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1.
Acta cir. bras ; 31(8): 505-512, Aug. 2016. tab
Artigo em Inglês | LILACS | ID: lil-792408

RESUMO

ABSTRACT PURPOSE: To evaluate the effect of keratinocyte growth factor (KGF) treatment on the expression of wound-healing-related genes in cultured keratinocytes from burn patients. METHODS: Keratinocytes were cultured and divided into 4 groups (n=4 in each group): TKB (KGF-treated keratinocytes from burn patients), UKB (untreated keratinocytes from burn patients), TKC (KGF-treated keratinocytes from controls), and UKC (untreated keratinocytes from controls). Gene expression analysis using quantitative polymerase chain reaction (qPCR) array was performed to compare (1) TKC versus UKC, (2) UKB versus UKC, (3) TKB versus UKC, (4) TKB versus UKB, (5) TKB versus TKC, and (6) UKB versus TKC. RESULTS: Comparison 1 showed one down-regulated and one up-regulated gene; comparisons 2 and 3 resulted in the same five down-regulated genes; comparison 4 had no significant difference in relative gene expression; comparison 5 showed 26 down-regulated and 7 up-regulated genes; and comparison 6 showed 25 down-regulated and 11 up-regulated genes. CONCLUSION: There was no differential expression of wound-healing-related genes in cultured primary keratinocytes from burn patients treated with keratinocyte growth factor.


Assuntos
Humanos , Animais , Masculino , Feminino , Adulto , Camundongos , Cicatrização/genética , Queimaduras/genética , Expressão Gênica/genética , Queratinócitos/efeitos dos fármacos , Fator 7 de Crescimento de Fibroblastos/farmacologia , Pele/citologia , Queimaduras/patologia , Estudos de Casos e Controles , Regulação para Baixo , Células Cultivadas , Reação em Cadeia da Polimerase
2.
Clinics ; 71(8): 470-473, Aug. 2016. tab
Artigo em Inglês | LILACS | ID: lil-794636

RESUMO

OBJECTIVE: The aim of this study was to evaluate the roles of the Taql and Bsml vitamin D receptor gene polymorphisms in hospital mortality of burn patients. METHODS: In total, 105 consecutive burn injury patients over 18 years in age who were admitted to the Burn Unit of Bauru State Hospital from January to December 2013 were prospectively evaluated. Upon admission, patient demographic information was recorded and a blood sample was taken for biochemical analysis to identify the presence of the Taql(rs731236) and Bsml(rs1544410) polymorphisms. All of the patients were followed over their hospital stay and mortality was recorded. RESULTS: Eighteen of the patients did not sign the informed consent form, and there were technical problems with genotype analysis for 7 of the patients. Thus, 80 patients (mean age, 42.5±16.1 years) were included in the final analysis. In total, 60% of the patients were male, and 16.3% died during the hospital stay. The genotype frequencies for the Taql polymorphism were 51.25% TT, 41.25% TC and 7.50% CC; for the Bsml polymorphism, they were 51.25% GG, 42.50% GA and 6.25% AA. In logistic regression analysis, after adjustments for age, gender and total body surface burn area, there were no associations between the Taql (OR: 1.575; CI95%: 0.148-16.745; p=0.706) or Bsml (OR: 1.309; CI95%: 0.128-13.430; p=0.821) polymorphisms and mortality for the burn patients. CONCLUSIONS: Our results suggest that the Taql and Bsml vitamin D receptor gene polymorphisms are not associated with hospital mortality of burn patients.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Polimorfismo Genético , Queimaduras/genética , Queimaduras/mortalidade , Mortalidade Hospitalar , Receptores de Calcitriol/genética , Potássio/sangue , Sódio/sangue , Ureia/sangue , Albumina Sérica , Modelos Logísticos , Análise Multivariada , Estudos Prospectivos , Fatores de Risco , Creatinina/sangue , Genótipo , Tempo de Internação
3.
Acta cir. bras ; 29(supl.3): 33-38, 2014. graf
Artigo em Inglês | LILACS | ID: lil-726241

RESUMO

PURPOSE: To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns. METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences). RESULTS: After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0). CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome. .


