Análise da expressão de mRNA de amostras FFPE de feridas lombares de equinos tratados com óleo de copaíba 10 %
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Data
2022-02-10
Autores
Poló, Tatiane da Silva
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo desta pesquisa foi analisar o transcriptoma de mRNA de feridas cutâneas experimentais na região lombar de equinos tratadas ou não com óleo de copaíba a 10% a partir de biopsia de amostras, fixadas em formalina e embebidas em parafina (FFPE). Foram utilizadas biopsias de feridas experimentais em região lombar de equinos tratadas ou não (controle) com óleo de copaíba a 10% obtidas e armazenadas em FFPE há sete anos. A extração do RNA foi realizada utilizando Kit Qiagen RNeasy FFPE, devidamente quantificado e qualificado para análise da expressão do transcriptoma no GeneChip™ Equine Gene 1.1 ST Array Strip, escaneadas pelo GeneAtlas (Affymetrix®), utilizando o software Transcriptome Analysis Console (TAC) (Affymetrix®) para identificação, empregando-se a análise estatística ANOVA (fold change ± 1,5; FDR corrigido P < 0,05). A metodologia de extração de mRNA mostrou-se eficaz na avaliação de amostras de tecido tegumentar FFPE para realização de transcriptoma. O grupo que recebeu o óleo de copaíba a 10% apresentou cinco mRNA supra regulados, relacionados aos genes MIR365-2, MIR551B , LOC100060487, RPL12 e CNN3, além de três mRNA infra relulados, estes relacionados aos genes LOC102149005, LOC102149966 e C10H19orf33. Este é o primeiro estudo relacionado ao transcriptoma de mRNA de feridas lombares em equinos tratados com óleo de copaíba 10%. Os cinco genes supra regulados estão relacionados ao controle de tecido de granulação, apoptose e migração fibroblástica e, entre os três genes infra regulados, há o envolvimento da proteína ligadora de histonas, que ao ser inibida há diminuição da fibrose em pele, melhorando as características de cicatrização feridas ferida.
The objective of this research was to analyze the mRNA transcriptome of experimental cutaneous wounds in the lumbar region of horses treated or not with 10% copaiba oil from biopsy samples, fixed in formalin and embedded in paraffin (FFPE). Were used biopsies of experimental skin wounds in the lumbar region of horses treated or not (control) with 10% copaiba oil, fixed in formalin and embedded in paraffin (FFPE), obtained and stored seven years ago. The RNA extraction was performed Quiagen RNeasy FFPE kit, properly quantified and qualified for analysis of transcriptome expression in the GeneChip™ Equine Gene 1.1 ST Array Strip, scanted by GeneAtlas (Affymetrix®), and genes identified by Transcriptome Analysis Console (TAC) (Affymetrix® software), using the statistical analysis ANOVA (fold change ± 1.5; FDR corrected p < 0.05). The mRNA extraction methodology was favorable in the evaluation of FFPE samples for transcriptome performance. Comparing the groups, 10% copaiba oil showed increased expression of five mRNAs related to genes MIR365-2, MIR551B, LOC100060487, RPL12 and CNN3, and decreased expression of three mRNAs related to genes LOC102149005, LOC102149966 and C10H19orf33. This is the first study related to mRNA transcriptome from lumbar wounds in horses treated with 10% copaiba oil of healing wounds. The five genes upregulated are related to the control of granulation tissue, apoptosis and fibroblast migration and, among the three downregulated genes, there is the involvement of the histone-binding protein, which, when inhibited, reduces fibrosis in the skin, improving the characteristics of the skin.
The objective of this research was to analyze the mRNA transcriptome of experimental cutaneous wounds in the lumbar region of horses treated or not with 10% copaiba oil from biopsy samples, fixed in formalin and embedded in paraffin (FFPE). Were used biopsies of experimental skin wounds in the lumbar region of horses treated or not (control) with 10% copaiba oil, fixed in formalin and embedded in paraffin (FFPE), obtained and stored seven years ago. The RNA extraction was performed Quiagen RNeasy FFPE kit, properly quantified and qualified for analysis of transcriptome expression in the GeneChip™ Equine Gene 1.1 ST Array Strip, scanted by GeneAtlas (Affymetrix®), and genes identified by Transcriptome Analysis Console (TAC) (Affymetrix® software), using the statistical analysis ANOVA (fold change ± 1.5; FDR corrected p < 0.05). The mRNA extraction methodology was favorable in the evaluation of FFPE samples for transcriptome performance. Comparing the groups, 10% copaiba oil showed increased expression of five mRNAs related to genes MIR365-2, MIR551B, LOC100060487, RPL12 and CNN3, and decreased expression of three mRNAs related to genes LOC102149005, LOC102149966 and C10H19orf33. This is the first study related to mRNA transcriptome from lumbar wounds in horses treated with 10% copaiba oil of healing wounds. The five genes upregulated are related to the control of granulation tissue, apoptosis and fibroblast migration and, among the three downregulated genes, there is the involvement of the histone-binding protein, which, when inhibited, reduces fibrosis in the skin, improving the characteristics of the skin.
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Palavras-chave
Cavalo, Fitoterapia, Transcriptoma, Horses, Phytotherapy, Transcriptome