| Introduction: Traditional Chinese Medicine(TCM) is the result of long-term practice, and it has been proved that TCM is effective. But the active components and molecular mechanism for TCM were generally poorly understood because of their complex components. Thus, it is important to understand the mechanism for TCM. TCM is usually thought to act on multiple targets with complex mechanism. We can screen some active components of TCM if we study its molecular mechanism by separating and analyzing the chemical components for TCM using the method applied in the research of chemical drugs. However, genes in vivo do not act by themselves, but act through complex gene network. Thus, We should explore the molecular mechanism for TCM at gene network levels, which provides the possible methods for verifying the theory of TCM. DNA chip technology can rapidly, massively and parallelly monitor and analyze a large number of gene expressions. It will play an important role in TCM modernization to establish DNA microarray technology platform which aims to explore the molecular mechanism for TCM.Experimental focal glomerular sclerosis (FGS) can be induced by both immunologic and nonimrnunologic methods, As an experimental analogue of human focal glomerule sclerosis, Previously, adriamycin (ADR)-induced a stable and reproducible murine model of chronic progressive nephropathy with significant and persistent proteinuria has been established in BALB/c mice. On the other hand, Interstitial inflammatory cell infiltration is an obvious event in this model, then. Accumulation and activation of Macrophages and T cells prouce many cytokines, such as interleukin-1, interleukin-6, tumor necrosis facter-a ,platelet-derived growth factor, and transforming growth factor- β , and The cytokines play an important role in glomerular injury and stimulation of mesangial cell proliferation and matrix formation . Moreover, In previous study we have found the TCM of Compound cat'foot has a well useon renal disease.Objective: In order to study the cytokine gene expression profile in mice FGS model and the mechanism of Compound cat'foot on cytokines genes . Methods: (1) 74 cytokines genes genes microarray was prepared, The key technique and standard of quality controlling for preparation of oligonucleotide microarray were explored using human 23KD highly basic protein gene and luciferase gene and 74 cytokines genes. By using of a self-made software system, oligonucleotide probes were designed and Blast. All the probes have a very high specificity, i.e. except target sequence, the similarity between the probe and non-target sequences is less than 70%. In the probes with the same length 40, GC content are in a narrow scope (from 45% to 55%). All the probes are modified with amino at 5'or 3'terminal.The optimal concentration is 0.5ug/ul. (2) The cytokines genes microarray was applied to study the mechanism of Compound cat'foot in mice FGS model induced by ADR. Results: The satisfied images with good sensitivity were obtained by using the methods above and also using positive and negative controls and some internal controls to quantify and balance expression of genes. Good sensitivity and stability have been verified by three continuous experiments using the oligonucleotide microarray to study gene expression profile of kidney tissue. The oligonucleotide microarray for Expression Detection prepared using our method have good sensitivity and stability. It may be a more advanced method for analysis of gene expression profile.The results of DNA microarry examination indicate:①The expression of PDGF- α , PDGF- β , PDGF-R and VEGF mRNA were down-regulated after treated by Compound cat'foot', ② The expression of IL-9, IL-7R2, MIP et al were down-regulated after treated by Compound cat'foot; ;③At the same time, through the stimulation of Compound cat'foot , The expression of c-Myc and Jun in kidney were restrained. It follows that Compound cat'foot enhance immunity system and treat renal diseases related to these cytokines. Conclusion:... |
PDF Full Text Request