O efeito anti-inflamatório e cicatrizante do laser de baixa intensidade na fibrogênese hepática induzida pelo modelo de ligadura do ducto biliar
Data
2020-10-06
Autores
Maehara, Maria Eduarda 
Orientadores
Nagaoka, Márcia Regina
Tipo
Trabalho de conclusão de curso
Título da Revista
ISSN da Revista
Título de Volume
Resumo
A terapêutica do laser de baixa intensidade (LLLT) tem sido extensivamente investigada como um método anti-inflamatório e cicatrizante de feridas. Estudos anteriores revelaram a eficácia do laser de baixa intensidade em órgãos viscerais, entretanto pouco se sabe sobre o seu efeito no fígado. Objetivos: Investigar os efeitos terapêuticos do laser de baixa intensidade na fibrogênese hepática em animais submetidos ao modelo experimental de ligadura do ducto biliar. Metodologia: Ratos machos adultos Wistar (Rattus norvegicus) foram anestesiados e após a laparotomia, o ducto biliar comum foi localizado, isolado e duplamente ligado. Os animais do grupo controle (Sham) passaram pelo mesmo procedimento; no entanto, não tiveram seu ducto biliar duplamente ligado. A terapêutica do laser foi realizada seguindo protocolo de aplicação com intervalos de tempo de 7 dias. A avaliação da inflamação foi feita por dosagem das atividades da aspartato-aminotransferase e alanino-aminotransferase no soro dos ratos. A fibrogênese hepática foi avaliada por análise morfométrica utilizando o microscópio AxioImager A2 (Zeiss, Alemanha) e o processador e analisador de imagem AxioVision versão 4.8 (Zeiss, Alemanha). A morte celular foi avaliada pela atividade hepática de catepsina B e caspase-3 e imuno-histoquímica da caspase-3. A regeneração hepática foi avaliada pela imunodetecção de Ki-67. Nós verificamos que a LLLT por 7 dias aumentou a liberação de glicose, mas não a depuração de BSP nos animais BDL quando comparado com o respectivo BDL controle. Pela análise morfométrica verificamos que LLLT diminuiu o conteúdo de colágeno sem alterar significativamente o conteúdo de parênquima. Interessante, pela análise em microscópio de luz polarizada foi observado que a LLLT altera o tipo de colágeno nos septos fibróticos com predominância de fibras do tipo III. Em relação a morte celular, LLLT levou ao aumento da atividade enzimática da caspase-3, sendo evidenciada pela imuno-histoquímica da caspase-3 maior marcação em hepatócitos. A atividade da catepsina B diminuiu após o tratamento com LLLT por 7 dias. Pela imuno-histoquímica para Ki-67 verificamos que LLLT aumentou a marcação principalmente em células ductulares quando comparado com o grupo BDL controle. Assim, os resultados obtidos até o momento sugerem que a LLLT possa ser alternativa terapêutica não-invasiva para controle e redução da fibrogênese, sendo necessário ainda alguns experimentos para confirmação do efeito benéfico.
The low-level laser therapy has been extensively investigated as an anti-inflammatory and wound healing method. Previous studies have shown the effectiveness of low-level laser therapy in visceral organs; however, little is known about its effect on the liver. Objectives: To investigate the therapeutic effects of low-level laser therapy in the liver fibrogenesis using an experimental model of bile duct ligation. Methods: Adult male Wistar rats (Rattus norvegicus) were anesthetized and after laparotomy, the common bile duct was located, isolated and doubly linked. The control group (Sham) underwent the same procedure; however, it did not have their bile duct doubly connected. Laser therapy was performed following application protocol with time intervals of 7 days. Inflammation was evaluated by serum activity of aspartate aminotransferase and alanine aminotransferase. Hepatic fibrogenesis was analyzed by morphometric analysis using the microscope AxioImager A2 (Zeiss, Germany) and the processor and image analyzer AxioVision version 4.8 (Zeiss, Germany). Liver cell death was evaluated by hepatic activity of cathepsin B and caspase-3 and immunohistochemistry of caspase-3. Liver regeneration was evaluated by Ki-67 immunodetection. We found that LLLT for 7 days increased hepatic glucose release, but not BSP clearance in BDL animals when compared to its respective BDL control. Morphometric analysis showed that LLLT decreased the collagen content without significantly alteration in the parenchyma content. Interestingly, by the polarized light microscope analysis, it was observed that LLLT changes the type of collagen in fibrotic septa with a predominance of type III fibers. In relation to cell death, LLLT increased the enzymatic activity of caspase-3, being evidenced by the caspase-3 immunohistochemistry a greater marking in hepatocytes. Cathepsin B activity decreased after treatment with LLLT for 7 days. By immunohistochemistry for Ki-67 we found that LLLT increased the hepatic regeneration, mainly in ductular cells when compared to the control BDL group. Thus, up to now our results suggest that LLLT may be a non-invasive therapeutic alternative for the control and reduction of fibrogenesis, requiring further experiments to confirm this beneficial effect.
The low-level laser therapy has been extensively investigated as an anti-inflammatory and wound healing method. Previous studies have shown the effectiveness of low-level laser therapy in visceral organs; however, little is known about its effect on the liver. Objectives: To investigate the therapeutic effects of low-level laser therapy in the liver fibrogenesis using an experimental model of bile duct ligation. Methods: Adult male Wistar rats (Rattus norvegicus) were anesthetized and after laparotomy, the common bile duct was located, isolated and doubly linked. The control group (Sham) underwent the same procedure; however, it did not have their bile duct doubly connected. Laser therapy was performed following application protocol with time intervals of 7 days. Inflammation was evaluated by serum activity of aspartate aminotransferase and alanine aminotransferase. Hepatic fibrogenesis was analyzed by morphometric analysis using the microscope AxioImager A2 (Zeiss, Germany) and the processor and image analyzer AxioVision version 4.8 (Zeiss, Germany). Liver cell death was evaluated by hepatic activity of cathepsin B and caspase-3 and immunohistochemistry of caspase-3. Liver regeneration was evaluated by Ki-67 immunodetection. We found that LLLT for 7 days increased hepatic glucose release, but not BSP clearance in BDL animals when compared to its respective BDL control. Morphometric analysis showed that LLLT decreased the collagen content without significantly alteration in the parenchyma content. Interestingly, by the polarized light microscope analysis, it was observed that LLLT changes the type of collagen in fibrotic septa with a predominance of type III fibers. In relation to cell death, LLLT increased the enzymatic activity of caspase-3, being evidenced by the caspase-3 immunohistochemistry a greater marking in hepatocytes. Cathepsin B activity decreased after treatment with LLLT for 7 days. By immunohistochemistry for Ki-67 we found that LLLT increased the hepatic regeneration, mainly in ductular cells when compared to the control BDL group. Thus, up to now our results suggest that LLLT may be a non-invasive therapeutic alternative for the control and reduction of fibrogenesis, requiring further experiments to confirm this beneficial effect.
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Citação
MAEHARA, Maria Eduarda. O efeito anti-inflamatório e cicatrizante do laser de baixa intensidade na fibrogênese hepática induzida pelo modelo de ligadura do ducto biliar. 2020. 42 f. Trabalho de conclusão de curso de graduação (Fisioterapia) - Instituto de Saúde e Sociedade, Universidade Federal de São Paulo, Santos, 2020.