Mygalin, an Acanthoscurria gomesiana spider-derived synthetic, modulates haloperidol-induced cataleptic state in mice.

BACKGROUND: Haloperidol (HAL) is an antipsychotic used in the treatment of schizophrenia. However, adverse effects are observed in the extrapyramidal tracts due to its systemic action. Natural compounds are among the treatment alternatives widely available in Brazilian biodiversity. Mygalin (MY), a polyamine that was synthesized from a natural molecule present in the hemolymph of the Acanthoscurria gomesian spider, may present an interesting approach. AIMS: This study aimed to evaluate the effect of MY in mice subjected to HAL-induced catalepsy. METHODS: Male Swiss mice were used. Catalepsy was induced by intraperitoneal administration of HAL (0.5 mg/kg - 1 mL/Kg) diluted in physiological saline. To assess the MY effects on catalepsy, mice were assigned to 4 groups: (1) physiological saline (NaCl 0.9 %); (2) MY at 0.002 mg/Kg; (3) MY at 0.02 mg/Kg; (4) MY at 0.2 mg/Kg. MY or saline was administered intraperitoneally (IP) 10 min b HAL before saline. Catalepsy was evaluated using the bar test at 15, 30, 60, 90, and 120 min after the IP administration of HAL. RESULTS: The latency time in the bar test 15, 30, 60, and 90 min increased (p < 0.05) after IP administration of HAL compared to the control group. Catalepsy was attenuated 15, 30, 90, and 120 min (p < 0.05) after the IP-administration of MY at 0.2 mg/Kg; while MY at 0.02 mg/Kg attenuated catalepsy 15 min after the HAL treatment. Our findings showed that MY attenuates the HAL-induced cataleptic state in mice.

Mygalin, an Acanthoscurria gomesiana spider‐derived synthetic, modulates haloperidol-induced cataleptic state in mice

Background: Haloperidol (HAL) is an antipsychotic used in the treatment of schizophrenia. However, adverse effects are observed in the extrapyramidal tracts due to its systemic action. Natural compounds are among the treatment alternatives widely available in Brazilian biodiversity. Mygalin (MY), a polyamine that was synthesized from a natural molecule present in the hemolymph of the Acanthoscurria gomesian spider, may present an interesting approach. Aims: This study aimed to evaluate the effect of MY in mice subjected to HAL-induced catalepsy. Methods: Male Swiss mice were used. Catalepsy was induced by intraperitoneal administration of HAL (0.5 mg/kg − 1 mL/Kg) diluted in physiological saline. To assess the MY effects on catalepsy, mice were assigned to 4 groups: (1) physiological saline (NaCl 0.9 %); (2) MY at 0.002 mg/Kg; (3) MY at 0.02 mg/Kg; (4) MY at 0.2 mg/Kg. MY or saline was administered intraperitoneally (IP) 10 min b HAL before saline. Catalepsy was evaluated using the bar test at 15, 30, 60, 90, and 120 min after the IP administration of HAL. Results: The latency time in the bar test 15, 30, 60, and 90 min increased (p < 0.05) after IP administration of HAL compared to the control group. Catalepsy was attenuated 15, 30, 90, and 120 min (p < 0.05) after the IP-administration of MY at 0.2 mg/Kg; while MY at 0.02 mg/Kg attenuated catalepsy 15 min after the HAL treatment. Our findings showed that MY attenuates the HAL-induced cataleptic state in mice.

In Silico and In Vitro Approach for Evaluation of the Anti-Inflammatory and Antioxidant Potential of Mygalin.

There is a great interest in describing new molecules to be used as therapeutic targets in various diseases, particularly those that play a role in inflammatory responses and infection control. Mygalin is a synthetic analogue of spermidine, and previous studies have demonstrated its bactericidal effect against Escherichia coli, as well as its ability to modulate the inflammatory response of macrophages against lipopolysaccharide (LPS). However, the mechanisms through which mygalin regulates this inflammatory response remain poorly characterized. A set of platforms using molecular docking analysis was employed to analyze various properties of mygalin, including toxicity, biodistribution, absorption, and the prediction of its anti-inflammatory properties. In in vitro assays, we evaluated the potential of mygalin to interact with products of the inflammatory response, such as reactive oxygen species (ROS) and antioxidant activity, using the BMDM cell. The in silico analyses indicated that mygalin is not toxic, and can interact with proteins from the kinase group, and enzymes and receptors in eukaryotic cells. Molecular docking analysis showed interactions with key amino acid residues of COX-2, iNOS and 5-LOX enzymes. In vitro, assays demonstrated a significant reduction in the expression of iNOS and COX-2 induced by LPS, along with a decrease in the oxidative stress caused by the treatment with PMA, all without altering cell viability. Mygalin exhibited robust antioxidant activity in DPPH assays, regardless of the dose used, and inhibited heat-induced hemolysis. These studies suggest that mygalin holds promise for further investigation as a new molecule with anti-inflammatory and antioxidant properties.

