A simple, easy, and safe procedure aiming to improve liver regeneration could be of great clinical benefit in critical situations such as major hepatectomy, trauma, or hemorrhage. Low-power laser irradiation (LPLI) has come into a wide range of use in clinical practice by inducing regeneration in healthy and injured tissues. However, the effect of LPLI on the process of liver regeneration, especially those related to the molecular mechanisms, is not fully understood. Thus, the aim of the present study was to investigate the main molecular mechanisms involved in liver regeneration of partially hepatectomized rats exposed to LPLI. We used Wistar male rats, which had their remaining liver irradiated or not with LPLI (wavelength of 632.8 nm and fluence of 65 mW/cm(2)) for 15 min after a 70% hepatectomy. We subsequently investigated hepatocyte growth factor (HGF), Met, Akt, and Erk 1/2 signaling pathways through protein expression and phosphorylation analyses along with cell proliferation (proliferating cell nuclear antigen (PCNA) and Ki-67) using immunoblotting and histological studies. Our results show that LPLI can improve liver regeneration as shown by increased HGF protein expression and the phosphorylation levels of Met, Akt, and Erk 1/2 accompanied by higher levels of the PCNA and Ki-67 protein in the remnant livers. In summary, our results suggest that LPLI may play a clinical role as a simple, fast, and easy-to-perform strategy in order to enhance the liver regenerative capacity of a small liver remnant after hepatectomy.

The liver regeneration is an important clinical issue after major hepatectomies. Unfortunately, many organs (including the liver) exhibit age-related impairments regarding their regenerative capacity. Recent studies found that low-power laser irradiation (LPLI) has a stimulatory effect on the liver regeneration process. However, its effects in elderly remain unknown. Thus, this study aimed to investigate the main molecular mechanisms involved in liver regeneration of partially hepatectomized elderly rats exposed to LPLI. The effects of 15 min of LPLI (wavelength of 632.8 nm; fluence of 0.97 J/cm(2); total energy delivered of 3.6 J) were evaluated in hepatectomized elderly Wistar male rats. Afterwards, through immunoblotting approaches, the protein expression and phosphorylation levels of hepatocyte growth factor (HGF), Met, Akt and Erk 1/2 signaling pathways as well as the proliferating cell nuclear antigen (PCNA) were investigated. It was observed that LPLI was not able to improve liver regeneration in elderly rats as evidenced by the lack of improvement of HGF and PCNA protein expression or phosphorylation levels of Met, Akt and Erk 1/2 in the remnant livers. In sum, this study demonstrated that the main molecular pathway, i.e. HGF/Met → Akt and Erk 1/2 → PCNA, involved in the hepatic regeneration process was not improved by LPLI in elderly hepatectomized rats, which in turn rules out LPLI as an adjuvant therapy, as observed in this protocol of liver regeneration evaluation (i.e. at 48 h after 70 % resection), in elderly.

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Protoporphyrin IX (PpIX) is a precursor of heme synthesis and is known to be an active photosensitizer and precursor of photosensitizers applied in photodynamic therapy (PDT) and photodynamic diagnostics (PDD). On irradiation with visible light, PpIX undergoes phototransformation, producing photoproducts which may also be phototoxic and increase its efficacy. The mechanism of PpIX phototransformation depends on environmental characteristics and can be different in vitro and in vivo. In this paper, we present a comparative study of the photoactivity of synthetic PpIX and PpIX extracted from the Harderian gland of ssp Rattus novergicus albinus rats, along with their photoproducts toward murine B16F-10 melanoma cells. It was observed that when irradiated with visible light the endogenous PpIX demonstrates photocytotoxicity ten times higher than the synthetic PpIX. The photoproduct of endogenous PpIX also possesses phototoxicity, though slightly lower than that of PpIX itself. The rate of cell internalization for both endogenous PpIX and its photoproduct was eightfold greater than that obtained for the synthetic porphyrin. This difference might result from a complexation of the native PpIX with some amphiphilic compounds during its synthesis within the Harderian glands, which facilitates the cell uptake of PpIX. Fluorescence microscopy images show that both endogenous and synthetic porphyrins are localized after uptake predominantly in the mitochondrial region of cells.

Purpose: To show that a non-laser light source can be constructed, using a 500 W Tungsten lamp and optical filters, and that this light source produces photodynamic effect via protoporphyrin IX (PpIX) similar to the effect produced by HeNe laser light. Methods: The broad band spectrum from a Tungsten lamp was filtered. Infrared and blue part of the spectrum was discarded by absorption process and the fraction of the spectrum, centered at the red portion, was filtered by an interference filter. Photodynamic effect was studied by the activity on endogenous PpIX of Harderian glands of Wistar rats. Twenty rats were used for the experiment. Each animal had its two Harderian gland surgically exposed, so one of them was treated with the system while the other was kept as control. After a 30 minutes period of treatment, the animals were sacrificed and their glands were removed for histological analysis. This analysis was compared to earlier published results obtained with HeNe laser light. Results: The resultant light source emission was centered around (636 ± 6.5) nm and gives up to 11.3 mW/cm2 power density. It produces photodynamic effect in Harderian gland, observed either by fluorescence spectroscopy or by histological microscopy. Conclusion: There is no noticeable difference in Photodynamic effect results if activated by HeNe laser or by the proposed non-laser light source emitting at the red portion of the spectrum. Key words: Photodynamic Therapy. Lasers. Harderian gland. RESUMOObjetivo: Descrever a construção de uma fonte de luz não-laser, a partir de uma lâmpada de Tungstênio e filtros óticos adequados e demonstrar que sua eficiência em estudos fotodinâmicos, mediados por protoporfirina IX, é semelhante a do laser de Hélio Neônio. Métodos: As regiões Infravermelha e Azul do espectro de emissão óptica de uma lâmpada de Tungstênio foram convenientemente descartadas por processos de absorção, enquanto que a fração centrada na região do vermelho foi removida com o uso de um filtro de interferência. O efeito fotodinâmico foi estudado, em glândula Harderiana de ratos Wistar em razão da produção endógena de protoporfirina IX (PpIX) por estas glândulas. Foram utilizados 20 ratos. Cada animal teve as duas glândulas expostas cirurgicamente, sendo uma delas tratada com a fonte não-laser e a outra mantida como controle. Após tratamento por 30 minutos os animais foram sacrificados e suas glândulas removidas para estudo histológico. Os resultados foram comparados a estudos realizados com laser de Hélio Neônio, já publicados. Resultados: A luz produzida pelo equipamento está centrada em torno de (636 ± 6,5) nm, fornecendo uma densidade de potência de 11,3 mW/cm2. Os efeitos fotodinâmicos produzidos na glândula Harderiana, podem ser observados tanto por espectroscopia de fluorescência como por microscopia ótica. Conclusão: Não foram observadas diferenças significativas nos resultados do efeito fotodinâmico obtidos com a fonte de luz não-laser proposta, em comparação aos resultados conhecidos com o uso do laser de Héli...