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BY 4.0 license Open Access Published by De Gruyter Open Access June 14, 2023

Mineral composition, principal polyphenolic components, and evaluation of the anti-inflammatory, analgesic, and antioxidant properties of Cytisus villosus Pourr leaf extracts

  • Aziz Zouhri EMAIL logo , Naoual El Menyiy , Yahya El-mernissi , Toufik Bouddine , Rafik El-mernissi , Hassan Amhamdi , Abdelhay Elharrak , Ahmad Mohammad Salamatullah , Hiba-Allah Nafidi , Farid Khallouki , Mohammed Bourhia EMAIL logo and Lhoussain Hajji
From the journal Open Chemistry


Cytisus villosus Pourr. (C. villosus) is a medicinal plant belonging to the Fabaceae family, which grows in the Mediterranean area. It is used in traditional medicine against diseases related to inflammation. The objective of the present study was to identify the mineral and polyphenolic composition as well as to evaluate some biological properties including antioxidant, anti-inflammatory, and analgesic activities of C. villosus leaf aqueous extract. The chemical constituents were identified and quantified using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) methods. The antioxidant properties of C. villosus leaves were tested using reducing power (RP), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) assays. The anti-inflammatory potency was evaluated in vitro and in vivo using the albumin denaturation test and the carrageenan test, respectively. Furthermore, the analgesic effect was performed in vivo using tail flick, acetic acid-induced contortion, and plantar tests. Mineralogical analysis revealed that potassium and calcium were the most abundant minerals. The analysis and quantification of the phytochemical composition using UPLC-ESI-MS/MS showed that quinic acid (57.478 ± 1.72 mg/kg) was the major compound of the aqueous extract, followed by salicylic acid (17.38 ± 0.2 mg/kg), isoquercetin (16.895 ± 1.01 mg/kg), and gallic acid (15.914 ± 1.51 mg/kg). The extracts showed potent antioxidant activity for all tests used. The highest antioxidant activity was recorded for the DPPH, ABTS and RP methods, with an IC50 of 3.94 ± 0.09, 2.88 ± 0.07, and 1.94 ± 0.10 μg/mL, respectively. Additionally, using the most frequent analgesic assays, the aqueous extract at a dose of 500 mg/kg exhibited a potent analgesic activity. Notably, an interesting inhibition of albumin denaturation was recorded with an IC50 of 383.94 μg/mL, corroborating the in vivo test. Overall, the results presented here may represent a scientific basis for the traditional use of C. villosus in the treatment of inflammation-related diseases.

1 Introduction

The inflammatory response is a process marked by the stimulation of immune and non-immune cells that defend the body against infections and diseases by eliminating harmful chemicals and promoting tissue repair and regeneration [1]. However, if defensive systems are overwhelmed, inflammation may cause significant alterations in all tissues and organs, which could lead to an increased risk of various chronic pathologies such as cancer, neurodegeneration, diabetes, and hypertension [2].

Inflammation treatments are systemically associated with non-steroidal anti-inflammatory drugs (NSAIDs), which act through the inhibition of cyclooxygenases. However, the use of NSAIDs has been linked with a risk of adverse events [3,4], which have a significant impact on morbidity and represent a substantial increase in healthcare costs [5]. The development of new drugs is consequently an ongoing challenge. Among the options for drug development are plants that constitute a source of bioactive molecules [6].

The plant known as Cytisus villosus Pourr. belongs to the Cytisus genus within the Leguminosae (Fabaceae) family. Typically, this shrub can grow to a height of 1–2 m and features erect stems that give rise to numerous branches. The young branches are angular and covered with long white hairs. The flowers are large, streaked yellow with a papilionaceous corolla. The flowering takes place in April–May [7]. This genus is present in abundance in the Mediterranean basin and used in ancient therapeutic practices as a cure for specific ailments, entailing diabetes and liver diseases, as well as has antioxidant, anxiolytic, diuretic, cardiotonic, and antifungal properties [7,8,9,10,11]. Furthermore, Cytisus villosus Pourr. is rich in phenolic and flavonoid compounds [12,13] Moreover, the plant is linked with numerous biological effects, such as anti-oxidant, anti-inflammatory, and anti-microbial activities [14].

The current study’s objectives are to assess the antioxidant, analgesic, and anti-inflammatory properties of Cytisus villosus Pourr. leaf as well as its chemical and mineral composition.

2 Materials and methods

2.1 Plant material

The Cytisus villosus Pourr. specimen was gathered from the Ketema locality in February 2022 (N: 34°47′56, W: 4°37′44, altitude: 642.1 m) and then identified and cataloged with a reference sample (ANC0039/2022). The plant has been stored at the herbarium of NAMAP (National Agency for Medicinal and Aromatic Plants), Morocco.

2.2 Preparation of plant extracts

The maceration method was used to extract the leaf powder of C. villosus. To obtain the extracts, 100 g of the raw material was macerated at room temperature with either 1 L of distilled water or with ethanol/water (70:30, v/v). The solution was then filtered through Whatman paper no.1 and evaporated using a rotary evaporator at 45°C under reduced pressure. The resulting aqueous and hydroalcoholic extracts had a yield of 19.23% and 15.21% (w/w), respectively, and were stored at 4°C for further use.

2.3 Chemical reagents and standard compounds

Standard compounds, quinic acid, isoquercetin, gallic acid, p-coumaric acid, ferulic acid, salicylic acid, vanillic acid, gentisic acid, 4-OH-phenylacetic acid, caffeic acid, quercetin, 3,4,5-trimethoxycinnamic acid (sinapic acid methyl ether), and avicularin, were purchased from Merck and Carl Roth GmbH (Darmstadt, Germany). Protocatechuic acid, cynaroside, and aromadendrin were purchased from PhytoLab (Dettendorfer, Germany). Astragalin and quercetin were acquired from Extrasynthese (Lyon, France). Gallic acid, Folin–Ciocalteu reagent, catechin, butylated hydroxytoluene (BHT), 2,2′-diphenyl-1-picrylhydrazyl (DPPH), ascorbic acid, and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) were purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Indomethacin, diclofenac sodium, and aspirin were purchased from Pharma5 (Bouskoura, Morocco). The other reagents utilized in the study were of analytical grade and utilized as received.

2.4 Analysis of mineral elements

The mineral composition of C. villosus leaves (silver, aluminum, copper, boron, cobalt, chromium, titanium, iron, calcium, potassium, magnesium, manganese, sodium, nickel, phosphorus, palladium, silicon, tin, zinc, and vanadium) was determined using the calcination method (ICP-AES), as performed previously by Silva et al. [15].

2.5 Determination of the total flavonoid content (TFC), total phenolic content (TPC), and proanthocyanidin content (PC)

The quantification of the TPC was carried out through the Folin–Ciocalteu assay, and the results were reported as milligrams of gallic acid equivalent per gram of dry weight extract (mg GAE/g extract) [16].

The TFC of all extracts was estimated using the protocol detailed by Amezouar et al. [17]. The results were expressed as milligram quercetin equivalent per gram dry weight (mg QE/g extract). The PC content of all extracts was measured following the procedure of Sayah et al. [18]. The results are presented as milligrams of catechin equivalent per gram of dry weight of extract (mg EC/g extract).

