Biology subjects

Faccioli, L. H.

Publications and source records attributed to Faccioli, L. H..

4 recordsLinked to original sources

bioRxiv2022

Polyunsaturated fatty acids alter the formation of lipid droplets and eicosanoid production in Leishmania promatigotes

Leishmania parasites contain lipid droplets (LD, or lipid bodies) and the molecular machinery responsible for synthesizing prostaglandins (PGs) and other bioactive lipids. We studied the effects of polyunsaturated fatty acids (PUFA) on LD biogenesis and eicosanoid production in distinct Leishmania species associated with different clinical forms of leishmaniasis. We also compared structural models of human-like cyclooxygenase-2 (GP63) and prostaglandin F synthase (PGFS) proteins of Leishmania, and we evaluated their enzymatic production in logarithmic and stationary growth phases of procyclic L. amazonensis, L. braziliensis and L. infantum. PUFAs modulate the formation of LDs in L. braziliensis and L. infantum. Leishmania species with equivalent clinical manifestations and tissue tropism had same protein mutations in GP63 and PGFS. No differences in GP63 production were observed among L. amazonensis, L. braziliensis and L. infantum, however increased PGFS production was detected during the parasite differentiation. Stimulation with arachidonic acid resulted in highly elevated production of hydroxyeicosatetraenoic acids compared to prostaglandins quantified by LC-MS/MS. The present findings open new perspectives on the role of eicosanoid metabolism in Leishmania and could contribute to the development of novel antiparasitic drugs.

microbiology↗

bioRxiv2021

Cannabidiol as an add-on therapy to overcome the slow-onset and, possibly, resistance to antidepressant treatment: involvement of NAPE-PLD in the medial prefrontal cortex

Antidepressant drugs are the first-line treatment for chronic stress-related psychiatric disorders such as major depressive disorder, anxiety disorders, and post-traumatic stress disorder. However, their delayed-onset of therapeutic action, frequently occurring side effects, and incomplete clinical efficacy impose significant challenges for clinicians and patients adherence to treatment. Cannabidiol (CBD) is a major non-psychotomimetic phytocannabinoid with a wide range of potential clinical applications such as either a standalone drug or as an add-on treatment. In our study, we found that in chronically stressed male mice, CBD (30 mg/kg) rapidly induced behavioral improvement within 7 days, which was quicker than the high dose of escitalopram (ESC, 14 days). Additionally, repeated administration of a low and initially ineffective dose of CBD (7.5 mg/kg) potentiated the anti-stress effects of ESC (10 mg/kg) in mice subjected to 10 or 21 days of chronic unpredictable stress (CUS). Furthermore, our results suggested the involvement of N-acyl phosphatidylethanolamine phospholipase (NAPE-PLD) located in the prefrontal cortex (PFC) in the anti-stress effects of the 7-day treatment with ESC + CBD. This combination restored CUS-induced decreased expression of NAPE-PLD in the PFC. The behavioral effects of ESC + CBD were not observed in either constitutive NAPE-PLD knockout (KO) mice or mice with a CRISPR/Cas9-induced deletion of NAPE-PLD in the PFC. ESC + CBD treatment facilitated NAPE-PLD expression in parvalbumin (PV) interneurons in the PFC. As a conclusion, we suggest that CBD might be useful as an add-on therapy to optimize the action of (SSRI-)antidepressants, possibly by restoring the inhibitory/excitatory balance of the PFC via NAPE-PLD-mediated signaling. HighlightsO_LICBD (7.5 mg/kg) reduces the latency for anti-stress effects of escitalopram (ESC) C_LIO_LIESC + CBD increases neuroplasticity in the prefrontal cortex (PFC) C_LIO_LIESC + CBD reverses stress-induced loss of NAPE-PLD in PFC-Parvalbumin (PV)+ interneurons. C_LIO_LINAPE-PLD in the PFC participates in the anti-stress effects of ESC + CBD. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=151 HEIGHT=200 SRC="FIGDIR/small/441143v2_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@d61ef2org.highwire.dtl.DTLVardef@189e2ecorg.highwire.dtl.DTLVardef@1910213org.highwire.dtl.DTLVardef@11f6bbf_HPS_FORMAT_FIGEXP M_FIG Cannabidiol (CBD) enhances the antidepressant-like effects of escitalopram (ESC) in chronically stressed mice. While an effective dose of CBD (30mg/kg) alone rapidly improved stress-related behaviors when compared to a high dose of ESC (20mg/kg), ESC+CBD combination in sub-effective doses potentiated anti-stress responses and restored prefrontal cortex (PFC) function. These effects depended on N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD) activity within PFC parvalbumin interneurons, highlighting NAPE-PLD-mediated signaling as a key mechanism by which CBD may optimize antidepressant efficacy and reestablish inhibitory/excitatory balance in the PFC. C_FIG Chemical compounds used in this articleCannabidiol (PubChem CID: 644019); Escitalopram oxalate (PubChem CID: 146571); URB597 (PubChem CID: 1383884); Ketamine hydrochloride (PubChem CID: 15851); xylazine hydrochloride (PubChem CID: 68554); 2,2,2-Tribromoethanol (PubChem CID: 6400); Flunixin meglumine (PubChem CID: 39212); Lidocaine hydrochloride (PubChem CID: 6314); Amoxicillin (PubChem CID: 33613).

