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The effects of the aqueous crude extract (5%) of Eugenia punicifolia on cholinergic nicotinic neurotransmission were investigated. Actions of. Eugenia punicifolia (Kunth) DC (Myrtaceae), popularly known as pedra-ume-caá, pedra-ume, murta, or muta, is a shrub found mainly in the. Figure 16 Body weight evolution in rats with acetic acid-induced gastric ulcers treated with oral administration of vehicle, lansoprazole.

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To evaluate the sex-specific effects of a hydroalcoholic extract from Eugenia punicifolia HEEP leaves on gastric ulcer healing. In this rat study involving males, intact cycling females, and ovariectomized females, gastric ulcers were induced using acetic acid. A vehicle, lansoprazole, or HEEP was administered for 14 d after ulcer induction.

Body weight was monitored throughout the treatment period. At the end of treatment, the rats were euthanized and the following in vivo and in vitro investigations were performed: Additionally, the fugenia effect of HEEP was evaluated in terms of subacute toxicity and cytotoxicity. Compared to the vehicle, HEEP demonstrated a great healing capacity by substantially reducing the ulcerative lesion area in males Overall, the beneficial effect of HEEP was the highest in intact females.

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Notably, HEEP promoted the expression of vascular endothelial growth factor intact vs ovariectomized females and decreased the expression of Caspase-8 and Bcl-2 intact female vs male or ovariectomized female.

Additionally, HEEP enhanced fibroblast proliferation and migration into a wounded area in vitroconfirming its healing effect. Finally, no sign of punciifolia toxicity or cytotoxicity of HEEP punicifoolia observed.

In gastric ulcers, HEEP-induced healing modulated by female sex hormones; in males, mediated by prostaglandin involves extracellular matrix remodeling, with gastric mucosa cell proliferation and migration.

Gastric ulcers, which occur due to an imbalance between protective and aggressive agents at the gastric pjnicifolia surface, is a chronic disease that affects millions worldwide and has high rates of relapse.

The conventional treatment for gastric ulcers is associated with several side effects and poor healing of the gastric mucosa. Eugenia punicifolia is a euvenia plant used to treat inflammation and wounds. The present study in rats with gastric ulcers confirms the healing effect of Eugenia punicifolia extract and clarifies its differential effect in males and females.

These findings are useful for developing novel and safe therapies for gastric ulcers.

Eugenia punicifolia (Kunth) DC.

Gastric ulcers, which result from an imbalance between the protective and aggressive agents at the surface of the gastric mucosa, is a chronic disease that affects millions around the world punicifolua has high relapse rates[ 12 ].

Alcohol consumption, prolonged treatment with non-steroidal punucifolia drugs, stress, and Helicobacter pylori infection favor such an imbalance and represent key etiological factors of gastric ulcers[ 3 ].

Current treatment strategies for gastric ulcers involve the use of anti-secretory drugs, including antagonists of histamine receptor type 2 e. While such treatments are effective, prolonged use of anti-secretory drugs, especially ranitidine and proton pump inhibitors, is associated with several side effects[ 4 ] and poor healing of the gastric mucosa[ 5 ].

InTarnawski et al[ 6 ] proposed the concept of quality of ulcer healing, which takes into consideration the fact that tissue regeneration within the ulcer scar is often incomplete.

Within the quality of ulcer healing concept, the evaluation of gastric ulcer healing is focused on whether the structure and function of the mucosal and submucosal tissue have recovered completely, in addition to endoscopic examination and evaluation of ulcer size.

It has been shown that ulcer recurrence is closely related to the quality of ulcer healing[ 7 ]. Ulcers frequently recur following treatment with anti-secretory drugs[ 78 ]. Therefore, alternative therapies for gastric ulcers are desirable[ 9 ].


Herbal combination preparations are popular among traditional herbal medicine practitioners. The rationale behind such combinations is frequently questioned, and it remains challenging to assess the individual contribution of each component to the overall activity of the herbal combination preparation. This holds especially true when the preparation is used in the treatment of a chronic multifactorial disease such as gastric ulcers[ 10 ].

The leaves of E. The gastroprotective activity of the hydroalcoholic extract of E. Nevertheless, it remains unclear whether HEEP has any beneficial effects in the healing of installed gastric ulcers, since the gastroprotective activities of an extract do not ensure their gastric healing effects in installed gastric ulcers[ 17 ].

Therefore, the present study aimed to evaluate the sex-specific effects of HEEP in the healing of gastric ulcers in a rat model. For this purpose, we employed a rat model of acetic acid-induced gastric ulcers and analyzed the curative action of HEEP in males, intact females, and ovariectomized females.

The following chemicals and reagents were used: The same saline solution used as a vehicle served as a negative control. In DecemberDr. Catarina dos Santos collected the leaves of E. The dried and crushed leaves 10 g of plant material were dissolved in mL of solvent consisting of a Thereafter, the solution was filtered, and the residue was extracted twice more.

