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Objective To investigate the intervention effect of GDC-0449, a hedgehog signaling pathway inhibitor, on rats with liver fibrosis induced by carbon tetrachloride (CCl 4 ) combined with 2-acetylaminofluorene (2-AAF). Methods A total of 18 female Fisher344 rats were randomly divided into normal group, CCl 4 /2-AAF group, and GDC-0449 group, with 6 rats in each group. The rats in the CCl 4 /2-AAF group and the GDC-0449 group were given subcutaneously injected 30% CCl 4 -olive oil solution at a dose of 2 mL/kg twice a week for 6 weeks to induce liver fibrosis; since week 7, in addition to the injection of CCl 4 -olive oil solution, the rats in these two groups were given 2-AAF (100 mg/kg/d) by gavage, and the rats in the GDC-0449 group were given GDC-0449 (25 mg/kg/d) by gavage, while those in the normal group were given an equal volume of olive oil solution by injection and normal saline by gavage. All rats were sacrificed at the end of week 9, and related samples were collected. HE staining and sirius red (SR) staining were used to observe the changes in liver histopathology and collagen deposition, and the semi-quantitative analysis of SR-positive area and Ishak score were used to evaluate fibrosis degree; the alkaline hydrolysis method was used to measure the level of hydroxyproline (Hyp) in liver tissue; immunohistochemistry, Western blot, and qRT-PCR were used to measure the expression of α-smooth muscle actin (α-SMA), type Ⅰ collagen (Col-Ⅰ), type Ⅳ collagen (Col-Ⅳ), cytokeratin 19 (CK19), cytokeratin 7 (CK7), the epithelial cell adhesion molecule Epcam, and the hedgehog signaling pathway in liver tissue; double immunofluorescence staining was used to observe the colocalization of CK19 and the oval cell marker OV6. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the normal group, the CCl 4 /2-AAF group had marked inflammatory cell aggregation and collagen deposition in liver tissue, with the formation of a pseudolobular structure, as well as significant increases in Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Col-Ⅰ, Col-Ⅳ, Epcam, CK19, CK7, the transmembrane transporter Smoothened (Smo), Hedgehog ligand Desert Hedgehog (Dhh), the Indian Hedgehog membrane-binding receptor Patched (Ptch2), and glioma-related oncogenes Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed that CK19-positive cells also expressed OV6 in the liver tissue of rats in the CCl 4 /2-AAF group, with a significant increase compared with the normal group. Compared with the CCl 4 /2-AAF group, the GDC-0449 group had significant reductions in inflammatory cell aggregation and collagen deposition in liver tissue, Hyp level and collagen positive area ratio in liver tissue ( P < 0.05), Ishak score ( P < 0.05), and the expression of α-SMA, Epcam, CK19, CK7, Smo, Ptch2, Gli1, Gli2, and Gli3 (all P < 0.05); double immunofluorescence staining showed a significant reduction in the number of cells with co-expression of OV6 and CK19 in liver tissue. Conclusion The Hedgehog signaling pathway inhibitor GDC-0449 can significantly inhibit the progression of liver fibrosis induced by CCl 4 /2-AAF in rats, possibly by inhibiting hepatic stellate cell activation, collagen deposition, activation and proliferation of hepatic progenitor cells, and differentiation of hepatic progenitor cells into biliary epithelial cells.
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Objective To investigate the effects of nicotine on the gene expression profile in prefrontal cortex (PFC) region of rats. Methods Twenty-four adult male F344 rats were randomly divided into treatment group and control group, with 12 animals in each group. Animals in treatment group were injected with nicotine solution while those in control group were given physiological saline for 17 days. At the end of drug administration, the tissue of PFC region was sliced from the brain of each animal. RNA sequencing (RNA-Seq) was utilized to analyze the gene expression profile in PFC of each animal. Bioinformatics tools including Bowtie, Tophat and Cufflinks were used to map the sequencing reads to the reference genome of rat, and to measure the relative expression level of each transcript. Genes whose expression levels were significantly differ-ent between the two groups were identified. The functional categories and the biochemical pathways associated with these genes were further analyzed. Results By comparing the expression level of each transcript between two groups, 885 signifi-cantly differentially expressed genes were identified. These genes were enriched in multiple Gene Ontology Biological Pro-cess categories (e.g., G protein coupled receptor protein signaling pathway, protein ubiquitination, and neurotransmitter up-take) and biological pathways (e.g., insulin signaling pathway, Alzheimer's disease, and long term potentiation). Conclu-sion Nicotine treatment may regulate signaling transduction, energy metabolism and other biological processes in PFC of rat brain.
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Objective: The present study assessed the effect of prolonged betamethasone use in ligature-induced periodontitis in adult Fischer-344 rats. Methods: Thirty-six Fisher rats were randomly assigned to three groups: group B (betamethasone); group S (sham) and group C (control). The animals in group B were given intramuscular betamethasone daily, those in group S were given saline injections daily, and those in group C were not submitted to any intervention. The interventions lasted 60 days, and all groups were submitted to the same environmental conditions. Tendays after starting the injections, periodontal disease was induced in the animals from groups B and S by tying a sterile silk suture threadaround the upper right second molar. Fifty days later, all animals were sacrificed and the connective tissue attachment level (Jac-E) and boneloss (Jac-O) were measured. Results: Jac-E and Jac-O of groups B and S were not significantly different, but they differed significantly from those of group C. Conclusion: Prolonged use of betamethasone did not affect the progression of induced periodontitis in rats.