2.6 Identification and quantification of phenolic compounds by UPLC-ESI-MS/MS

The acquity liquid chromatography (UPLC) system coupled to a Waters Xevo TQ-S triple quadrupole system (Waters, United States) was used to analyze the aqueous extract of C. villosus according to the methodology described by Zhu et al. [19]. The molecules were identified by checking the typical fragment with those in an in-house developed library of molecules spectra (see supplementary file). The polyphenols were detected in the negative mode of ESI with the deprotonated [M−H]-ion selected as the precursor ion, and optimized mass spectrometry was used to determine the content of the 32 phenolic compounds among the 33 phenolic compounds detected. Table 1 summarizes the precursor-to-product ion transitions.

Table 1

Multiple reaction monitoring transitions for the analyzed phenolic compounds

Compound no. Analyte Retention time (min) Precursor ion (m/z) Product ions (m/z) Ionization mode (+/−)
I Pyrocatechol 5.44 109.2 80.93 ES−
II Gallic acid 4.33 169 125 ES−
169 79
III Protocatechuic acid 6.25 153 109 ES−
IV Vanillic acid 8.92 167 152 ES−
V Gentisic acid 8.06 153 109 ES−
VI Salicylic acid 12.44 137 93 ES−
VII 4-OH-Phenylacetic acid 7.3 151 107 ES−
VIII Quinic acid 2.16 191 85 ES−
191 93
IX p-Coumaric acid 11.5 163 119 ES−
X Caffeic acid 9.58 179 135 ES−
179 107
XI Sinapinic acid 12.2 223 208 ES−
223 164
XII Sinapic acid methyl ether 14.71 237 102 ES−
237 132
XIII Ferulic acid 11.94 193 134 ES−
193 178
XIV Hydroferulic acid 10.12 195 121 ES−
195 93
XV Isoquercetin 13.18 463 300 ES−
463 271
XVI Cynaroside 12.93 447 285 ES−
447 151
XVII Astragalin 14.03 447 284 ES−
447 255
XVIII Quercetin 14.24 447 300 ES−
447 271
XIX Aromadendrin 14.15 287 259 ES−
287 125
XX Avicularin 13.91 433 300 ES−
433 271
XXI Luteolin 16.49 285 133 ES−
285 107
XXII Rutin 12.78 609 300 ES−
609 271
XXIII Kaempferol 17.99 285 93 ES−
285 146
XXIV Naringenin 16.73 271 151 ES−
271 119
XXV Isorhamnetin 17.92 315 300 ES−
315 151
XXVI Taxifolin 12.45 303 285 ES−
303 125
XXVII Quercetin-3-O-glucuronide 13.48 477 301 ES−
477 151
XXVIII Apigetrin 14 431 268 ES−
431 107
XXIX Apigenin 17.74 269 117 ES−
269 149
XXX Phloridzin 13.77 435 273 ES−
435 167
XXXI Quercetin 16.53 301 151 ES−
301 179
XXXII Daidzin 15.49 253 224 ES−
XXXIII Resveratrol 15.39 227 158 ES−

ES−: negative electrospray.

2.7 Antioxidant activity

The ability of our examined extracts to scavenge the DPPH (DPPH) (2,2-diphenyl-1-picrylhydrazyl) radical was measured as described by El Omari et al. [20]. For the preparation of the DPPH solution, 0.005 g of DPPH was dissolved in 200 mL of ethanol (96%) to obtain an absorbance of 0.700 ± 0.01 at 515 nm. Then, 25 μL of the extract at different dilutions (0–1,044 μg/mL) was mixed with 825 μL of the DPPH reagent. The reaction preparation was carefully vortexed and then left in the dark at the laboratory temperature for 30 min. The coloration produced was determined at 517 nm using a spectrophotometer (UV-1700APC, China). The results obtained are compared with the BHT standard. The inhibitory percentage was determined using the following formula:

(1) % I nhibition = ( C ontrol S ample ) C ontrol × 100 .

The ability of C. villosus extracts to inhibit the cation-based ABTS (ABTS•+) radical (2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) was determined by the method described by Miguel et al. [21]. An aqueous solution of ABTS (7 mM) was combined with 2.5 mM potassium persulfate to regenerate ABTS• +. For 16 h, the combination was left at room temperature and in the dark. Using a spectrophotometer, the mixture was adjusted with ethanol (98%) to give an absorbance of 0.70 at 734 nm. Then, 25 μL of the extract at different concentrations was mixed with 825 μL of ABTS•+ reagent. An absorbance measurement was performed at 734 nm after 6 min. The antioxidant standard was ascorbic acid. According to equation (1), the ABTS radical scavenging activity was calculated: The IC50 values were expressed in µg/mL.

The method recommended by Bougandoura and Bendimerad [22] was used to evaluate the reducing potential of the studied extracts, and ascorbic acid was used as a standard reference. The IC50 (µg/mL) of the different assays was determined using different concentrations, in which the % inhibition was between 20 and 80%.

2.7.1 Total antioxidant capacity (TAC)

The phosphomolybdate (PPM) method described in previous study [17] was used to determine TAC. Ascorbic acid was used to establish the calibration range, and the results were expressed as milligrams of ascorbic acid equivalent per gram of crude plant (mg AAE/g E).

2.8 Experimental animal

In this study, adult Wistar rats were used in the experiments. The rats were bred in the NAMAP animal facility (Morocco). All experimental rats were housed in standard environmental conditions (5% humidity, 22 ± 3°C, 55 ±, and 12 h light/dark cycles) and had free access to tap water. Normal (distilled water) and experimental rats (extracts or drugs) were subjected to a fasting period of 16 h before the experiment. All experimental procedures were conducted according to the rules established in the “Guide for the Care and Use of Laboratory Animals,” established by the National Academy of Sciences [23].

2.9 Evaluation of anti-inflammatory activity

To assess the anti-inflammatory potency, the carrageenan-induced rat paw edema test, as per Winter et al. [24], was employed. Each treatment was administered to a group of six rats. The aqueous extract of C. villosus (500 mg/kg bw) was orally administered 30 min before carrageenan injection. Positive and negative controls, indomethacin (10 mg/kg bw) and distilled water (5 mL/kg), were used, respectively. The paw volume was measured before carrageenan injection, and at 1, 3, and 6 h after carrageenan injection using a LE 7500 plethysmometer.

Using equation (2), the anti-inflammatory potency was calculated as a percent inhibition:

(2) Inhibition ( % ) = ( V c V t ) V c × 100 ,

where V c is the average increase in the paw volume of the control group and V t is the average increase in the paw volume of the treatment group.

2.10 Inhibition of albumin denaturation

To study the in vitro anti-inflammatory efficacy of the aqueous extract of C. villosus leaves, we used the albumin denaturation assay according to Lekouaghet et al. [25]. The percentage of protein denaturation was calculated using the following equation:

(3) Inhibition ( % ) = ( Control ( S ample W hite ) ) Control × 100 .