The animal protocol was designed to minimize pain or discomfort to the animals. The in vivo experiments used male g and female g Wistar albino rats obtained from the breeding facility of the State University of Campinas Multidisciplinary Center for Biological Research. The animals were kept in cages with raised, wide-mesh floors to prevent coprophagy. The estrous cycle was verified through a vaginal smear performed daily starting on postnatal day The material was observed under an optical microscope, and the estrous cycle phase was determined by cytology[ 18 – 20 ].

The duration of the estrous cycle was calculated as the number of days between one estrus phase and the next. Only female rats that showed two consecutive regular cycles of d were included in the experiment. Additionally, ovariectomized OVZ females were included.

We performed bilateral ovariectomies in ten-week-old female rats and we verified the absence punicifoliaa estrous cycle through a vaginal smear after 2 wk. The experiments were performed in 90 d old males as well as in intact and OVZ female rats.

Eugenia punicifolia – Wikipedia

The 84 animals included in the study 30 males, 30 intact females, 24 ovariectomized females were divided into nine groups with individuals each. Group allocation was performed according to sex male vs femalehormonal status in females intact vs OVZand treatment negative control vs positive control vs HEEP. All subsequent analyses except for the cell viability and migration assays were performed in each of the nine groups. A plastic tube with an internal diameter of 4. The acetic acid was left to act for 20 s and then completely removed.

The stomach was bathed with saline to avoid adherence to the external surface of the ulcerated region, and the abdomen was closed. Subsequently, all animals were fed normally. In order to determine the healing effect of HEEP, three d treatment protocols were evaluated in this study. The three groups of negative controls males, intact females, and OVZ females received 0. All treatments were delivered orally once daily beginning one day after surgery and continuing for 14 d. One day after the last drug administration, the rats were euthanized through decapitation and the stomachs were removed to evaluate the lesions.


The ulcer tissue was also removed and analyzed. The organs were removed and weighed. Additionally, analysis of blood collected upon euthanasia was performed. The animals were not anesthetized prior to decapitation, since anesthetics may interfere with the results of the biochemical parameters evaluated. The percentage of viable cells was calculated as Abs. The proliferation ability of fibroblasts exposed to HEEP was assessed using the scratch-wound assay, according to a protocol previously described by Balekar et al[ 24 ], with a few modifications.

The scratch test measures the expansion eugenai a cell population on surfaces. Any cellular debris created while making the scratch was removed by gently washing the wells with PBS. The plates were evaluated after 24 h and 48 h of incubation to assess the closure of the scratch wounds. All experiments were performed in triplicate.

Tissue containing the gastric ulcer was used to extract total protein. Finally, the protein content was quantified using the biuret method[ 2526 ]. The complete protocol for gelatin zymography was previously punicifoloa by Justulin et al[ 27 ]. After electrophoresis, the gels were subjected to two 15 min washes in a solution of 2.

Finally, the puniciffolia were stained with Coomassie Brilliant Blue R The bands obtained through zymography were scanned and analyzed by densitometry.

Eugenia calycina and punicifolia

The bands representative of the gelatinase activity of matrix metalloproteinase MMP -2 and MMP-9 were analyzed in terms of the integrated optical density IOD of the bands.

For this test, the protein extracts were obtained as described above eugejia zymography. After extraction, the samples were treated with Laemmili buffer 0. The membranes were washed with PBS and incubated with punicifolua specific secondary antibody for 2 h horseradish peroxidase, 1: After washing, the reactions were detected using an enhanced chemiluminescence kit Amersham Biosciences, Westborough, MA, United States and the punicofolia were captured using a G: The expression levels were normalized to that of actin 1: Body weight was recorded daily throughout the experimental period.

Macroscopic analyses punidifolia weighing of the vital organs liver, kidneys, heart, spleen, and lungs were performed at the end of the treatment. The statistical methods of this study were reviewed by Dr.

To evaluate the HEEP-mediated healing effect of gastric ulcers, we measured the gastric lesion area at the end of treatment. In male rats, d treatment with HEEP was associated with significantly decreased lesion area Among females, HEEP was associated with a very significant reduction in lesion area, both in cycling rats Effects of d treatment with hydroalcoholic extract from the leaves of Eugenia punicifolia and lansoprazole on gastric ulcer healing in a rat model.

Values are given as a percentage indicating healing relative to the corresponding vehicle group. Hydroalcoholic extract from the leaves of Eugenia punicifolia. No cytotoxicity signal was found after 24 h of incubation with HEEP, regardless of concentration 0. Cell viability was measured using the MTT assay. We performed an in vitro scratch-wound test to determine whether HEEP possesses cell proliferation activity. Effects of hydroalcoholic extract from the leaves of Eugenia punicifolia on cell proliferation and migration into the scratch-wound area.

Rate of migration, evaluated in terms of the total distance that the cells moved from the edge of the scratch toward the center. Prostaglandin E 2 levels in rats with acetic acid-induced eugenua ulcers treated for 14 d with vehicle, lansoprazole, or hydroalcoholic extract from the leaves of Eugenia punicifolia. Hydroalcoholic extract from the leaves of Eugenia punicifolia ; PGE 2: