Preeclampsia (PE) is a specific syndrome of pregnancy, characterized by hypertension and proteinuria. This pathology is associated with hyperuricemia and elevated serum levels of inflammatory cytokines. Uric acid crystals may activate an intracellular complex called inflammasome, which is important for processing and release of inflammatory cytokines. This study investigated the state of monocyte activation, both endogenous and stimulated with monosodium urate (MSU), by gene expression of NLRP1 and NLRP3 receptors as well as their association with inflammatory cytokines expression. Monocytes were obtained from peripheral blood of 23 preeclamptic pregnant women, 23 normotensive pregnant women (NT) and 23 healthy non-pregnant women (NP). Inflammasome activation was evaluated by the gene expression of NLRP1, NLRP3, caspase-1, IL-1β, IL-18 and TNF-α by RT-qPCR in unstimulated monocytes (endogenous expression), or after cell stimulation with MSU (stimulated expression). The concentration of cytokines was assessed by ELISA. In preeclamptic pregnant women, gene expression of NLRP1, NLRP3, caspase-1, IL-1β and TNF-α by monocytes stimulated or not with MSU was significantly higher than in NT and NP groups. Stimulation of monocytes from preeclamptic and non-pregnant women with MSU induced increased gene expression of NLRP3, caspase-1 and TNF-α in relation to the endogenous expression in these groups, while this was not observed in the NT group. The cytokine determination showed that monocytes from women with PE produced higher endogenous levels of IL-1β, IL-18 and TNF-α compared to the other groups, while the stimulus with MSU led to higher production of these cytokines in preeclamptic group than in the NT group. In conclusion, the results showed increased basal gene expression of NLRP1 and NLRP3 receptors in monocytes from PE group. These cells stimulation with MSU demonstrates that uric acid plays a role in NLRP3 inflammasome activation, suggesting the participation of this inflammatory complex in the pathogenesis of preeclampsia.

Pre-eclampsia (PE) is an obstetric pathology characterized by abnormal activation of the innate and adaptive immune systems dependent on the imbalance of T helper subsets. The present study aimed to evaluate the gene and protein expression of T helper type 1 (Th1)/Th2/Th17/regulatory T (Treg) cell transcription factors in peripheral blood lymphocytes from pregnant women with PE employing quantitative RT-PCR and flow cytometry techniques, as well as the cytokine profile produced by these CD4 T-cell subsets in the plasma of pregnant women with PE, classified as early-onset PE (n = 20), late-onset PE (n = 20) and normotensive pregnant women (n = 20). Results showed a higher percentage of CD4 T cells expressing the RORc transcription factor (Th17) and a lower percentage of cells expressing FoxP3 (Treg) in women with early-onset PE compared with late-onset PE and normotensive groups. A lower gene expression of GATA-3 transcription factor was detected in cells of women with early-onset PE compared with the late-onset PE group. Endogenous plasma levels of interleukin-6 (IL-6), IL-17 and tumour necrosis factor-α were significantly higher in the early-onset PE group than in the late-onset PE and normotensive groups, whereas IL-4 (Th2 profile) and IL-22 (Th17 profile), were not significantly different between the studied groups. The endogenous levels of transforming growth factor-β and IL-10 were significantly lower in the pre-eclamptic than in the normotensive groups of the same gestational age, with a significant difference between early- and late-onset PE. The results show that in women with PE there is an imbalance between inflammatory and anti-inflammatory profiles in CD4 T-cell subsets, with polarization to Th17 profiles in the early-onset PE, considered as the severe form of PE.

Preeclampsia (PE) is a human pregnancy-specific syndrome with abnormal activation of cells from the innate immune system. The present study evaluated whether silibinin (SB) treatment of monocytes from preeclamptic women could modulate NLRP1 and NLRP3 inflammasomes as well as TLR4/NF-κB pathway activation. Peripheral blood monocytes from 20 preeclamptic and 20 normotensive (NT) pregnant women, as well as the THP-1 cell line, were cultured with or without monosodium urate (MSU) or SB. NLRP1, NLRP3, Caspase-1, TLR4, MyD88, NF-κB, IL-1β, IL-18, TNF-α and IL-10 gene expression by monocytes was analysed by quantitative real-time polymerase chain reaction (qPCR), while inflammatory cytokine production and p65NF-κB activity were determined by enzyme-linked immunosorbent assays (ELISAs). TLR4/MyD88/NF-κB and NLRP1/NLRP3 inflammasomes pathways in THP-1 cells were evaluated by flow cytometry and western blot respectively. Compared with NT women, monocytes from preeclamptic women showed The Ethics Committee of the Botucatu Medical School approved the study (protocol number 2.333.216)higher endogenous activation of NLRP1/NLRP3 inflammasomes and the TLR4/NF-κB pathway as well as higher gene and protein expression of IL-1β, IL-18 and TNF-α, and lower expression of IL-10. Monocyte stimulation with MSU increased inflammation-related genes as well as NF-κB activity. In vitro, SB treatment of monocytes from preeclamptic women reduced the basal activation of these cells by decreasing NLRP1/NLRP3 inflammasomes and p65NF-κB activity. THP-1 cells exhibited a similar immunological response profile to monocytes from preeclamptic women when cultured with or without MSU or SB. These results suggest uric acid participates in the systemic inflammatory response characteristic of preeclampsia and that in vitro SB treatment can modulate the sterile inflammation established in monocytes from preeclamptic women.