Acanthoscurria gomesiana spider-derived Mygalin in the prelimbic prefrontal cortex modulates neuropathic pain and depression comorbid.

Depression has a high rate of comorbidity with neuropathic pain. This study aims to investigate the effect of Mygalin, an acylpolyamine synthesized from a natural molecule in the hemolymph of the Acanthoscurria gomesiana spider, injected into the prelimbic (PrL) region of the medial prefrontal cortex on chronic neuropathic pain and depression comorbidity in rats. To investigate that comorbidity, neuropathic pain was induced by chronic constriction injury (CCI) of the sciatic nerve in male Wistar rats. The biotinylated biodextran amine (BDA) bidirectional neural tract tracer was microinjected into the PrL cortex to study brain connections. Rodents were further subjected to von Frey (mechanical allodynia), acetone (cold allodynia), and forced swim (depressive-like behavior) tests. BDA neural tract tracer-labeled perikarya were found in the dorsal columns of the periaqueductal gray matter (dPAG) and the dorsal raphe nucleus (DRN). Neuronal activity of DRN neurons decreased in CCI rats. However, PrL cortex treatment with Mygalin increased the number of spikes on DRN neurons. Mygalin treatment in the PrL cortex decreased both mechanical and cold allodynia and immobility behavior in CCI rats. PrL cortex treatment with N-methyl-D-aspartate (NMDA) receptor receptors attenuated the analgesic and antidepressive effects caused by Mygalin. The PrL cortex is connected with the dPAG and DRN, and Mygalin administration into the PrL increased the activity of DRN neurons. Mygalin in the PrL cortex produced antinociceptive and antidepressive-like effects, and the NMDA agonist reversed these effects.

In silico and In Vitro approach for evaluation of the anti-inflammatory and antioxidant potential of mygalin

There is a great interest in describing new molecules to be used as therapeutic targets in various diseases, particularly those that play a role in inflammatory responses and infection control. Mygalin is a synthetic analogue of spermidine, and previous studies have demonstrated its bactericidal effect against Escherichia coli, as well as its ability to modulate the inflammatory response of macrophages against lipopolysaccharide (LPS). However, the mechanisms through which mygalin regulates this inflammatory response remain poorly characterized. A set of platforms using molecular docking analysis was employed to analyze various properties of mygalin, including toxicity, biodistribution, absorption, and the prediction of its anti-inflammatory properties. In in vitro assays, we evaluated the potential of mygalin to interact with products of the inflammatory response, such as reactive oxygen species (ROS) and antioxidant activity, using the BMDM cell. The in silico analyses indicated that mygalin is not toxic, and can interact with proteins from the kinase group, and enzymes and receptors in eukaryotic cells. Molecular docking analysis showed interactions with key amino acid residues of COX-2, iNOS and 5-LOX enzymes. In vitro, assays demonstrated a significant reduction in the expression of iNOS and COX-2 induced by LPS, along with a decrease in the oxidative stress caused by the treatment with PMA, all without altering cell viability. Mygalin exhibited robust antioxidant activity in DPPH assays, regardless of the dose used, and inhibited heat-induced hemolysis. These studies suggest that mygalin holds promise for further investigation as a new molecule with anti-inflammatory and antioxidant properties.

