HMGB1 and inflammatory cytokines in experimental acute lung injury induced in rabbits / [HMGB1 e citocinas inflamatórias em lesão pulmonar aguda experimental induzida em coelhos]

The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.(AU)

O objetivo deste trabalho é determinar os níveis de HMGB1, TNF-alfa e IL-8 no lavado broncoalveolar (BAL), bem como quantificar a expressão sérica de TLR2 e TLR4 mRNA em tecido pulmonar de coelhos com dois graus de lesão pulmonar aguda (LPA). Os animais foram distribuídos aleatoriamente em grupos com LPA grave (S) e leve / moderada (MM), induzidas com solução salina morna, e um grupo controle. HMGB1, TNF-alfa, IL-8, TLR2mRNA e TLR4mRNA foram medidos após a indução de LPA e quatro horas de ventilação mecânica. Os resultados mostraram níveis aumentados de IL-8, TNF-alfa, HMGB1 e TLR4mRNA nos grupos com LPA. As concentrações de HMGB1, IL-8 e TNF-alfa no LBA foram maiores no S comparado ao MM. Aumento de TLR4mRNA foi observado em S e MM versus controle. Os resultados sugerem uma participação precoce da HMGB1 na LPA em conjunto com IL-8 e TNF-alfa e associação com a gravidade da LPA. O TLR4 foi expresso na ALI e possivelmente possui papel precoce na fisiopatologia da LPA, mas sem associação com a gravidade.(AU)

HMGB1 and inflammatory cytokines in experimental acute lung injury induced in rabbits / [HMGB1 e citocinas inflamatórias em lesão pulmonar aguda experimental induzida em coelhos]

The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.(AU)

O objetivo deste trabalho é determinar os níveis de HMGB1, TNF-alfa e IL-8 no lavado broncoalveolar (BAL), bem como quantificar a expressão sérica de TLR2 e TLR4 mRNA em tecido pulmonar de coelhos com dois graus de lesão pulmonar aguda (LPA). Os animais foram distribuídos aleatoriamente em grupos com LPA grave (S) e leve / moderada (MM), induzidas com solução salina morna, e um grupo controle. HMGB1, TNF-alfa, IL-8, TLR2mRNA e TLR4mRNA foram medidos após a indução de LPA e quatro horas de ventilação mecânica. Os resultados mostraram níveis aumentados de IL-8, TNF-alfa, HMGB1 e TLR4mRNA nos grupos com LPA. As concentrações de HMGB1, IL-8 e TNF-alfa no LBA foram maiores no S comparado ao MM. Aumento de TLR4mRNA foi observado em S e MM versus controle. Os resultados sugerem uma participação precoce da HMGB1 na LPA em conjunto com IL-8 e TNF-alfa e associação com a gravidade da LPA. O TLR4 foi expresso na ALI e possivelmente possui papel precoce na fisiopatologia da LPA, mas sem associação com a gravidade.(AU)

Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis

High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of High mobility group (HMG) family, play important role in inflammation. The purposes of this study were to investigate the expression of HMGB1 and HMGN2 in periodontistis. The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1, IL-6, IL-8, TNF- and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA. HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1, IL-6, IL-8 proinflammaory cytokines. To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.

Expressão gênica de TLR-2, TLR-4, HMGB1 e VEGF em úlcera abomasal em bovinos de corte

As úlceras abomasais atingem bovinos de todas as idades e raças em todos os sistemas de produção, gerando perdas econômicas. A úlcera resulta da isquemia, atraindo leucócitos e macrófagos, estimulando fibroblastos, células endoteliais e epiteliais. A proteína do grupo de alta mobilidade 1 (HMGB1) liga -se a diferentes receptores de superfície celular, incluindo Toll-like-2 (TLR-2) e -4 (TLR-4), produzindo citocinas. A presença da HMGB1 causa aumento dos níveis do fator de crescimento endotelial vascular (VEGF), um regulador fundamental da angiogênese. Assim, investigou -se a participação da HMGB1, TLR-2, TLR-4 e VEGF em úlceras abomasais em bovinos de corte. Um total de 150 abomasos de bovinos de corte foi examinado em um abatedouro; 17 amostras da região cárdica foram colhidas. Os tecidos extraídos foram classificados em grupo normal (sem ulceração de mucosa); ulceração de grau 1 (erosões não perfuradas com lesões mínimas da mucosa) e ulceração de grau 2 (erosões não perfuradas combinadas com sangramento moderado da mucosa) e confirmado pela histopatologia. A expressão dos genes nas amostras normais ou ulceradas no abomaso foi avaliada pela RT qPCR. Os dados foram submetidos à ANOVA seguido por teste de Bonferroni ao nível de p<0,05. Não houve diferença de expressão de HMGB1, de TLR-4 e de VEGF entre os dois tipos de úlceras em relação aos abomasos normais. Úlceras de grau 2 tiveram expressão de TLR-2 superior a úlceras de grau 1. O aumento da expressão de TLR-2 pode estar associado à manutenção da cicatrização, promovendo a resposta inflamatória, evidenciado pela presença de infiltrado inflamatório mononuclear e neutrófilos.

Abomasal ulcers affect cattle of all ages and breeds in all production systems, leading to economic losses. The ulcer resulting from tissue ischemia, attracting leukocytes and macrophages, stimulates fibroblasts, endothelial and epithelial cells. The protein of high mobility group 1 (HMGB1) binds to different cell surface receptors, including Toll-like-2 (TLR-2) and 4 (TLR-4) resulting in cytokine production. The presence of HMGB1 causes increased levels of vascular endothelial growth factor (VEGF), a key regulator of angiogenesis. Thus, it was investigated whether HMGB1, TLR-2, TLR-4 and VEGF play a role in abomasal ulcers in beef cattle. A total of 150 abomasums from beef cattle were examined in a slaughterhouse; 17 samples were collected from the cardiac region. The extracted tissues were divided into normal group (without ulceration of the mucosa); type 1 ulcers (unperforated erosions with minimal mucosal injury) and type 2 ulcers (unperforated erosions combined with moderate bleeding of the mucosa) and confirmed by histopathology. Gene expression was evaluated by RT qPCR in samples of normal or ulcerated abomasums. Data were analyzed by ANOVA followed by Bonferroni test at p <0.05. No difference in expression of HMGB1, TLR-4 and VEGF was detected between the two types of ulcers when compared to normal abomasums. TLR-2 expression was higher in type 2 ulcers than in type 1 ulcers. Increased TLR-2 expression might be associated with the maintenance of abomasal healing, promoting the inflammatory response, as evidenced by the presence of mononuclear cell infiltration and neutrophils.