ResumoIntrodução: A dor no período neonatal é motivo de inúmeros estudos, sendo fundamental para a sua abordagem uma avaliação criteriosa e adequada.Objetivo: Comparar as variáveis fisiológicas com as comportamentais para a avaliação da dor em recém-nascido (RN) prematuro. Métodos: Estudo realizado entre fevereiro de 2003 e maio de 2004, com recém-nascidos de idade gestacional (IG) abaixo de 34 semanas e peso de nascimento menor que 1500 g, submetidos à ventilação mecânica e não sedados. As variáveis fisiológicas estudadas foram as freqüências cardíaca (FC) e respiratória (FR) e a saturação de oxigênio (SatO 2 ); as variáveis comportamentais foram avaliadas por meio da escala de dor do recém-nascido -Neonatal Infant Pain Scale (NIPS). Todas as variáveis foram controladas no terceiro dia de vida do RN, antes, imediatamente após e cinco minutos após o procedimento de aspiração endotraqueal. Resultados: Foram estudados 50 RNs com IG média ao nascimento de 29,98 ± 2,24 semanas e peso médio de nascimento de 1087,20 ± 350,06 g, sendo 28 (56%) RN do sexo feminino e a doença das membranas hialinas foi diagnosticada em todos os RNs estudados. Pelo teste t-Student verificou-se diferença significante na SatO 2 nos momentos estudados, sendo o mesmo não observado em relação a FC e FR. A mediana da NIPS foi maior nos momentos pós aspiração. As variáveis fisiológicas mostraram-se pouco sensíveis à detecção da dor (FC: 40,7%, FR: 24,1%, SatO 2 : 6,6%), e a escala NIPS mostrou-se mais específica para a mesma avaliação (86,6%). Conclusão: As variáveis fisiológicas apresentaram pouca sensibilidade e especificidade para a avaliação da dor no RN prematuro, quando avaliadas isoladamente. Palavras-chave:Recém-nascido; dor; avaliação da dor. AbstractIntroduction: Pain in the neonatal period is the objective of several studies, being important for its approach an adequate evaluation. Objective: To compare the physiological and behavioral variables with the behavior for assessment of pain in premature newborns (NB). Methods: Prospective study carried out from February/2003 to May/2004, including premature infants with gestational age (GA) below 34 weeks and birth weight less than 1.500 g, submitted to mechanical ventilation and not sedation. The physiological variables studied were: heart rate (HR), respiratory rate (RR) and oxygen saturation (SatO 2 ); the behavioral variables were evaluated by NIPS. All the variables have been controlled in the third day of life of NB, immediately after and five minutes after the procedure of endotracheal suction. Results: Fifty premature infants were studied with GA at birth of 29.98 ± 2.24 weeks and birth weight 1087.20 ± 350.06 g and the respiratory distress syndrome was diagnosed in all NB studied. Using t-Student test, it was verified significant difference on SatO 2 at the different moments studied; however, there was no statistical significant difference in relation to HR and RR. The median of NIPS was high on moments following suction. The physiological variables showed: low sensitivity on detec...

Pre-eclampsia (PE) is a human pregnancy syndrome with abnormal activation of the innate immune response. The study evaluated the involvement of molecular structures called damage-associated molecular patterns (DAMPs), such as hyaluronan (HA) and heat shock proteins (Hsp) on NLRP1 and NLRP3 inflammasomes activation in peripheral blood monocytes. Twenty pre-eclamptic women, 20 normotensive pregnant women (NT) and 20 non-pregnant women (NP) were studied. Enzyme immunoassay was employed for the determination of HA, Hsp70 and High mobility group Box 1 (HMGB1) in plasma, as well as for the detection of Interleukin-1β (IL-1β), IL-18 and tumor necrosis factor alpha (TNF-α) in the supernatant of monocytes cultured with or without HA and Hsp70. The inflammasomes induction was evaluated by the quantification of mRNA for NLRP1, NLRP3, caspase-1, IL-1β, IL-18, HMGB1 and TNF-α by qPCR in monocyte culture. The results showed significantly higher plasma levels of HA, Hsp70 and HMGB1 in pre-eclamptic women than in NT and NP women. Monocytes from women with PE showed endogenous activation of NLRP1 and NLRP3 inflammasomes, and expressed high amounts of IL-1β, IL-18, HMGB1 and TNF-α. The stimulation of monocytes with HA increased the gene expression of NLRP1, NLRP3, caspase-1, TNF-α, IL-1β, HMGB1 and IL-18 and the production of IL-1β in pre-eclamptic women. Monocytes cultured with Hsp70 produced elevated levels of IL-1β and TNF-α through a mechanism independent of inflammasomes activation. These results suggest the participation of these DAMPs in the systemic inflammatory response that is characteristic of PE.