Efeito da migalina sobre a resposta inflamatória provocada por monofosforil lipídio A (MFLA), agonista de TLR4, na linhagem Raw 264.7

Macrophages are important cells of the immune system that play a role in innate and adaptive immune responses. They can differentiate into M1 and M2 subpopulations that display distinct functions in response to different stimuli. Toll-like receptors are fundamental in recognizing molecules associated with pathogens, as they activate macrophages and the innate immune response. The use of toll-like receptor activating molecules has been studied as a potential for new adjuvants, among these molecules Monophosphoryl lipid A (MPLA) stands out, a detoxified derivative of Lipid A, the main component of LPS (TLR4 agonist). Mygalin is a synthetic acylpolamine, analogous to the spermidine polyamine, originating from hemocytes of the spider Acanthoscurria gomesiana. This molecule has proven microbicide activity and does not cause cytotoxicity in eukaryotic cells. Mygalin binds to E. coli LPS and regulates the overproduction of pro-inflammatory cytokines in response to LPS. MPLA is one of the detoxified LPS subunits and has been used as an adjuvant. We investigated, using in vitro assays, the immunomodulatory character of Mygalin on the inflammatory response induced by the activation of macrophages of the Raw 264.7 cell line in response to MPLA, to define the contribution of this subunit in the suppression of the response to LPS. LPS and IFN-γ were used in parallel and possible mechanisms of this immune regulation were explored. We investigated, using in vitro assays, the immunomodulatory character of Mygalin on the inflammatory response induced by the activation of macrophages of the Raw 264.7 cell line in response to MPLA, to define the contribution of this subunit in the suppression of the response to LPS. LPS and IFN-γ were used in parallel and possible mechanisms of this immune regulation were explored. Our results showed that Mygalin did not induce NO synthesis in macrophages stimulated with both MPLA and LPS and suppressed the synthesis of cytokine IL-6 and chemokine CCL2 in cells stimulated with these TLR4 agonists. The contribution of the arginase activation pathway in NO suppression was also evaluated using NOHA as an inhibitor, the direct action of this enzyme in reducing NO synthesis was not proven.

Os macrófagos são importantes células do sistema imunológico que atuam na resposta imune inata e adaptativa. São capazes de se diferenciar em subpopulações M1 e M2 que apresentam funções distintas em resposta a diferentes estímulos. Os receptores Toll-like são fundamentais no reconhecimento das moléculas associadas aos patógenos, pois ativam os macrófagos e a resposta imune inata. O uso de moléculas ativadoras de receptor toll-like tem sido estudado como potencial de novos adjuvantes, dentre essas moléculas destaca-se o Monofosforil lipídeo A (MFLA), um derivado desintoxicado do Lipídio A, o principal componente do LPS (agonista de TLR4). A Migalina é uma acilpoliamina sintética, análoga da poliamina espermidina, originária de hemócitos da aranha Acanthoscurria gomesiana. Esta molécula possui comprovada atividade microbicida e não provoca citotoxicidade nas células eucarióticas. A Migalina se liga ao LPS de E. coli e regula o excesso de produção de citocinas pro-inflamatória em resposta ao LPS. O MFLA é uma das subunidades detoxificada do LPS, e vem sendo usado como adjuvante. Neste trabalho investigamos em ensaios in vitro o caráter imunomodulador da Migalina sobre a resposta inflamatória induzida pela ativação de macrófagos de linhagem celular Raw 264.7 em resposta ao MFLA, para definir a contribuição desta subunidade na supressão da resposta ao LPS. Foi usado paralelamente LPS e IFN-γ e explorado possíveis mecanismos desta regulação imune. Nossos resultados mostraram que a Migalina não induziu a síntese de NO em macrófagos estimulados tanto com MFLA quanto LPS e suprimiu a síntese da citocina IL-6 e da quimiocina CCL2 em células estimuladas com estes agonistas de TLR4. Foi também avaliado a contribuição da via de ativação da arginase na supressão de NO usando o inibidor NOHA, não sendo comprovada a ação direta desta enzima na redução da síntese de NO.

Acanthoscurria gomesiana spider-derived synthetic mygalin in the dorsal raphe nucleus modulates acute and chronic pain.

