| ObjectiveTo find penitential target genes of local oxidative phosphorylation at ST36influenced by moxibustion through genomic microarray and bioinformatics tools, then using quantitative real-time PCR to detect the expression of target genes. Our study may provid new evidences for the research of the response of acupoint to moxibustion from the point of oxidative phosphorylation.Methods(1) Looking for the pathway of oxidative phosphorylation and it’s related genes at ST36(Zusanli) after moxibustion:Twenty-four Sprague-Dawley rats were randomly assigned to control group, model group, mild moxibustion, moxibustion with monkshood cake insulation group. Rats excepting in control group were injected Freund’s complete adjuvant into right foot pad to induce adjuvant arthritis (AA) model. Seven days after injection, mild moxibustion and moxibustion with monkshood cake insulation group were performed at ST36respectively, two hours after treatment, tissues around ST36acupoint were took immediately. Total RNA of each sample was extracted and purified. The sample and the rat common reference were labeled by flouresence with cRNA in vitro transcription and Linear amplification. We employed27k RNAS Oligo Microarrays to detected the gene expression profiles of ST36tissues and to screen differentially expressed genes by SAM(Significance analysis of microarrays) software, and we used Random Variance Model and MAS(Molecule annotation system) software to select the pathway of co-expression genes.(2) biology-repeated experiment:Rats were accepted the same operation as the first experiment, then we used quantitative real-time PCR immediately to detect the expression of target genes after extracting the total RNA. Results1.498different expressed genes (DEGs) were detected after mild moxibustion,255DEGs belong to up-regulated genes and243DEGs belong to down-regulated genes.937DEGs were detected after moxibustion with monkshood cake insulation,355DEGs belong to up-regulated genes and602DEGs belong to down-regulated genes.106co-expression DEGs were detected,44co-expression DEGs belong to up-regulated genes and62co-expression DEGs belong to down-regulated genes.106co-expression DEGs were obviously took part in27pathway (P<0.001), Cox7a2, Atp6v0d2and Atp5j were took part in the pathway oxidative phosphorylation. We use foldchange≥2.0&|Score(d)|≥2and foldchange<0.5&|Score(d)|≥2as as our standard to select up-regulated genes and down-regulated genes.2. The result of biology-repeated experiment as follow:Compared to those in control group, the expression of Cox7a2, Atp6v0d2and Atp5j in model group were significantly down-expressed(.P<0.05). Compared to the model group, both of mild moxibustion group and moxibustion with monkshood cake insulation group increased expression of Cox7a2, Atp6v0d2and Atp5j (P<0.05), the results are consistent with the results of gene chip. Furthermore, moxibustion with monkshood cake insulation showed more significant influence on the expression of the Cox7a2, Atp6v0d2and Atp5j (P<0.05)ConclusionsThe pathway of oxidative phosphorylation may take part in the response of acupoint to moxibustion, Cox7a2, Atp6v0d2and Atp5j may be the penitential target genes of oxidative phosphorylation pathway at ST36after moxibustion. |
PDF Full Text Request