Prophylactic use of standardized extract of propolis of Apis mellifera (EPP-AF®) reduces lung inflammation and improves survival in experimental lethal sepsis.

ETHNOPHARMACOLOGICAL RELEVANCE: Sepsis poses one of the biggest public health problems, necessitating the search for new therapeutic alternatives. For centuries, propolis has been widely used in folk medicine to treat various inflammatory and infectious diseases. Given its extensive use, it has excellent potential as an adjuvant treatment for patients with sepsis. OBJECTIVE: This study evaluated prophylactic treatment with standardized propolis extract (EPP-AF®) and followed the prognosis of sepsis induced by ligation and cecal ligation and puncture (CLP). METHODS: Initially, for survival assessment, Swiss mice were separated into five groups: Sham (false operated), control (PBS), ATB (received antibiotic, 8 mg/kg), P10 (received EPP-AF®, 10 mg/kg), and P100 (received EPP-AF®, 100 mg/kg). The animals received PBS, antibiotic, or EPP-AF® by the subcutaneous route 6 h before the CLP procedure. Animal survival was assessed every 12 h for five days when all of them were euthanized. RESULTS: We show that the treatment with EPP-AF® significantly increased the life expectancy of animals with sepsis compared to the control group. Interestingly, prophylactic treatment with EPP-AF® showed no effect on the number of colony-forming units in the peritoneum, blood, or lung. However, there was a decrease in cellular influx in the peritoneum. This alteration was unrelated to the number of bone marrow cells or the differential counting of peripheral blood cells. The coagulogram remained unchanged, including the number of platelets and prothrombin time-activated partial thromboplastin time. However, the inflammatory infiltrate and bleeding in the lung tissue were lower in the animals that received EPP-AF®. CONCLUSION: Thus, it was possible to conclude that prophylactic treatment with EPP-AF® preserved the lung parenchyma, resulting in an increased lifespan of mice with sepsis. It can be a helpful adjuvant in prophylactic treatment with antibiotics in presurgical conditions.

Prophylactic Treatment With Simvastatin Modulates the Immune Response and Increases Animal Survival Following Lethal Sepsis Infection.

Chronic use of statins may have anti-inflammatory action, promoting immunomodulation and survival in patients with sepsis. This study aimed to analyze the effects of pretreatment with simvastatin in lethal sepsis induced by cecal ligation and puncture (CLP). Male Swiss mice received prophylactic treatment with simvastatin or pyrogen-free water orally in a single daily dose for 30 days. After this period, the CLP was performed. Naïve and Sham groups were performed as non-infected controls. Animal survival was monitored for 60 h after the CLP. Half of mice were euthanized after 12 h to analyze colony-forming units (CFUs); hematological parameters; production of IL-10, IL-12, IL-6, TNF-α, IFN-γ, and MCP-1; cell counts on peritoneum, bronchoalveolar lavage (BAL), bone marrow, spleen, and mesenteric lymph node; immunephenotyping of T cells and antigen presenting cells and production of hydrogen peroxide (H2O2). Simvastatin induced an increase in survival and a decrease in the CFU count on peritoneum and on BAL cells number, especially lymphocytes. There was an increase in the platelets and lymphocytes number in the Simvastatin group when compared to the CLP group. Simvastatin induced a greater activation and proliferation of CD4+ T cells, as well as an increase in IL-6 and MCP-1 production, in chemotaxis to the peritoneum and in H2O2 secretion at this site. These data suggest that simvastatin has an impact on the survival of animals, as well as immunomodulatory effects in sepsis induced by CLP in mice.

Therapeutic potential of biodegradable microparticles containing Punica granatum L. (pomegranate) in murine model of asthma.

OBJECTIVE AND DESIGN: Among the options for treatment of diseases affecting the respiratory system, especially asthma, drug delivering systems for intranasal application represent an important therapeutic approach at the site of inflammation. The present study aimed to evaluate the therapeutic effect of biodegradable microparticles formed by poly lactic-co-glycolic acid (PLGA) containing encapsulated pomegranate extract on a murine model of asthma. MATERIAL: The extract was acquired from the leaves of P. granatum and characterized qualitatively by HPLC. A w/o/w emulsion solvent extraction-evaporation method was chosen to prepare the microparticles containing pomegranate encapsulated extract (MP). TREATMENT: OVA-sensitized BALB/c mice were used as asthma model and treated with dexamethasone and P. granatum extract in solution form or encapsulated into microparticles. RESULTS: MP were able to inhibit leukocytes' recruitment to bronchoalveolar fluid, especially, eosinophils, decreasing cytokines (IL-1ß and IL-5) and protein levels in the lungs. CONCLUSIONS: This approach can be used as an alternative/supplementary therapy based on the biological effects of P. granatum for managing inflammatory processes, especially those with pulmonary complications.

Immune cells recruitment and activation by Tityus serrulatus scorpion venom.

Despite several studies showed that the Tityus serrulatus scorpion venom (Tsv) induces an inflammatory response, just a few have investigated the effect of the venom on the immune response. Therefore, the aim of this study was to evaluate alterations of venom application on lymphoid organs and on the recruitment and activation of cells and also on the cytokine production. Swiss male mice (2-3 months, 20-25 g) received a non-lethal dose of crude Tsv (200 µg/kg), diluted in sterile PBS by subcutaneous route. Control animals received only sterile PBS. The animals were sacrificed after 30, 120 and 360 min. The inflammatory parameters studied were skin histology at the site of venom application, leukocyte count, and blood cytokine levels (IL-6, IL-10, and TNF-α). Inguinal lymph node, spleen and bone marrow cellularity was determined for evaluation of the Tsv effect on immune system organs. The results showed that Tsv caused no local inflammation, but it induced an increase of blood neutrophils and serum IL-6, TNF-α and IL-10. After 360 min of envenomation there was a reduction in the cells number from peritoneum and spleen, but there was an increase in the cell number from lymph nodes. In conclusion, the Tsv induces systemic alterations characterized by changes in the cell number in lymphoid organs, increase pro and anti-inflammatory cytokines.

Evaluation of the subchronic toxicity of oral treatment with Chenopodium ambrosioides in mice.

AIM OF THE STUDY: The leaves of Chenopodium ambrosioides L. (Chenopodiaceae) have been used by native people to treat many diseases. Recently, we showed that the treatment with small dose (5mg/kg) of hydroalcoholic extract (HE) from Chenopodium ambrosioides' leaves has immunestimulatory effects. The aim of this study was to investigate the subchronic toxicity of the oral treatment with this HE in preclinical assays. MATERIAL AND METHODS: Swiss mice were divided into 4 groups (n=10/group). They received the HE daily at the doses of 5, 50 and 500 mg/kg by gavage during 15 days. The control group received only water. They were observed each hour for 24h and each day for 15 days, when the blood was collected. The serum was used to perform the biochemical analysis. The mice were then killed and the vital and lymphoid organs were collected and evaluated. RESULTS: There was neither death nor alterations in the body weight in the HE-treated groups, but there were alterations in the weight of some organs. There was an increase in the lymph node cells number in the highest two doses. The number of cells in the bone marrow was high in the HE-treated groups, but the number of peritoneal cells was smaller in the HE-treated groups when compared to the control. There was no alteration in the AST, but there was a reduction in the albumin levels in the HE500 group and in the triglycerides and VLDL in the highest doses. CONCLUSION: The subchronic treatment with HE induced punctual alterations in the groups treated with the highest doses. However, the HE treatment was not lethal and did not induce toxic alterations using the therapeutic dose, suggesting that it is safe to use this product in the adequate dose.