PDF | Libidibia ferrea, popularly known as “pau ferro” or “jucá”, is an important medicinal plant employed for the treatment of various ailments in. Genus: Libidibia (formerly known as Ceasalpinia). species: ferrea. Family: Fabaceae. Common Name: Leopard tree, Brazilian Ironwood. Description. Unarmed tree, to 15 m. Bark very distinctive, smooth, grey, pealing to reveal paler patches. Leaves 2-pinnate; pinnae in pairs, each with

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To receive news and publication updates for BioMed Research International, enter your email address in the box below. This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Libidibia ferrea LF is a medicinal plant that holds many pharmacological properties. Histochemical tests and Gas chromatography of the LPLF were performed to characterize its fatty acids.

Acetic acid-induced abdominal constriction, formalin-induced pain, and hot-plate test in mice were employed in the study. LF aqueous seed extract and LPLF demonstrated a dose-dependent antinociceptive effect in all tests indicating both peripheral anti-inflammatory and central analgesia properties.

Libidibia ferrea

frerea Together, the present results suggested that Libidibia ferrea induced antinociceptive activity is possibly related to its ability to inhibit opioid, cholinergic receptors, and cyclooxygenase-2 pathway, since its main component, linoleic acid, has been demonstrated to produce such effect in previous studies.

Libidibia ferrea LF is a plant used in folk medicine for therapeutical purposes. LF is a tree species from the Leguminosae family, one of the largest dicotyledon families with approximately genus, that gathers more than 18, species.

It is found along all tropical regions in Brazil but manly in the north and northeast areas [ 1 ]. Previous studies identified many pharmacological properties of LF, which explain its common use in folk medicine, for treatment of wounds, bruises, chronic cough, and asthma [ 2 ]. The fruits can be used to treat diabetes and to prevent cancer. Its extract proved in in vivo tests wound healing [ 3 ]. Roots are utilized as antipyretic and treatment of diarrhea. LF has also demonstrated anticancerogenic, antioxidant, wound repair, and DNA protection properties [ 45 ].

Besides, the bark is used for enterocolitis, diabetes, and rheumatism treatment [ 6 ]. Thus, in the present study, we identified the main storage substances and their accumulation sites in Libidibia ferrea seeds, identified its main lipid components, and investigated the analgesic effect of LF on chemical and thermal models of nociception in mice and its possible mechanism of action. The specimen was identified as Libidibia ferrea or their synonym Caesalpinia ferrea.

After collecting the material, seeds of Libidibia ferrea were separated for extraction. For the preparation of the acetonic extract, seeds collected were dried up gerrea natural libidiia, powdered, and put into erlenmeyers.

Then, acetone was added and the solution was incubated at one week. Histochemical tests were carried libidibix using free-hand sections of newly collected material and included the Sudan IV test for lipids, Osmium tetroxide test for unsaturated lipids, and Nile blue sulphate test to neutral and acids lipids.

Anatomical and histochemical descriptions were made with the aid of image analysis software Axiovision using an Axioplan Zeiss microscope. Fatty acid composition was calculated as a percentage of the total fatty acids present in the sample.

All experiments reported in this study were carried out in ferreq with current guidelines for the care of laboratory animals and ethical guidelines for investigation of experimental pain in conscious animals. All efforts were made to minimize the number of animals fereea and their suffering. The present study utilized the following drugs: All drugs were dissolved in saline just before use, except celecoxib, which was dissolved in methylcellulose 0.

LPLF was given orally p. The negative control group was given saline, always in the same route of administration ferre the drug. The doses of all substances used in this work were selected based on previous results cerrea our laboratory [ 8 ]. Abdominal contraction, ferrda by ljbidibia. The animals were pretreated i.

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The control groups received the same volume, 0. Antinociceptive activity was expressed fferrea the reduction in the number of abdominal constrictions. Frrrea procedure used was similar to that described previously [ 10 ]. Twenty microliters of 2. The amount of time spent licking the injected paw was timed with a chronometer and was considered as indicative of nociception. The hot-plate test was used to measure the response latencies according to the method described previously [ 12 ].


The time necessary for the initial response to the painful stimulus taping of the paws, licking or jumping was taken as defining nociceptive response.

The nociceptive response was evaluated by the acetic acid-induced contortions and the evaluation of the mechanism of action was determined by the reversion of the LF seed LPLF antinociceptive effect. The ID 50 values were calculated from at least three dosages of the extract and LPLF, determined by linear regression from individual experiments using appropriate software SigmaPlot software, version libbidibia Libidibia ferrea is a known popular used plant for treatment of different pain related injuries.

In order to identify the possible analgesic effect of the LF aqueous seed extract, an abdominal writhing test was performed. Therefore, LF aqueous extract demonstrated an antinociceptive effect in chemical induced peripheral pain Figure 1.

The plant seeds are one of the tree parts used for the analgesic treatment. So Histochemical tests of the seed were performed to identify its components. Libidibia ferrea presented unitegumented seed composed of exotest epidermis with a thin cuticle layer; a subcuticular space and a palisade layer were formed by macrosclereids. A mesotest composed of osteosclereids and several layers of fibers, and an endotest composed of an inner surface; the space between the exotest and the cotyledon was occupied for endosperm Figure 2 a.

In the cotyledon, a great amount of the lipids were revealed with the Sudan IV test Figure 2 b and a thin cuticle layer in the exotest epidermis Figure 2 c. Osmium tetroxide test revealed that most of the totality of lipids observed was mainly unsaturated and predominantly observed in the cotyledon Figure 2 din the macrosclereids, and in the thicker cell walls of the fibers Figures 2 e – 2 f. Additionally, Nile blue sulphate test reveals the presence of the neutral and acid lipids in the cuticle and its cotyledonary adaxial epidermis and Figure 2 g and mainly in the subcuticular space Figures 2 h – 2 i.