2.11 Analgesic activity study

2.11.1 Writhing test

Contortion provoked by acetic acid was performed as previously reported by Sayah et al. [26]. The weighed rats (180–200 g) were randomized into three groups of six rats each. Group 1 (control) was treated with 0.9% saline, group 2 was pre-treated with aspirin (150 mg/kg bw), group 3 received the C. villosus extract (500 mg/kg bw) 30 min after the treatments, 3.75 mL/kg bw of intraperitoneal acetic acid solution (3%) was injected to cause contortions.

The contortion numbers were counted during a 10 min observation period after the injection of acetic acid. The inhibition (%) of abdominal constrictions was determined as follows :

(4) Inhibition ( % ) = 1 W t W c × 100 ,

where W t and W c represent the contortion numbers in the treated and control group, respectively.

2.11.2 Tail flick test

This test was conducted as previously detailed by Sood et al. [27]. Wistar albino rats (180–200 g) were separated into four groups. Each group consisted of six rats. The first group was considered as a control, the second group was used as a reference (aspirin 150 mg/kg), and the third and fourth groups received aqueous extracts of C. villosus at two doses: 250 and 500 mg/kg p.o.

The tail response was recorded (ANALGESY, METER LE 7106) as the radiant heat sensation from the spine, and a cut-off time of 20 s was maintained. The tail test was performed in each batch before treatment and 30, 60, 90, and 120 min after drug administration.

2.11.3 Plantar test

To determine the nociceptive response to thermal stimuli, we performed a plantar test (UGO BASILE model 37370) following the method previously reported by El Youbi et al. [28] using four groups of six rats each. Group I (normal group): the animals received distilled water. Group II (standard reference group): the animals received aspirin at a dose of 150 mg/kg by gavage. Groups III and IV (experimental group): the animals received C. villosus extract at two different doses (250 and 500 mg/kg p.o.). The latency of the paw withdrawal response was measured automatically at 30, 60, 90, and 120 min.

2.12 Statistical analysis

The findings are reported as mean ± standard error. Graph Pad Prism 8.02 was used to conduct all statistical analyses. Comparisons of C. villosus leaf extracts were performed by analysis of variance (ANOVA) and multiple comparisons were performed by Tukey’s test. Differences were considered significant when P ≤ 0.05.

3 Results and discussion

3.1 Mineral composition

The mineral content of the C. villosus leaf is summarized in Table 2. Importantly, potassium (1130 mg/kg) was the most abundant element, followed by calcium (234 mg/kg), magnesium (90 mg/kg), phosphorus (48.9 mg/kg), and silicon (12.9 mg/kg), while the concentrations of aluminum, sodium, iron, and manganese were 6.07, 2.26, 1.75, and 1.44 mg/kg, respectively. The concentrations of copper, zinc, boron, nickel, chromium, cobalt, silver, tin, vanadium, lead, and titanium were the least abundant. Notably, the potassium, calcium, magnesium, copper, phosphorus, and iron involved in the organism’s defense system against oxidative stress, which may help protect it from ailments [29,30,31].

Table 2

Mineral composition of the leaves of C. villosus

Minerals (mg/kg)
B Ag Ca P Al Pb Cu V Zn Si
0.552 0.0780 234 48.9 6.07 0.008 0.710 0.060 0.654 12.9
Co K Ti Fe Cr Mg Na Mn Ni Sn
0.085 1130 0.002 1.75 0.194 90.0 2.26 1.44 0.239 0.065

3.2 TFC, TPC, and PC contents

The results, presented in Figure 1, indicate that the aqueous extracts of C. villosus have greater concentrations of TFC and PC (337.12 ± 1.2 mg GAE/g and 183.88 ± 2.97 mg EC/g E, respectively) when compared to hydroethanolic extracts. Furthermore, the hydroethanolic extract has a greater flavonoid content (46.33 ± 0.45 mg EQ/g E) than the aqueous extract. Our findings are in accordance with the preceding investigations that showed the abundance of phenols in C. villosus [13,32]. Larit et al. [14] recorded the aerial part of C. villosus, which contained 363.0 ± 8.32 mg GAE/g E phenols and 21.16 ± 1.022 mg QE/g E flavonoids from the n-butanol extract.

Figure 1 
                  PC, TFC, and TPC contents of aqueous and hydroethanolic extracts of C. villosus leaves (mean ± SE).
Figure 1

PC, TFC, and TPC contents of aqueous and hydroethanolic extracts of C. villosus leaves (mean ± SE).

3.3 Phenolic compound quantification in the aqueous extract of C. villosus

The aqueous extract of C. villosus contained 32 polyphenolic compounds (Table 3). The identified chemicals contained polyphenolics including phenolic acids, cinnamic acids, and flavonoids. The major phenolic components identified (Table 3) are (mg/kg) quinic acid (57.478 ± 1.72), salicylic acid (17.38 ± 0.21), isoquercetin (16.895 ± 1.01), gallic acid (15.914 ± 1.51), protocatechuic acid (12.935 ± 0.40), ferulic acid (11.544 ± 0.25), p-coumaric acid (8.873 ± 0.81), vanillic acid (5.377 ± 0.56), gentisic (4.021 ± 0.31), cynaroside (3.145 ± 0.18), caffeic acid (2.711 ± 0.04), quercetin (2.197 ± 0.01), astragalin (1.811 ± 0.61), 4-OH-phenylacetic acid (1.635 ± 0.31), quercetin (1.545 ± 0.55), aromadendrin (1.334 ± 0.09), 3,4,5-trimethoxycinnamic acid (0.978 ± 0.06), and avicularin (0.893 ± 0.17). A few other phenolic compounds were also identified at lower concentrations and some of them were hardly detectable, such as luteolin, rutin, sinapinic acid, kaempferol, naringenin, isorhamnetin, taxifolin, quercetin-3-O-glucuronide, apigenin, hydroferulic acid, pyrocatechol, phloridzin, and resveratrol (Figure 2).

Table 3

Phenolic components identified in the aqueous extract of C. villosus (n = 3)