pharmacology and toxicology↗

bioRxiv2021

Drug repurposing to face Covid-19: Celastrol, a potential leading drug capable of inhibiting SARS-CoV-2 replication and induced inflammation

The global emergence of Covid-19 has caused huge human casualties. Clinical manifestations of the disease vary from asymptomatic to lethal, and the symptomatic form can be associated with cytokine storm and non-homeostatic inflammatory response. In face of the urgent demand for effective drugs to treat Covid-19, we have searched for candidate compounds using a drug repurposing approach based on in silico analysis followed by biological validation. Here we identified celastrol, a pentacyclic triterpene isolated from Tripterygium wilfordii Hook F - a plant used in traditional Chinese medicine - as one of the best compounds out of 39 repurposed drug candidates. Celastrol reverted gene expression signature from SARS-CoV-2-infected cells; bound with high-affinity energy to viral molecular targets such as main protease (Mpro) and receptor-biding domain (RBD); inhibited SARS-CoV-2 replication in monkey (Vero and Vero-ACE2) and human (Caco-2 and Calu-3) cell lines; and decreased interleukin-6 (IL-6) secretion in SARS-CoV-2-infected human cell lines. Interestingly, celastrol acted in a concentration-dependent manner, with undetectable signs of cytotoxicity. Therefore, celastrol is a promising lead drug candidate to treat Covid-19 due to its ability to suppress SARS-CoV-2 replication and IL-6 production in infected cells, two critical events in the pathophysiology of this disease.

immunology↗

bioRxiv2020

Inhibition of Osteoclast Differentiation and Activation by Leukotriene B4 Loaded in Microspheres

AO_SCPLOWBSTRACTC_SCPLOWO_ST_ABSAimC_ST_ABSLeukotriene B4 (LTB4) is a labile inflammatory lipid mediator important for host defense. We hypothesised that sustained delivery of LTB4 would be a therapeutic strategy to prevent osteoclast cell differentiation in bone resorption in inflammatory diseases. Therefore, the aim of this study was to investigate the role of LTB4 in differentiation of monocytic lineage cells into osteoclasts after stimulation with LTB4 loaded in microspheres (MS). DesignLTB4-MS were prepared using an oil-in-water emulsion solvent extraction-evaporation process. Sterility, LPS contamination, characterization and efficiency of LTB4 encapsulation were investigated. J774A.1 cells were cultured in the presence of monocyte colony stimulating factor (M-CSF) and ligand for receptor activator of nuclear factor kappa B (RANKL) and then stimulated with LTB4-MS. Cytotoxicity was determined by lactate dehydrogenase assay, osteoclast formation by means of the activity of tartrate-resistant acid phosphatase enzyme and gene expression was measured by quantitative reverse transcription polymerase chain reaction to investigate regulation of Alox5, Alox5ap, Acp5, Mmp9, Calcr and Ctsk. ResultsWe found that 5-lipoxygenase pathway is involved in the osteoclastic differentiation hematopoietic lineage cells and that exogenous addition of LTB4-MS inhibited osteoclastogenesis induced by M-CSF and RANKL. The mechanism of LTB4-MS involved induction of Mmp9 gene expression and inhibition of Calcr and Ctsk, without changing Acp5. ConclusionLTB4-MS inhibited differentiation of macrophages into an osteoclastic phenotype and cell activation under M-CSF and RANKL stimulus shedding light on a potential therapeutic strategy to prevent osteoclast differentiation.

cell biology↗