摘要
AIM: To investigate the effects of four different ingredients of zedoary (Curcuma aromatica oil, Curcumol,β-elemence, and Curcumin) on the gene expressions of hepatic stellate cells (HSCs), and to explore the molecular mechanism of zedoary against hepatic fibrosis at gene network level.METHODS: We detected the mRNA sequences of 50 liver fibrosis-related genes in GenBank and designed oligonucleotide probes. We synthesized oligonucleotides with PE8909 DNA synthesizing instrument, and carried out oligonucleotide microarray with OGR-04 dropping instrument and aldehyded glass chip. Cultured HSC-T6cells were treated with different concentrations of Colchicine,Curcuma aromatica oil, Curcumol, β-elemence, and Curcumin. According to the experiment of cell toxicity,we took the appropriate concentrations of medicines that resulted in over 50% of cell survival as experiment concentrations. We collected the cells at 1, 6, 12, and 24 h, and extracted total RNA with TRIzol reagent, then labeled cDNAs with Cy3-dUTP and Cy5-dUTP. These labeled cDNAs were hybridized to an oligonucleotide microarray which was washed several times and scanned by scanner GenePix 4000B. Different gene expressions of HSC-T6 cells were analyzed by ImaGene 4.2 software.RESULTS: After HSC-T6 cells were cultured in a medium containing 6.25 μg/mL Colchicine for 12 h, expression of TIMP-1 decreased 2.2-folds. After HSC-T6 cells were cultured in a medium containing 78.125 μg/mL of Curcuma aromatica oil for 24 h, the expression of TIMP-2and IL-6 decreased 2.3- and 2.2-folds, respectively.Moreover, after HSC-T6 cells were cultured in a medium containing 1.5625 μg/mL of Curcumol for 12 h, the expression of TGFβ1 and P450a decreased 2.3- and 2.1-folds, respectively.CONCLUSION: Our results may show the possible molecular mechanism of Curcuma aromatica oil and Curcumol against hepatic fibrosis.
AIM: To investigate the effects of four different ingredients of zedoary (Curcuma aromatica oil, Curcumol, β-elemence, and Curcumin) on the gene expressions of hepatic stellate cells (HSCs), and to explore the molecular mechanism of zedoary against hepatic fibrosis at gene network level. METHODS: We detected the mRNA sequences of 50 liver fibrosis-related genes in GenBank and designed oligonucleotide probes. We synthesized oligonucleotides with PE8909 DNA synthesizing instrument, and carried out oligonucleotide microarray with OGR-04 dropping instrument and aldehyded glass chip. Cultured HSC-T6 cells were breated wibh different concentrations of Colchicine, Curcuma aromatica oil, Curcumol, β-elemence, and Curcumin. According to the experiment of cell toxicity, we took the appropriate concentrations of medicines that resulted in over 50% of cell survival as experiment concentrations. We collected the cells at 1, 6, 12, and 24 h, and extracted total RNA with TRIzol reagent, then labeled cDNAs with Cy3-dUTP and Cy5-dUTP. These labeled cDNAs were hybridized to an oligonucleotide microarray which was washed several times and scanned by scanner GenePix 4000B. Different gene expressions of HSC-T6 cells were analyzed by ImaGene 4.2 software. RESULTS: After HSC-T6 cells were cultured in a medium containing 6.25μg/mL Colchicine for 12 h, expression of TIMP-1 decreased 2.2-folds. After HSC-T6 cells were cultured in a medium containing 78.125 μg/mL of Curcuma aromatica oil for 24 h, the expression of TIMP-2 and IL-6 decreased 2.3- and 2.2-folds, respectively. Moreover, after HSC-T6 cells were cultured in a medium containing 1.5625 μg/mL of Curcumol for 12 h, the expression of TGFμ1 and P450a decreased 2.3- and 2.1-folds, respectively. CONCLUSION: Our results may show the possible molecular mechanism of Curcuma aromatica oil and Curcumol against hepatic fibrosis.
基金
Supported by Guandong Traditional Chinese Medicine Bureau No.102135 and Shenzhen Technology Bureau No.200204187,
关键词
HSC-T6细胞
基因表达
莪木
药草
DNA microarray
Curcuma aromatica oil
Curcumol
Hepatic stellate cells
Hepatic fibrosis