Class Compounds Molecular formula PubChem number Mass (mg/kg)
Simple phenols Pyrocatechol C6H6O2 289 0.128 ± 0.074
Phenolic acids Gallic acid C7H6O5 370 15.914 ± 1.51
Protocatechuic acid C7H6O4 72 12.935 ± 0.40
Vanillic acid C8H8O4 8468 5.377 ± 0.56
Gentisic acid C7H6O4 3469 4.021 ± 0.31
Salicylic acid C7H6O3 338 17.38 ± 0.21
Phenylacetic acids 4-OH-Phenylacetic acid C8H8O3 127 1.635 ± 0.31
Quinate precursors Quinic acid C7H12O6 6508 57.478 ± 1.72
Hydroxycinnamic acid derivatives p-Coumaric acid C9H8O3 637542 8.873 ± 0.81
Caffeic acid C9H8O4 689043 2.711 ± 0.04
Sinapinic acid C11H12O5 637775 0.73 ± 0.41
Sinapic acid methyl ether C12H14O5 735755 0.978 ± 0.06
Ferulic acid C10H10O4 445858 11.544 ± 0.25
Hydroferulic acid C10H12O4 17865499 0.247 ± 0.049
Flavonoids Isoquercetin C21H20O12 5280804 16.895 ± 1.01
Cynaroside C21H20O11 5280637 3.145 ± 0.18
Astragalin C21H20O11 5282102 1.811 ± 0.61
Quercetin C21H20O11 5280459 1.545 ± 0.55
Aromadendrin C15H12O6 122850 1.334 ± 0.09
Avicularin C20H18O11 5490064 0.893 ± 0.17
Luteolin C15H10O6 5280445 0.765 ± 0.09
Rutin C27H30O16 5280805 0.764 ± 0.01
Kaempferol C15H10O6 5280863 0.598 ± 0.11
Naringenin C15H12O5 439246 0.512 ± 0.05
Isorhamnetin C16H12O7 5281654 0.479 ± 0.140
Taxifolin C15H12O7 471 0.423 ± 0.78
Quercetin-3-Oglucuronide C21H18O13 5274585 0.32 ± 0.012
Apigetrin C21H20O10 5280704 0.642 ± 0,07
Apigenin C15H10O5 5280443 0.27 ± 0.098
Phloridzin C21H24O10 6072 0.052 ± 0.001
Quercetin C15H10O7 5280343 2.197 ± 0.01
Isoflavonoids Daidzin C21H20O9 107971 nd
Stilbenes Resveratrol C14H12O3 445154 0.028 ± 0.001

nd: not determined.

Figure 2 
                  Structures of the main polyphenols detected in the aqueous extract of C. villosus.
Figure 2

Structures of the main polyphenols detected in the aqueous extract of C. villosus.

In the aqueous extract of C. villosus from Algeria, 21 phenolic compounds were identified by Bouziane et al. [12], including apigenin-C-hexoside, quercetin-3-O-glucoside, myricetin-3-O-glucoside myricetin-3-O-rutinoside, myricetin-O-rhamnoside, kaempferol-3-O-rutinoside, myricetin-O-coumaroylrutinoside, and quercetin-O-rhamnoside. The richness and diversity of the polyphenol content of C. villosus offer it many therapeutic possibilities. Notably, ferulic acid has anti-inflammatory and antioxidant capacities [33], and it seems to protect against cardiovascular [34] and renal diseases [35]. Quercetin has been known as an antiviral, antiulcer, anti-inflammatory, and antihypertensive agent [36,37].

3.4 Antioxidant activity

In the current investigation, the PPM assay was used to evaluate the TAC, whereas ABTS, DPPH and reducing power (RP) assays were used to assess the antioxidant activity (Table 4). The molybdate test indicated that the hydroethanolic extract had a high TAC value of 101.14 ± 3.02 mg EAA/g extract, while the aqueous extract showed an antioxidant capacity of 94.18 ± 2.31 mg EAA/g extract. For the DPPH method, the aqueous extract had the most important antioxidant power, with an IC50 (3.94 ± 0.09 μg/mL) compared with the hydroethanolic extract and BHT (IC50 = 4.81 ± 0.061 and 4.15 ± 0.19 μg/mL, respectively). Importantly, the ABTS test indicated that the aqueous extract had a higher level of free radical scavenging activity (IC50 = 2.88 ± 0.07 μg/mL), while the hydroethanolic extract had a lower level, with an IC50 value of 3.32 ± 0.12 μg/mL. These results were equivalent to the activities of ascorbic acid (2.14 ± 0.07 μg/mL).

Table 4

Antioxidant activities (ABTS, DPPH, RP, and molybdate) of C. villosus extracts (mean ± SEM)

IC50 (μg/mL)
Aqueous extracts Hydroethanolic extracts BHT Ascorbic acid
DPPH 3.94 ± 0.09 4.81 ± 0.061* 4.15 ± 0.19
ABTS 2.88 ± 0.07* 3.32 ± 0.12** 2.14 ± 0.07
RP 1.94 ± 0.10* 2.69 ± 0.06** 2.23 ± 0.06
Molybdate (mg AAE/g E) 94.18 ± 2.31 101.14 ± 3.02

* P < 0.05; ** P < 0.01

The RP method confirmed that the effect of the aqueous extract is dose-dependent and higher than that of the hydroethanolic extract and ascorbic acid, with IC50 values of 1.94 ± 0.10 and 2.23 ± 0.06 μg/mL, respectively.

The abundance of phenolic molecules with strong antioxidant activity in C. villosus extracts justifies these results [38]. According to the literature, our results are superior to those reported by Bouziane et al. [12] where it was reported that the aqueous extract of C. villosus from Algeria exhibited IC50 values of 59 ± 2 and 468 ± 34 μg/mL for the DPPH and ABTS tests, respectively. Another study by Aourahoun et al. [39] performed on C. villosus leaves recorded an IC50 of 19.17 μg/mL for the hydroalcoholic extract in the DPPH assay.

The antioxidant activity of the plant studied in this research may be attributed to the correlation between the antioxidant tests and potassium activation in the enzymes that promote the biosynthesis of flavonoids and phenolic compounds [40], which is the most abundant mineral in the leaves of C. villosus. Additionally, common antispasmodic, antimicrobial, anti-inflammatory, and antioxidant properties have been associated with plant metabolites such as proanthocyanidins and flavonoids [41]. Isoquercetin was the most potent DPPH radical scavenger of the quercetin derivatives [42]. Similarly, Yasuda and co-authors [43] proved the DPPH radical scavenging abilities of gallic acid, the key structure of hydrolyzable tannins.

3.5 Anti-inflammatory effect

Our findings (Figure 3) indicated that the inhibition percentages of the aqueous extract of C. villosus (for 500 mg/kg bw) were 42.85 ± 1.27, 57. 95 ± 1.28, and 77.49 ± 0.59% after 1, 3, and 6 h of carrageenan injection, respectively. This effect is remarkable to that of indomethacin (10 mg/kg bw), which is 47.72 ± 0.58, 58.89 ± 0.42, and 75.29 ± 1.07%. Our results show better inhibition percentages relative to results reported by Aourahoun et al. [39] for the hydroethanolic extract of Cytisus triflorus at doses of 200 and 400 mg/kg bw, which exhibited inhibition percentages of 44.19 and 60.50%, respectively. However, our findings are lower than those previously published by Madoui [44], where the inhibition percentages were 80.05 and 88.56% at 4 and 6 h, respectively, for the 400 mg/kg dose of the crude extract of Cytisus triflorus, as the first phase of inflammation is mediated by bradykinin, serotonin, and histamine, whereas the second phase is mediated by prostaglandins and other cytokines [45,46]. The paw edema, which was caused by the accumulation of leukocytes near the inflammatory site, was significantly suppressed by the aqueous extract of C. villosus, a very rich extract in polyphenols. This is exemplified by luteolin derivatives, which protect against GalN/LPS-induced hepatotoxicity through the regulation of inflammatory mediators and phase II enzymes [47]. Moreover, the essential phenolic molecules that we discovered in the aqueous extract of C. villosus such as quercetin, sinapic acid, ferulic acid, caffeic acid, and gallic acid possess effective anti-inflammatory effects [48,49,50,51,52,53]. A study on testicular tissue of young rats by Saygin et al. [54] indicated a strong anti-inflammatory effect of gallic acid by reducing the formation of PGE 2 and calcitonin, and also decreases the expression of IL-6, TNF-α, and TGF-β in rats [48]. In the in vivo model, caffeic acid decreases PGE2 and NO formation and also decreases COX-2, iNOS, and TNF-α expressions through reverse regulation of NF-κB transcription [55]. Using another inflammatory model, Doss et al. revealed that ferulic acid exhibited very interesting anti-inflammatory properties [56]. Lee reported that sinapic acid reduces TNF-α in TNBS-induced colonic inflammation in mice [57]. Importantly, Lesjak et al. [58] found that quercetin reduced the effects of 12-LOX and COX-1, the enzymes involved in the inflammatory response, in a measure-dependent manner.

