RESUMEN
To investigate molecular mechanism of traditional Chinese medicine Rheum offcinale against Yersinia pestis, whole genome DNA microarray that contains 4005 annotated genes of Y. pestis was used. The minimal inhibition concentration (MIC) of R. offcinale extract against Y. pestis was determined by liquid dilution method. The gene expression profile of Y. pestis was performed after exposured to R. offcinale extract at a concentration of 10 X MIC for 30 and 60 minutes. The total RNA extracted and purified from Y. pestis were reverse-transcribed to cDNA and labeled by Cy3-Cy5 dye. The labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software. The microarray data was confirmed by RT-PCR. The platform of the DNA microarray-based bacteria transcriptional profiling was eshtablished. The results revealed general gene expression changes of Y. pestis were a global phenomenon. Down-regulation of genes encoding proteins involved in ribosome protein synthesis was a remarkable change. Genes encoding cell envelope and transport/binding proteins were the major changed genes of the Y. pestis in response to R. offcinale.
Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Rheum/química , Yersinia pestis/efectos de los fármacos , Proteínas Bacterianas/genética , Regulación hacia Abajo , Perfilación de la Expresión Génica , Pruebas de Sensibilidad Microbiana , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Bacteriano/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rheum/genéticaRESUMEN
Coptis chinensis Franch. is a natural herb widely used in China for prevention and treatment of infectious diseases. Plague is a deadly disease caused by Yersinia pestis. Coptis chinensis Franch. is considered the therapeutic agent of choice against plague rather than conventional antibiotics because of its low cost and low toxicity. Berberine is the major constituent of a Coptis chinensis Franch. extract. In the present study, DNA microarray was used to investigate the transcription of Y. pestis in response to berberine. The minimal inhibition concentration (MIC) of berberine to Y. pestis was determined with the liquid dilution method. The gene expression profile of Y. pestis was performed by exposing Y. pestis to berberine at a concentration of 10 x MIC for 30 min. Total RNA was extracted and purified from Y. pestis, reverse-transcribed to cDNA, and then labeled with Cy-dye probes. The labeled probes were hybridized to the microarray. The results were obtained by a laser scanner and analyzed with SAM software. A total of 360 genes were differentially expressed in response to berberine: 333 genes were upregulated, and 27 were downregulated. The upregulation of genes that encode proteins involved in metabolism was a remarkable change. In addition to a number of genes of unknown encoding or unassigned functions, genes encoding cellular envelope and transport/binding functions represented the majority of the altered genes. A number of genes related to iron uptake were induced. This study revealed global transcriptional changes of Y. pestis in response to berberine, hence providing insights into the mechanisms of Coptis chinensis Franch. against Y. pestis.