Assuntos
Adulto , Feminino , Humanos , Masculino , Queimaduras/genética , Expressão Gênica , Queratinócitos/metabolismo , Receptores Toll-Like/genética , Células Cultivadas , Epiderme/lesões , Epiderme/metabolismo , Reação em Cadeia da Polimerase , RNA , Receptores Toll-Like/metabolismo , Cicatrização
4.
Acta cir. bras ; 29(supl.3): 39-43, 2014. graf
Artigo em Inglês | LILACS | ID: lil-726244

RESUMO

PURPOSE: To evaluate KGF and human beta defensin-4 (HBD-4) levels produced by dermic fibroblasts and keratinocytes cultivated from burned patients' skin samples. METHODS: Keratinocytes and fibroblasts of 10 patients (four major burns, four minor burns and two controls) were primarily cultivated according to standard methods. HBD-4 and KGF genes were analyzed by quantitative PCR. RESULTS: In fibroblasts, KGF gene expression was 220±80 and 33.33±6.67 (M±SD; N=4), respectively for major and minor burn groups. In keratinocytes, KGF gene expression was 11.2±1.9 and 3.45±0.37 (M±SD; N=4), respectively for major and minor burn groups. In fibroblasts, HBD-4 gene expression was 15.0±4.0 and 11.5±0.5 (M±SD; N=4), respectively for major and minor burn. In keratinocyte, HBD-4 gene expression was 0.0±0.0 and 13.4±4.8 (M±SD; N=4), respectively for major and minor burn. CONCLUSIONS: KGF expression was increased in burn patient fibroblasts compared to control group. In keratinocytes culture, KGF suppression is inversely proportional to burn extension; it is active and increased in major burn but decreased in minor burn. HBD-4 expression was increased in fibroblasts and decreased in keratinocytes from all burned patients. .


Assuntos
Feminino , Humanos , Masculino , Adulto Jovem , Queimaduras/genética , /análise , Fibroblastos/metabolismo , Queratinócitos/metabolismo , beta-Defensinas/genética , Células Cultivadas , /genética , Expressão Gênica , Reação em Cadeia da Polimerase , RNA , Pele/citologia , Pele/lesões , beta-Defensinas/metabolismo
5.
Acta cir. bras ; 29(supl.3): 60-67, 2014. graf
Artigo em Inglês | LILACS | ID: lil-726247

RESUMO

PURPOSE: Evaluate the expression profile of genes related to Innate and Adaptive Immune System (IAIS) of human Primary Epidermal keratinocytes (hPEKP) of patients with severe burns. METHODS: After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific IAIS PCR Arrays plates (SA Biosciences). RESULTS: After the analysis of gene expression we found that 63% of these genes were differentially expressed, of which 77% were repressed and 23% were hyper-regulated. Among these, the following genes (fold increase or decrease): IL8 (41), IL6 (32), TNF (-92), HLA-E (-86), LYS (-74), CCR6 (- 73), CD86 (-41) and HLA-A (-35). CONCLUSIONS: This study contributes to the understanding of the molecular mechanisms underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome. .


Assuntos
Adulto , Feminino , Humanos , Masculino , Imunidade Adaptativa/genética , Queimaduras/genética , Expressão Gênica , Imunidade Inata/genética , Queratinócitos/citologia , Imunidade Adaptativa/imunologia , Queimaduras/imunologia , Células Cultivadas , Queratinócitos/imunologia , Reação em Cadeia da Polimerase , Projetos de Pesquisa , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/genética , Cicatrização/genética
6.
Acta cir. bras ; 28(8): 551-558, Aug. 2013. graf, tab
Artigo em Inglês | LILACS | ID: lil-680608

RESUMO

PURPOSE: To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. METHODS: Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. RESULTS: The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). CONCLUSIONS: The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.


Assuntos
Adulto , Feminino , Humanos , Masculino , Queimaduras/genética , /genética , Fibroblastos/metabolismo , Interleucina-1beta/genética , Fator de Necrose Tumoral alfa/genética , Estudos de Casos e Controles , Células Cultivadas , /análise , Expressão Gênica , Interleucina-1beta/análise , Reação em Cadeia da Polimerase , Pele/citologia , Pele/lesões , Fatores de Tempo , Fator de Necrose Tumoral alfa/análise
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