Mygalin, a diacylspermidine that is naturally found in the hemolymph of the spider Acanthoscurria gomesiana, is of interest for development as a potential analgesic. Previous studies have shown that acylpolyamines modulate glutamatergic receptors with the potential to alter pain pathways. This study aimed to evaluate the effects of mygalin on acute and chronic pain in rodents. For evaluation of acute pain, Wistar rats were subjected to tail-flick and hot-plate nociceptive tests. For the evaluation of chronic neuropathic pain, a partial ligation of the sciatic nerve was performed and, 21 days later, animals were examined in hot-plate, tail-flick, acetone, and von Frey tests. Either Mygalin or vehicle was microinjected in the dorsal raphe nucleus (DRN) before the tests. Another group was pretreated with selective antagonists of glutamate receptors (LY 235959, MK-801, CNQX, and NBQX). Mygalin decreases nociceptive thresholds on both acute and chronic neuropathic pain models in all the tests performed. The lowest dose of mygalin yielded the most effective nociception, showing an increase of 63% of the nociceptive threshold of animals with neuropathic chronic pain. In conclusion, mygalin microinjection in the DRN results in antinociceptive effect in models of neuropathic pain, suggesting that acylpolyamines and their derivatives, such as this diacylspermidine, could be pursued for the treatment of neuropathic pain and development of selective analgesics.

Acylpolyamine Mygalin as a TLR4 Antagonist Based on Molecular Docking and In Vitro Analyses.

Toll-like receptors (TLRs) are transmembrane proteins that are key regulators of innate and adaptive immune responses, particularly TLR4, and they have been identified as potential drug targets for the treatment of disease. Several low-molecular-weight compounds are being considered as new drug targets for various applications, including as immune modulators. Mygalin, a 417 Da synthetic bis-acylpolyamine, is an analog of spermidine that has microbicidal activity. In this study, we investigated the effect of mygalin on the innate immune response based on a virtual screening (VS) and molecular docking analysis. Bone marrow-derived macrophages and the cell lines J774A.1 and RAW 264.7 stimulated with lipopolysaccharide (LPS) were used to confirm the data obtained in silico. Virtual screening and molecular docking suggested that mygalin binds to TLR4 via the protein myeloid differentiation factor 2 (MD-2) and LPS. Macrophages stimulated by mygalin plus LPS showed suppressed gene expression of tumor necrosis factor (TNF-α), interleukine 6 (IL-6), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), as well as inhibition of signaling protein p65 of the nuclear factor κB (NF-κB), resulting in decreased production of nitric oxide (NO) and TNF-α. These results indicate that mygalin has anti-inflammatory potential, being an attractive option to be explored. In addition, we reinforce the importance of virtual screening analysis to assist in the discovery of new drugs.

Mygalin: an acylpolyamine with bactericidal activity

Inappropriate use of antibiotics favors the selection and spread of resistant bacteria. To reduce the spread of these bacteria, finding new molecules with activity is urgent and necessary. Several polyamine analogs have been constructed and used to control microorganisms and tumor cells. Mygalin is a synthetic acylpolyamine, which are analogs of spermidine, derived from the hemolymph of the spider Acanthoscurria gomesiana. The effective activity of polyamines and their analogs has been associated with their structure. The presence of two acyl groups in the Mygalin structure may give this molecule a specific antibacterial activity. The aim of this study was to identify the mechanisms involved in the interaction of Mygalin with Escherichia coli to clarify its antimicrobial action. The results indicated that Mygalin exhibits intense dose and time-dependent bactericidal activity. Treatment of E. coli with this molecule caused membrane rupture, inhibition of DNA synthesis, DNA damage, and morphological changes. The esterase activity increased along with the intracellular production of reactive oxygen species (ROS) after treatment of the bacteria with Mygalin. In addition, this molecule was able to sequester iron and bind to LPS. We have shown that Mygalin has bactericidal activity with underlying mechanisms involving ROS generation and chelation of iron ions that are necessary for bacterial metabolism, which may contribute to its microbicidal activity. Taken together, our data suggest that Mygalin can be explored as a new alternative drug with antimicrobial potential against Gram-negative bacteria or other infectious agents.

Acylpolyamine mygalin as a TLR4 antagonist based on molecular docking and In vitro analyses

Toll-like receptors (TLRs) are transmembrane proteins that are key regulators of innate and adaptive immune responses, particularly TLR4, and they have been identified as potential drug targets for the treatment of disease. Several low-molecular-weight compounds are being considered as new drug targets for various applications, including as immune modulators. Mygalin, a 417 Da synthetic bis-acylpolyamine, is an analog of spermidine that has microbicidal activity. In this study, we investigated the effect of mygalin on the innate immune response based on a virtual screening (VS) and molecular docking analysis. Bone marrow-derived macrophages and the cell lines J774A.1 and RAW 264.7 stimulated with lipopolysaccharide (LPS) were used to confirm the data obtained in silico. Virtual screening and molecular docking suggested that mygalin binds to TLR4 via the protein myeloid differentiation factor 2 (MD-2) and LPS. Macrophages stimulated by mygalin plus LPS showed suppressed gene expression of tumor necrosis factor (TNF-α), interleukine 6 (IL-6), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), as well as inhibition of signaling protein p65 of the nuclear factor κB (NF-κB), resulting in decreased production of nitric oxide (NO) and TNF-α. These results indicate that mygalin has anti-inflammatory potential, being an attractive option to be explored. In addition, we reinforce the importance of virtual screening analysis to assist in the discovery of new drugs.