Objetivo: Avaliar o efeito do uso prolongado da betametasona na periodontite induzida por ligadura, em ratas adultas da linhagem Fischer-344. Métodos: Selecionou-se trinta e seis ratas Fisher, as quais foram divididas aleatoriamente em três grupos: grupo B (betametasona); grupo S (soro) e grupo C (controle). Os animais do grupo B receberam injeções diárias intramuscular de betametasona, o grupo S recebeu injeções diárias de soro fisiológico e o grupo C foi mantido sem nenhuma intervenção. Estes procedimentos duraram 60 dias. Os três grupos foram mantidos nas mesmas condições ambientais. Decorridos dez dias a partir do início das injeções, os animais dos grupos B e S foram submetidos a indução da doença periodontal, através da colocação do fio de seda em volta do segundo molar superior direito. Ao final do período experimental (50 dias) os animais foram submetidos à eutanásia. O nível de inserção histológica (Jac-E) e perda óssea histológica (Jac-O) foram mensurados.Resultados: Entre os grupos B e S, não houve diferenças estatisticamente significativas para Jac-E e Jac-O. Entretanto, notaram-se diferenças significativas em ambos os parâmetros avaliados quando o grupo C foi comparado aos grupos B e S. Conclusão: Pode-se concluir que o uso prolongado de betametasona não modificou a progressão de periodontite induzida.
Subject(s)
Animals , Betamethasone , PeriodontitisABSTRACT
Objetivo: Compreender o efeito do uso crônico de álcool na progressão de periodontite induzida em ratos da linhagem Fischer-344. Métodos: Para o estudo utilizaram-se 22 ratos da linhagem Fischer-344, com dois meses de idade, divididos nos grupos: álcool (n=8), ligadura (n=7) e controle (n=7). No primeiro dia, expuseram-se os animais do grupo álcool à ingestão de solução de água com álcool a 20% (volume/volume), até o dia 90. Decorridos trinta dias do início do experimento, os animais do grupo álcool e do grupo ligadura submeteram-se à colocação de fio de seda em volta do segundo molar superior direito. Nada foi realizado no lado esquerdo, servindo como controle. Todos os grupos submeteram-se à eutanásia, após 60 dias da colocação da ligadura. Para avaliar a destruição da periodontite, usou-se o exame radiográfico, mensurando a destruição da altura óssea. Resultados: Os resultados do estudo evidenciaram que, no lado em que foi induzida a periodontite, o grupo que ingeriu álcool sofreu um aumento de destruição, demonstrando diferenças estatísticas (p < 0,05), se comparado ao grupo ligadura e ao controle. E, maior destruição óssea no grupo ligadura quando comparado ao controle (p < 0,05). Conclusão: Conclui-se, dentro das limitações do estudo, que o consumo crônico de álcool por ratos da linhagem Fischer-344 induziu a uma maior progressão de periodontite induzida.
Objective: Understand the effect of chronic alcohol on the progression of periodontitis induced in Fischer-344 rats. Methods: For the study, 22 Fischer-344 rats, two months old were used, divided into groups: alcohol (n=8), ligature (n=7) and control (n=7). On the first day, the animals in the alcohol group were exposed to ingestion of a water solution containing 20% alcohol (size/size), up to day 90. After thirty days from the beginning of the experiment, the animals in the alcohol group and the ligature group were submitted to the placement of a silk thread around the right maxillary second molar. Nothing was performed on the left side, serving as control. All the groups were submitted to euthanasia 60 days after ligature placement. To assess the destruction of periodontitis, a radiographic exam was used to measure the destruction of bone height. Results: The results of the study showed that on the side in which periodontitis was induced, the group that ingested alcohol suffered an increase in destruction, with statistical differences when compared with the ligature and control groups and increased bone destruction in the ligature group when compared to control. Conclusion: Within the limitations of the study, it was concluded that chronic alcohol consumption by Fischer-344 rats led to greater progression of induced periodontitis.
Subject(s)
Animals , Rats , Ethanol/adverse effects , Periodontitis/chemically induced , Case-Control Studies , Disease Models, AnimalABSTRACT
Objective Dynamical observation the development law of rejection of small intestinal transplantation(SIT) in inbred rats BN to F334,to study the value of this SIT madel in the rsesarch of rejection. Methods Heterotopic SIT models were established by microsurgical technique according to modified monchik method in inbred strain F344/RT1 l andBN/RT1 n,including isotransplantation (F344→F344, n =8 ,ITx group) and allotransplantation (BN→F344, n =8, ATx group). Results (1) The mean survival time was over 30 days in AT X group, 12 days in IT X group(P