Figure 3 
                  Anti-inflammatory effects of aqueous extracts of C. villosus. Mean (n = 6) ± SE; **P < 0.001.
Figure 3

Anti-inflammatory effects of aqueous extracts of C. villosus. Mean (n = 6) ± SE; **P < 0.001.

3.6 Inhibition of albumin denaturation

The results of anti-inflammatory activity tests, which were conducted using albumin denaturation, are displayed in Figure 4. According to the results, the aqueous extract has a lower inhibition than the positive control diclofenac sodium (P < 0.0001) with an IC50 = 383.94 and 174.96 μg/mL, respectively.

Figure 4 
                  IC50 values of the inhibition of albumin denaturation of C. villosus and diclofenac sodium. Mean (n = 3) ± SE. The results are considered significantly different for ****
                     P < 0.0001.
Figure 4

IC50 values of the inhibition of albumin denaturation of C. villosus and diclofenac sodium. Mean (n = 3) ± SE. The results are considered significantly different for **** P < 0.0001.

The phytochemical analyses performed indicate that the aqueous extract shows significant amounts of condensed tannins and flavonoids, which may be responsible for this effect. Numerous research studies have demonstrated the well-known and interesting anti-inflammatory properties of flavonoids and condensed tannins [59,60]. The study by Sadique et al. [61] proved that flavonoids trigger STAT-1 and NF-kβ factor by blocking the signal transducer, which would inhibit inflammation.

3.7 Analgesic effects

The peripheral anti-nociceptive performance in rats was determined using the acetic acid-induced torsion method. This approach is widely recognized as a model of visceral inflammatory pain that allows a rapid assessment of this type of analgesic activity. In this method, the release of a number of inflammatory mediators, including cytokines, serotonin, and histamine, is induced by administering acetic acid [62]. Oral dosing of C. villosus (500 mg/kg bw) and aspirin (150 mg/kg bw) exhibited both analgesic activities, with 44.54 and 37.73% contortion inhibition, respectively (Figure 5).

Figure 5 
                  Efficacy of the C. villosus aqueous extract on acetic acid-induced writhing in rats. Mean (n = 6) ± SE; **
                     P < 0.01.
Figure 5

Efficacy of the C. villosus aqueous extract on acetic acid-induced writhing in rats. Mean (n = 6) ± SE; ** P < 0.01.

The central analgesic actions of the aqueous extract and aspirin were assessed using tail-flick and plantar methods. The results are presented in Figures 6 and 7, respectively, and compared with aspirin. In the tail flick method, the aqueous extract (200 and 500 mg/kg) induced significant (P < 0.05) analgesic activity, after 120 min. The highest reaction times for the aqueous extract (200 and 500 mg/kg)-treated groups were 9.13 and 10.46 s, respectively, at 120 min, whereas it was 4.1 and 11.33 s for the saline- and aspirin-treated groups. Similarly, in the plantar method, oral application of an aqueous extract of C. villosus (250, 500 mg/kg) induced significant analgesic activity in a dose-related manner. A peak analgesic activity was reached within 1 h. The pain tolerance times were 4.25 s (P < 0.05) and 5.92 s (P < 0.01) for the aqueous extract at 200 and 500 mg/kg, respectively, whereas it was 7.55 s (P < 0.0001) in the case of aspirin.

Figure 6 
                  Latency of the tail flick of C. villosus. The results are mean of six experiments ± SEM. *
                     P < 0.05.
Figure 6

Latency of the tail flick of C. villosus. The results are mean of six experiments ± SEM. * P < 0.05.

Figure 7 
                  Effect of C. villosus aqueous extract in the plantar test. Mean (n = 6) ± SEM. *
                     P < 0.05.
Figure 7

Effect of C. villosus aqueous extract in the plantar test. Mean (n = 6) ± SEM. * P < 0.05.

The aqueous extract of C. villosus may contain phytochemicals that can block the cyclooxygenase enzyme to diminish pain at the peripheral level or that can intervene on opioid receptors at the central level to reduce this pain. The UPLC analysis of the aqueous extract of our plant shows the presence of analgesic molecules such as quercetin, ferulic acid, quinic acid, and gallic acid that have shown strong analgesic properties in vivo [63,65,66,67]. In the acetic acid-induced pain assay, quercetin (100–500 mg/kg, po) reduced the nociceptive defense response in a drug-related manner [67]. Additionally, quercetin reduced the pain caused by formalin in both phases and reduced the pain caused by capsaicin and glutamate by 75.5 and 68.2%, respectively [67]. Notably, Zhao et al. [63] showed that quercetin reduces the sensation of pain in the feet of mice and improves pain sensitivity, thus showing its analgesic properties. Moreover, ferulic acid increases the nociceptive threshold at doses of 40 and 80 mg/kg in the thermal hyperalgesia test [64]. Di-caffeoyl quinic acids from Lychnophora ericoides showed considerable analgesic action in the acetic acid-induced contortion assay [65]. In addition, da Silva et al. [66] revealed that gallic acid isolated from the leaves of Calophyllum brasiliense had a notable analgesic effect in the contortion test in mice. The results obtained above proved that the aqueous extract of Cytisus villosus has anti-inflammatory and analgesic properties, which confirms the fact that the traditional application of this plant could treat various diseases associated with inflammatory pain.

4 Conclusion

This study highlights the anti-inflammatory, analgesic, and antioxidant abilities of the aqueous extract of the phytodrug Cytisus villosus realized by in vivo and by in vitro tests. Furthermore, UPLC analysis revealed the content of several phenolic molecules with known anti-inflammatory and analgesic effects. These findings indicate that the aqueous extract has the potential to be used in developing a novel analgesic and anti-inflammatory functional food ingredient derived from natural products. Additional investigation is required, however, to confirm these activities by determining the action and toxicities on non-target organisms.


This work was funded by the Researchers Supporting Project (number: RSP-2023R437), King Saud University, Riyadh, Saudi Arabia, and the authors are grateful for the support.

  1. Funding information: This work was funded by the Researchers Supporting Project (No. RSP-2023R437) King Saud University, Riyadh, Saudi Arabia.