Antibacterial and antitumoral activities of the spider acylpolyamine mygalin silver nanoparticles

Mygalin is a synthetic analog of polyamine spermidine isolated from spider hemocytes. Polyamines show potential therapeutic activity against a wide range of human diseases such as cancer and microbial infections. In this work, we analyzed the antibacterial and antitumoral activities of Mygalin silver nanoparticles synthesized by the photoreduction method. The formation and distribution of MygAgNPs were confirmed by UV-visible spectroscopy, zeta potential, and transmission electron microscopy. The obtained nanoparticles were mostly spherical with a particle size distribution in the range of ~ 10–60 nm. We have demonstrated that MygAgNPs increased the effectiveness of the native Mygalin by approximately 6400-fold. Cytotoxicity tests were performed, and it was possible to reach a concentration that was not toxic to healthy cells (NHI-3T3) and at the same time toxic to the tumor cell line (MCF-7). The obtained results suggest that this system shows potential enhanced antibacterial activity against Escherichia coli, DH5a and anticancer activity against MCF-7 cell line

Mygalin: An Acylpolyamine With Bactericidal Activity.

Inappropriate use of antibiotics favors the selection and spread of resistant bacteria. To reduce the spread of these bacteria, finding new molecules with activity is urgent and necessary. Several polyamine analogs have been constructed and used to control microorganisms and tumor cells. Mygalin is a synthetic acylpolyamine, which are analogs of spermidine, derived from the hemolymph of the spider Acanthoscurria gomesiana. The effective activity of polyamines and their analogs has been associated with their structure. The presence of two acyl groups in the Mygalin structure may give this molecule a specific antibacterial activity. The aim of this study was to identify the mechanisms involved in the interaction of Mygalin with Escherichia coli to clarify its antimicrobial action. The results indicated that Mygalin exhibits intense dose and time-dependent bactericidal activity. Treatment of E. coli with this molecule caused membrane rupture, inhibition of DNA synthesis, DNA damage, and morphological changes. The esterase activity increased along with the intracellular production of reactive oxygen species (ROS) after treatment of the bacteria with Mygalin. In addition, this molecule was able to sequester iron and bind to LPS. We have shown that Mygalin has bactericidal activity with underlying mechanisms involving ROS generation and chelation of iron ions that are necessary for bacterial metabolism, which may contribute to its microbicidal activity. Taken together, our data suggest that Mygalin can be explored as a new alternative drug with antimicrobial potential against Gram-negative bacteria or other infectious agents.

Efeito da Migalina sobre atividade inflamatória com diferentes estímulos em linhagens de macrófagos J774 e derivados de medula óssea