Terpenoids were also observed only in the cotyledonary adaxial surface through the NADI reagent test Figure 2 j. In summary, Libidibia ferrea has a large amount of lipids storage sites in different seed parts containing different lipids unsaturated, neutral, and acid.

The percentages of the obtained values of GC for the chemical LPLF composition indicated a prevalence of unsaturated fatty libidibai as demonstrated in Table 1.

LPLF could cause pain lbiidibia mainly by two libiddibia pain mechanisms: To investigate the analgesic effect in these different pain types, formalin was injected in the plantar left paw of pretreated mice and the time that they fsrrea licking was counted. The calculated mean ID 50 value for the early phase was 5. As previous data show Libidibia ferrea seed causes inhibition on neurogenic pain and to confirm derrea form of antinociception, a hot-plate test was performed. Pretreated mice were put on a heated plate and the time that took for them to jump, tap their foot, or lick their paw was counted.

The LPLF caused an increase in the latency time of the animals in all analyzed periods at 2. To evaluate by which central pain transmission pathway LPLF was acting on, mice were pretreated with naloxone and atropine 1. Naloxone and atropine were able to reverse LPLF antinociception.

These data indicate that the LPLF might act on both opioid and cholinergic pathways.

Also, we discovered that aqueous extract and LPLF presented antinociceptive effects and that LPLF demonstrated peripheral and central forms of antinociception. First, LF aqueous extract was tested in visceral nociception induced by acetic acid, which demonstrated a significant analgesic effect Figure 1.

This data corroborates with two other studies [ 1314 ]; however, both demonstrated lower analgesic effect. The greater efficiency of the analgesic effect could be due to a greater presence of active compound s in the plant seeds.

Due to a significant layer of oil present in the seed extract used in the experiments, it was pondered if the analgesic agent s were present in this lipophilic portion. So the LPLF was isolated and GC evaluated its components, in order to identify analgesic related components, and histological assessments of the seeds were performed in order to indicate the storage site of such substances. Histochemical seed analyses showed a mesotest composition and storage of endosperm between the exotest and cotyledon Figures 2 b and 2 cwhich is commonly the main seed storage tissues in the endospermic seeds, although LF presented a complete absorption of the endosperm during the embryogenesis [ 715 ].


The lipids found in Sudan IV test Figure 2 b occurred as oily bodies in different tissues of the seeds, such as cotyledon, endosperm, and embryo, since other tissues, beyond the cotyledon, can contribute to the storage of substances in the mature seed [ 1516 ].

Additionally, findings in Nile blue sulphate test possibly confirm the presence of the triacylglycerols in the oily bodies Figures 2 g and 2 h. The presence of terpenoids Figure 2 jessential oil compounds found in seeds, flowers, leaves, roots, and wood of higher plants, could be related to previous detected LF antimicrobial activity, which has already been characterized as an LF property [ 17 ].

To identify the exact components present on the seed oily bodies and to further investigate the source of the antinociceptive effect, a Gas chromatography of the LPLF was performed Figure 3. It identified the following components: In the analyses, linoleic acid This seems to be common in this genus since Caesalpinia sappan [ 18 ] and Caesalpinia echinata [ 19 ] also have linoleic acid as their main fatty acid component with But some other species such as Caesalpinia bonducella has hexadecadienoate According to Mayworm and Salatino [ 21 ] most species from the Fabaceae family have predominance of the oleic acid and low quantities of fatty acids with more than 20 carbons, showing that LF differs from its family main lipid storage forms.

LF presented also a majority of unsaturated fatty acids which are represented as linoleic, oleic, linolenic, and palmitoleic acid, similar to C. In the meantime, C. The saturated fatty acids were found in a minor concentration in LF extract. The principal saturated fatty acid was the palmitic acid and in lower concentrations of capric and stearic acids.

This ferrsa show similarity with C. After these analyses, LPLF was used in subsequent antinociceptive tests libidobia and formalin-induced lickingwhich demonstrated a significant analgesic effect in both tests, confirming that the antinociceptive effect in the seeds extract could be attributed to at least one of LPLF components.

In formalin-induced licking test, LPLF demonstrated inhibition in both early and late phases. Data of the LPLF in the late phase indicate a strong inhibition in peripheral inflammatory induced pain Figure 4 b. Omega-3 fatty acids, mainly linoleic acid, are used, in libidibbia, as a supplementation for inflammatory joint pain [ 2324 ] and plants with high concentrations of linoleic acid usually have also analgesic effect [ 25 ].

Libidibia ferrea – Wikipedia

On the other hand, the same stimuli could be activating MyD88, which activates the Akt pathway. Since anti-inflammatory drugs decrease the concentration of PGE 2which is responsible for activating and lowering the excitatory threshold of peripheral polimodal nociceptive neurons, anti-inflammatory drugs can cause a secondary analgesic effect. It is possible that the libivibia acid might be acting through this mechanism [ 31 ].

These results are more significant from those found by Carvalho et al. First phase of formalin tests predominantly evokes activity in A-delta fibers. Experimental results have indicated that substance P and bradykinin participate in this early phase [ 32 ]. It suggests that LF compounds could be promoting the antinociceptive effect in A-delta fibers. To better characterize what peripheral and central pathways that LPLF was affecting, abdominal writhing test with the administration of different inhibitors was performed Figure 6.

Reversal of the antinociceptive effect by opioid antagonist indicates a participation in central forms of antinociception. However, further pharmacological studies are necessary to confirm this hypothesis.