  2. Author contributions: Conceptualization, A.Z. and N.E.M.; methodology, Y.E.; software, T.B.; validation, R.E.; formal analysis, T.B., H.A.; investigation, A.E.; resources, A.M.S.; data curation, A.Z., H.N.; writing – original draft preparation, F.K., R.E.; writing – review and editing, M.B.; visualization, L.H.; supervision, L.H.; project administration, L.H., A.M.S.; and funding acquisition.; A.M.S. All authors have read and agreed to the published version of the manuscript.

  3. Conflict of interest: The authors declare that there is no conflict of interest.

  4. Ethical approval: The conducted research is not related to either human or animal use.

  5. Data availability statement: All data generated or analyzed during this study are included in this published article.


[1] Furman D, Campisi J, Verdin E, Carrera-Bastos P, Targ S, Franceschi C, et al. Chronic inflammation in the etiology of disease across the life Span. Nat Med. 2019;25:1822–32. 10.1038/s41591-019-0675-0.Search in Google Scholar PubMed PubMed Central

[2] Calle M, Fernandez ML. Inflammation and type 2 diabetes. Diabetes Metab. 2012;38:183–91. 10.1016/j.diabet.2011.11.006.Search in Google Scholar PubMed

[3] Roth SH. Coming to terms with nonsteroidal anti-inflammatory drug gastropathy. Drugs. 2012;72:873–9. 10.2165/11633740-000000000-00000.Search in Google Scholar PubMed

[4] Russo S. Integrated pain management: using omega 3 fatty acids in a naturopathic model. Tech Reg Anesth Pain Manag. 2008;12:105–8. 10.1053/j.trap.2008.01.001.Search in Google Scholar

[5] Onder G, Pellicciotti F, Gambassi G, Bernabei R. NSAID-related psychiatric adverse events. Drugs. 2004;64:2619–27. 10.2165/00003495-200464230-00001.Search in Google Scholar PubMed

[6] Alhasawi MAI, Aatif M, Muteeb G, Alam MW, Oirdi ME, Farhan M. Curcumin and its derivatives induce apoptosis in human cancer cells by mobilizing and redox cycling genomic copper ions. Molecules. 2022;27:7410. 10.3390/molecules27217410.Search in Google Scholar PubMed PubMed Central

[7] Cristofolini G, Troìa A. A reassessment of the sections of the genus cytisus desf. (Cytiseae, Leguminosae). Taxon. 2006;55:733–46. 10.2307/25065647.Search in Google Scholar

[8] Osórioe, Castro VR. Chromium in a series of Portuguese plants used in the herbal treatment of diabetes. Biol Trace Elem Res. 1998;62:101–6. 10.1007/BF02820025.Search in Google Scholar PubMed

[9] Rivera D, Obón C. The ethnopharmacology of Madeira and Porto Santo Islands, a review. J Ethnopharmacol. 1995;46:73–93. 10.1016/0378-8741(95)01239-A.Search in Google Scholar

[10] Pinela J, Barros L, Carvalho AM, Ferreira ICFR. Influence of the drying method in the antioxidant potential and chemical composition of four shrubby flowering plants from the tribe genisteae (Fabaceae). Food Chem Toxicol. 2011;49:2983–9. 10.1016/j.fct.2011.07.054.Search in Google Scholar PubMed

[11] Larit F, Nael MA, Benyahia S, Radwan MM, León F, Jasicka-Misiak I, et al. Secondary metabolites from the aerial parts of Cytisus villosus pourr. Phytochem Lett. 2018;24:1–5. 10.1016/j.phytol.2017.12.012.Search in Google Scholar PubMed PubMed Central

[12] Bouziane A, Bakchiche B, Dias M, Barros L, Ferreira I, AlSalamat H, et al. Phenolic compounds and bioactivity of Cytisus villosus pourr. Molecules. 2018;23:1994. 10.3390/molecules23081994.Search in Google Scholar PubMed PubMed Central

[13] Benabderrahmane W, Amrani A, Benaissa O, Lores M, Lamas JP, de Miguel T, et al. Chemical constituents, in vitro antioxidant and antimicrobial properties of ethyl acetate extract obtained from Cytisus triflorus l’Her. Nat Product Res. 2020;34:1586–90. 10.1080/14786419.2018.1519816.Search in Google Scholar PubMed

[14] Larit F, León F, Benyahia S, Cutler S. Total phenolic and flavonoid content and biological activities of extracts and isolated compounds of Cytisus villosus pourr. Biomolecules. 2019;9:732. 10.3390/biom9110732.Search in Google Scholar PubMed PubMed Central

[15] Silva LR, Videira R, Monteiro AP, Valentão P, Andrade PB. Honey from Luso Region (Portugal): Physicochemical characteristics and mineral contents. Microchem J. 2009;93:73–7. 10.1016/j.microc.2009.05.005.Search in Google Scholar

[16] Mǎrghitaş LA, Stanciu OG, Dezmirean DS, Bobiş O, Popescu O, Bogdanov S, et al. In vitro antioxidant capacity of honeybee-collected pollen of selected floral origin harvested from Romania. Food Chem. 2009;115:878–83. 10.1016/j.foodchem.2009.01.014.Search in Google Scholar

[17] Amezouar F, Badri W, Hsaine M, Bourhim N, Fougrach H. Évaluation des activités antioxydante et anti-inflammatoire de erica arborea l. Du Maroc. Pathol Biol. 2013;61:254–8. 10.1016/j.patbio.2013.03.005.Search in Google Scholar PubMed

[18] Sayah K, Marmouzi I, Naceiri Mrabti H, Cherrah Y, Faouzi MEA. Antioxidant activity and inhibitory potential of Cistus salviifolius (L.) and Cistus monspeliensis (L.) aerial parts extracts against key enzymes linked to hyperglycemia. BioMed Res Int. 2017;2017:1–7. 10.1155/2017/2789482.Search in Google Scholar PubMed PubMed Central

[19] Zhu Z, Zhang Y, Wang J, Li X, Wang W, Huang Z. Sugaring-out assisted liquid-liquid extraction coupled with high performance liquid chromatography-electrochemical detection for the determination of 17 phenolic compounds in honey. J Chromatogr A. 2019;1601:104–14. 10.1016/J.CHROMA.2019.06.023.Search in Google Scholar

[20] El Omari N, Sayah K, Fettach S, El Blidi O, Bouyahya A, Faouzi ME, et al. Evaluation of in vitro antioxidant and antidiabetic activities of Aristolochia longa extracts. Evid-Based Complement Altern Med. 2019;2019:19. 10.1155/2019/7384735.Search in Google Scholar PubMed PubMed Central

[21] Miguel MG, Nunes S, Dandlen SA, Cavaco AM, Antunes MD. Phenols, flavonoids and antioxidant activity of aqueous and methanolic extracts of propolis (Apis Mellifera L.) from Algarve, South Portugal. Food Sci Technol. 2014;34:16–23. 10.1590/S0101-20612014000100002.Search in Google Scholar