Natural polyamines putrescine, spermine and spermidine are essential for cell growth and proliferation, protein and nucleic acid synthesis, cell matrix repair, adhesion and signaling processes. Mygalin was isolated from hemocytes of the spider Acanthoscurria gomesiana and identified as a bis-acylpolyamine analogue to spermidine, possessing microbicidal activity. Macrophages are innate immune response cells fundamental in the inflammatory process and are programmed to detect molecular patterns associated with pathogens. Immune cell surface molecules and cytokines are the key components for the achievement of the inflammatory response. Due to the limited knowledge of the role of Mygalin in the immune response, we investigated its immunomodulatory function in macrophages J774 and macrophages derived from C57BL / 6 mouse marrow MODM against several stimuli in different assays. The cells were either preincubated with Mygalin 40-160 μg/ml for 2 hours prior to stimulation or not with LPS 100 ng/ml, Toxoplasma gondii antigen 10-25-50 μg/ml, IFN-γ 1 ng/ml and Zimosan 10 μg/ml After 24 hours, hydrogen peroxide production was examined by the fluorimetric method with and without PMA (200 nM) and the alteration of the expression of the surface molecules by flow cytometry FACS. Supernatants from the J774 and MODM assays were harvested for quantify production of IL-12p40 ELISAand nitrite Griess. Treatment of cells with Mygalin and Toxoplasma gondii didn’t induce IL-12p40 and NO production. In J774 IFN-γ didn’t change the amount of NO in the control, but the LPS increased even in the presence of Mygalin and the same happened with LPS+IFN-γ. Zimosan increased NO production in J774, but IFN- γ reduced it. In MODM independent of the stimulus there was no alteration of the NO synthesis compared to control. Mygalin didn’t increase the production of IL-12p40 when the cells were stimulated by LPS and LPS+IFN-γ. Zimosan induced the synthesis of IL-12p40, but with IFN-γ the production reduced. We evaluated the effect of Mygalin on the production of hydrogen peroxide in MODM that Migalina reduced H2O2 production in the presence of PMA. Regarding the expression of CD11b: in J774 the LPS and LPS+IFN-γ decreased expression; F4 / 80: in MODM the LPS increased its synthesis; CD86: in J774 the LPS decreased expression even with IFN-γ, and in MODMall stimuli reduced synthesis even with Mygalin; CD284: no change in J774cells; CD206: LPS and LPS+IFN-γ there was a slight increase of the synthesis.The mechanisms involved are being analyzed for a better characterization ofthe role of macrolide in the macrophage.

Poliaminas naturais (putrescina, espermina e espermidina) são fundamentais para o crescimento e proliferação celular, na síntese de proteínas e de ácidos nucleicos, reparação da matriz celular, adesão e processos de sinalização. A Migalina foi isolada de hemócitos da aranha Acanthoscurria gomesiana e identificada como uma bis-acilpoliamina análoga a espermidina, possuindo atividade microbicida. Macrófagos são células da resposta imune inata fundamentais no processo inflamatório e são programadas para detectar padrões moleculares associados a patógenos. Moléculas de superfície de células imunes e as citocinas são componentes fundamentais para a realização da resposta inflamatória. Devido ao limitado conhecimento do papel da Migalina na resposta imune investigamos sua função imunomoduladora em macrófagos J774 e macrófagos derivados de medula de camundongo C57BL/6 (MODM) frente a vários estímulos em diferentes ensaios. As células foram pré- incubadas ou não com Migalina (40-160 μg/mL) por 2 horas antes da estimulação ou não com LPS (100 ng/mL), antígeno de Toxoplasma gondii (10- 25-50 μg/mL), IFN-γ (1 ng/mL) e Zimosan (10 μg/mL). Após 24 horas, a produção de peróxido de hidrogênio foi examinada pelo método fluorimétrico com e sem PMA (200 nM) e a alteração da expressão de moléculas de superfície por meio de citometria de fluxo (FACS). Os sobrenadantes dos ensaios de J774 e MODM foram colhidos para análises de IL-12p40 (ELISA) e nitrito (Griess). O tratamento das células com Migalina e Toxoplasma gondii não induziu a produção IL-12p40 e de NO. Em J774 o IFN-γ não alterou a quantidade de NO do controle, mas o LPS aumentou mesmo na presença de Migalina e o mesmo ocorreu com LPS+IFN-γ. O Zimosan aumentou a produção de NO em J774, mas o IFN-γ a reduziu. Em MODM independente do estímulo não houve alteração da síntese de NO comparado ao controle. A Migalina não aumentou a produção de IL-12p40 quando as células foram estimuladas com LPS e LPS+IFN-γ. Zimosan induziu a síntese de IL-12p40, mas na presença de IFN-γ a produção reduziu. Avaliamos o efeito da Migalina na produção de peróxido de hidrogênio em MODM no qual a Migalina reduziu a produção de H2O2 na presença de PMA. Com relação à expressão de CD11b: em J774 o LPS e LPS+IFN-γ diminuiu a expressão; F4/80: em MODM LPS aumentou sua síntese; CD86: em J774 LPS diminuiu a expressão mesmo com IFN-γ e em MODM todos os estímulos reduziu a síntese mesmo com Migalina; CD284: sem alteração nas células J774; CD206: LPS e LPS+IFN-γ houve um leve aumento da síntese. Os mecanismos envolvidos estão sendo analisados para melhor caracterização do papel da Migalina em macrófagos.