ABSTRACT
Kunming mice (male, weight 20 +/- 2g) were daily intragastric administration of Chinese nutgall extract (0.2 mL/10 g body weight, equal to 8 g Chinese nutgall material/1 kg body weight) for 30 days. Liver tissue mRNA were extracted from normal control group and Chinese nutgall treated group respectively, then were reversely transcribed to cDNA with dUTP labeled by different fluorescence (Cy3, Cy5) as hybridization probes. Both of the cDNA probes were mixed equally in 20 microL of hybridization solution and hybridized with mice complete genome oligonucleotide microarray. The fluorescent signals were acquired by laser scanner and analyzed by GenePix Pro 4.0 software. The biological function analysis and the pathway analysis of differentially expressed genes were performed according to Gene Ontology database. As a result, there were 461 genes differentially expressed in Chinese nutgall treated group, in which 373 genes were function-known and the others were function-unknown. Among the 461 genes, 267 genes were up-regulated and the others were down-regulated. The differentially expressed genes were involved in metabolism, DNA binding and transcription, protein synthesis and modification, cell cytoskeleton and cell adhesion, cell cycle and differentiation, ion channels and transporters, signal transduction, immune response and apoptosis of liver cell. The presented work might be quite important for understanding the pathogenic mechanisms of liver injury induced by Chinese nutgall.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Gene Expression Profiling , Liver/drug effects , Oligonucleotide Array Sequence Analysis/methods , Animals , Carbocyanines/pharmacology , Chemical and Drug Induced Liver Injury/etiology , In Situ Hybridization , Liver/metabolism , Male , Mice , Microarray Analysis , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effectsABSTRACT
AIM: To analyze the gene expression profiles of mice livers injured by Leigongteng and explore the relationship between the differentially expressed genes and liver damage. METHODS: The experimental mice were randomly divided into a control group and a liver-injured group in which the mice were administrated 33 mu gamma of triptolide/kg per day for 30 d. Liver mRNAs were extracted from animals in both groups and were reverse-transcribed to cDNA with dUTP labeled by different fluorescence (Cy3, Cy5) as hybridization probes. The mixed probes were hybridized with oligonucleotide microarray chips. The fluorescent signal results were acquired by scanner and analyzed with software. RESULTS: Among the 35852 target genes, 29 genes were found to be significantly differentially expressed, with 20 genes up-regulated and 9 genes down-regulated. The reliability of the differentially expressed genes was validated by RT-PCR experiments of 5 randomly selected differentially expressed genes. CONCLUSION: Based on the biological functions of the differentially expressed genes, it is obvious that the occurrence and development of liver damage induced by Leigongteng in mice are highly associated with immune response, metabolism, apoptosis and the cell skeleton of liver cells. This might be important for elucidating the regulatory network of gene expression associated with liver damage and it may also be important for discovering the pathogenic mechanisms of liver damage induced by Leigongteng.