[22] Bougandoura N, Bendimerad N. Evaluation de l'activité antioxydante des extraits aqueux et méthanolique de Satureja calamintha ssp. Nepeta (L.) Briq. Nat Technol. 2013;9:14. in Google Scholar

[23] Council NR. Guide for the care and use of laboratory animals. Vol. 184. Washington, D.C: National Academies Press; 2011.Search in Google Scholar

[24] Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw of the rat as an assay for antiinflammatory drugs. Proc Soc Exp Biol Med. 1962;111:544–7. 10.3181/00379727-111-27849.Search in Google Scholar PubMed

[25] Lekouaghet A, Boutefnouchet A, Bensuici C, Gali L, Ghenaiet K, Tichati L. In vitro evaluation of antioxidant and anti-inflammatory activities of the hydroalcoholic extract and its fractions from Leuzea conifera L. roots. South Afr J Botany. 2020;132(1):103–7.10.1016/j.sajb.2020.03.042Search in Google Scholar

[26] Sayah K, Chemlal L, Marmouzi I, El Jemli M, Cherrah Y, Faouzi MEA. In vivo anti-inflammatory and analgesic activities of Cistus salviifolius (L.) and Cistus monspeliensis (L.) aqueous extracts. South Afr J Bot. 2017;113:160–3. 10.1016/j.sajb.2017.08.015.Search in Google Scholar

[27] Sood S, Arora B, Bansal S, Muthuraman A, Gill NS, Arora R, et al. Antioxidant, anti-inflammatory and analgesic potential of the Citrus decumana l. peel extract. Inflammopharmacology. 2009;17:267–74. 10.1007/S10787-009-0015-Y.Search in Google Scholar PubMed

[28] El Hamsas El Youbi A, El Mansouri L, Boukhira S, Daoudi A, Bousta D. In vivo anti-inflammatory and analgesic effects of aqueous extract of Cistus Ladanifer L. from Morocco. Am J Ther. 2016;23:e1554–9. 10.1097/MJT.0000000000000419.Search in Google Scholar PubMed

[29] Elbadrawy E, Sello A. Evaluation of nutritional value and antioxidant activity of tomato peel extracts. Arab J Chem. 2016;9:S1010–8. 10.1016/j.arabjc.2011.11.011.Search in Google Scholar

[30] Grela ER, Samolińska W, Kiczorowska B, Klebaniuk R, Kiczorowski P. Content of minerals and fatty acids and their correlation with phytochemical compounds and antioxidant activity of leguminous seeds. Biol Trace Elem Res. 2017;180:338–48. 10.1007/s12011-017-1005-3.Search in Google Scholar PubMed PubMed Central

[31] Evans P, Halliwell B. Micronutrients: Oxidant/antioxidant status. Br J Nutr. 2001;85:S67. 10.1049/BJN2000296.Search in Google Scholar

[32] Madoui S, Charef N, Arrar L, Baghianni A, Khennouf S. In vitro antioxidant activities of various extracts from flowers-leaves mixture of Algerian Cytisus Triflorus. Annu Res Rev Biol. 2018;26:1–13. 10.9734/ARRB/2018/41297.Search in Google Scholar

[33] Zduńska K, Dana A, Kolodziejczak A, Rotsztejn H. Antioxidant properties of ferulic acid and its possible application. Skin Pharmacol Physiol. 2018;31:332–6. 10.1159/000491755.Search in Google Scholar PubMed

[34] Neto-Neves EM, da Silva Maia Bezerra Filho C, Dejani NN, de Sousa DP. Ferulic acid and cardiovascular health: therapeutic and preventive potential. Mini-Rev Med Chem. 2021;21:1625–37. 10.2174/1389557521666210105122841.Search in Google Scholar PubMed

[35] Alam MA, Sernia C, Brown L. Ferulic acid improves cardiovascular and kidney structure and function in hypertensive rats. J Cardio Pharm. 2013;61:240–9. 10.1097/FJC.0b013e31827cb600.Search in Google Scholar PubMed

[36] Ullah MF, Bhat S, Hussain E, Abu-Duhier F, Khan HY, Aatif M, Dietary phenolics as cancer chemopreventive nutraceuticals: a promising paradigm. In: Atta-ur-Rahman, Choudhary MI, editors. Frontiers in Anti-Cancer Drug Discovery. Bentham Science Publishers; 2013. p. 32–92.10.2174/9781608058082113020005Search in Google Scholar

[37] Nabavi SF, Russo GL, Daglia M, Nabavi SM. Role of quercetin as an alternative for obesity treatment: You Are What You Eat! Food Chem. 2015;179:305–10. 10.1016/j.foodchem.2015.02.006.Search in Google Scholar PubMed

[38] Sadeq O, Mechchate H, Es-safi I, Bouhrim M, Jawhari FZ, Ouassou H, et al. Phytochemical screening, antioxidant and antibacterial activities of pollen extracts from micromeria fruticosa, achillea fragrantissima, and phoenix dactylifera. Plants. 2021;10:676. 10.3390/plants10040676.Search in Google Scholar PubMed PubMed Central

[39] Aourahoun KA, Fazouane F, Benayache S. Pharmacological potential of cytisus triflorus l’hérit. extracts as antioxidant and anti-inflammatory agent. Der Pharm Lett. 2015;7:104–10.Search in Google Scholar

[40] Tamuly C, Saikia B, Hazarika M, Bora J, Bordoloi MJ, Sahu OP. Correlation between phenolic, flavonoid, and mineral contents with antioxidant activity of underutilized vegetables. Int J Veg Sci. 2013;19:34–44. 10.1080/19315260.2012.671237.Search in Google Scholar

[41] Iqbal E, Salim KA, Lim LBL. Phytochemical screening, total phenolics and antioxidant activities of bark and leaf extracts of goniothalamus velutinus (Airy Shaw) from Brunei Darussalam. J King Saud Univ Sci. 2015;27:224–32. 10.1016/j.jksus.2015.02.003.Search in Google Scholar

[42] Kato K, Ninomiya M, Tanaka K, Koketsu M. Effects of functional groups and sugar composition of quercetin derivatives on their radical scavenging properties. J Nat Prod. 2016;79:1808–14. 10.1021/acs.jnatprod.6b00274.Search in Google Scholar PubMed

[43] Yasuda T, Inaba A, Ohmori M, Endo T, Kubo S, Ohsawa K. Urinary metabolites of gallic acid in rats and their radical-scavenging effects on 1,1-diphenyl-2-picrylhydrazyl radical. J Nat Prod. 2000;63:1444–6. 10.1021/np0000421.Search in Google Scholar PubMed

[44] Madoui S. Activités biologiques des extraits de Cytisus Triflorus [dissertation]. Université Ferhat Abbas - Sétif 1; 2018.Search in Google Scholar

[45] Guay J, Bateman K, Gordon R, Mancini J, Riendeau D. Carrageenan-induced paw edema in rat elicits a predominant prostaglandin E2 (PGE2) response in the central nervous system associated with the induction of microsomal PGE2 synthase-1. J Biol Chem. 2004;279:24866–72. 10.1074/jbc.M403106200.Search in Google Scholar PubMed

