineering (Troy, New York) for revising and improving the paper
Conflicts of interests: the authors have no conflicts of interests to declare.
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