[46] Roy S, Khanna S, Shah H, Rink C, Phillips C, Preuss H, et al. Human genome screen to identify the genetic basis of the anti-inflammatory effects of boswellia in microvascular endothelial cells. DNA Cell Biol. 2005;24:244–55. 10.1089/dna.2005.24.244.Search in Google Scholar PubMed

[47] Park CM, Song YS. Luteolin and luteolin-7-O-glucoside protect against acute liver injury through regulation of inflammatory mediators and antioxidative enzymes in GalN/LPS-induced hepatitic ICR mice. Nutr Res Pract. 2019;13:473–9. 10.4162/nrp.2019.13.6.473.Search in Google Scholar PubMed PubMed Central

[48] Wei G, Wu Y, Gao Q, Shen C, Chen Z, Wang K, et al. Gallic acid attenuates postoperative intra-abdominal adhesion by inhibiting inflammatory reaction in a rat model. Med Sci Monit. 2018;24:827–38. 10.12659/MSM.908550.Search in Google Scholar PubMed PubMed Central

[49] Dludla P, Nkambule B, Jack B, Mkandla Z, Mutize T, Silvestri S, et al. Inflammation and oxidative stress in an obese state and the protective effects of gallic acid. Nutrients. 2018;11:23. 10.3390/nu11010023.Search in Google Scholar PubMed PubMed Central

[50] Chao P, Hsu C, Yin M. Anti-inflammatory and anti-coagulatory activities of caffeic acid and ellagic acid in cardiac tissue of diabetic mice. Nutr Metab. 2009;6:33. 10.1186/1743-7075-6-33.Search in Google Scholar PubMed PubMed Central

[51] Cao YJ, Zhang Y, Qi JP, Liu R, Zhang H, He LC. Ferulic acid inhibits H2O2-induced oxidative stress and inflammation in rat vascular smooth muscle cells via inhibition of the NADPH oxidase and NF-KB pathway. Int Immunopharmacol. 2015;28:1018–25. 10.1016/j.intimp.2015.07.037.Search in Google Scholar PubMed

[52] Zhang Q, Hu JX, Kui X, Liu C, Zhou H, Jiang X, et al. Sinapic acid derivatives as potential anti-inflammatory agents: synthesis and biological evaluation. Iran J Pharm Res. 2017;16:1405–14.Search in Google Scholar

[53] Kim Y, Park W. Anti‐inflammatory effect of quercetin on RAW 264.7 mouse macrophages induced with polyinosinic‐polycytidylic acid. Molecules. 2016;21:450. 10.3390/molecules21040450.Search in Google Scholar PubMed PubMed Central

[54] Saygin M, Asci H, Ozmen O, Cankara FN, Dincoglu D, Ilhan I. Impact of 2.45 GHz microwave radiation on the testicular inflammatory pathway biomarkers in young rats: the role of gallic acid. Environ Toxicol. 2016;31:1771–84. 10.1002/tox.22179.Search in Google Scholar PubMed

[55] Shin KM, Kim IT, Park YM, Ha J, Choi JW, Park HJ, et al. Anti-inflammatory effect of caffeic acid methyl ester and its mode of action through the inhibition of prostaglandin E2, nitric oxide and tumor necrosis factor-α production. Biochem Pharmacol. 2004;68:2327–36. 10.1016/j.bcp.2004.08.002.Search in Google Scholar PubMed

[56] Doss HM, Dey C, Sudandiradoss C, Rasool MK. Targeting inflammatory mediators with ferulic acid, a dietary polyphenol, for the suppression of monosodium urate crystal-induced inflammation in rats. Life Sci. 2016;148:201–10. 10.1016/j.lfs.2016.02.004.Search in Google Scholar PubMed

[57] Lee JY. Anti-inflammatory effects of sinapic acid on 2,4,6-trinitrobenzenesulfonic acid-induced colitis in mice. Arch Pharmacal Res. 2018;41:243–50. 10.1007/s12272-018-1006-6.Search in Google Scholar PubMed

[58] Lesjak M, Beara I, Simin N, Pintać D, Majkić T, Bekvalac K, et al. Antioxidant and anti-inflammatory activities of quercetin and its derivatives. J Funct Foods. 2018;40:68–75. 10.1016/j.jff.2017.10.047.Search in Google Scholar

[59] Manach C, Williamson G, Morand C, Scalbert A, Rémésy C. Bioavailability and bioefficacy of polyphenols in humans. i. review of 97 bioavailability studies. Am J Clin Nutr. 2005;81:230S–42S. 10.1093/ajcn/81.1.230S.Search in Google Scholar PubMed

[60] Ayinke A, Morakinyo M, Olalekan I, Philip T, Mariam O, Oluokun O. In vitro evaluation of membrane stabilizing potential of selected bryophyte species. Eur J Med Plants. 2015;6:181–90. 10.9734/EJMP/2015/7629.Search in Google Scholar

[61] Sadique J, Chandra T, Thenmozhi V, Elango V. Biochemical modes of action of cassia occidentalis and cardiospermum halicacabum in inflammation. J Ethnopharmacol. 1987;19:201–12. 10.1016/0378-8741(87)90042-0.Search in Google Scholar PubMed

[62] Deraedt R, Jouquey S, Delevallée F, Flahaut M. Release of prostaglandins E and F in an algogenic reaction and its inhibition. Eur J Pharmacol. 1980;61:17–24. 10.1016/0014-2999(80)90377-5.Search in Google Scholar PubMed

[63] Zhao ZY, Guo L, Wang XB, Han SY. Anti-inflammatory and analgesic effects of quercetin chitosan composite solution. Chin J Tissue Eng Res. 2013;16:8803–6. 10.3969/j.issn.2095-4344.2012.47.014.Search in Google Scholar

[64] Lv WH, Zhang L, Wu SJ, Chen SZ, Zhu XB, Pan JC. Analgesic effect of ferulic acid on CCI mice: behavior and neurobiological analysis. Zhongguo Zhong Yao Za Zhi. 2013;38:3736–41.Search in Google Scholar

[65] Dos Santos MD, Gobbo-Neto L, Albarella L, Petto De Souza GE, Lopes NP. Analgesic activity of di-caffeoylquinic acids from roots of lychnophora ericoides (Arnica Da Serra). J Ethnopharmacol. 2005;96:545–9. 10.1016/J.JEP.2004.09.043.Search in Google Scholar

[66] da Silva KL, dos Santos AR, Mattos PE, Yunes RA, Delle-Monache F, Cechinel-Filho V. Chemical composition and analgesic activity of calophyllum brasiliense leaves. Therapie. 2001;56:431–4.Search in Google Scholar

[67] Filho AW, Filho VC, Olinger L, De Souza MM. Quercetin: further investigation of its antinociceptive properties and mechanisms of action. Arch Pharmacal Res. 2008;31:713–21. 10.1007/S12272-001-1217-2.Search in Google Scholar

Received: 2023-02-13
Revised: 2023-04-23
Accepted: 2023-05-05
Published Online: 2023-06-14

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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