Study On Demethylation Effect Mechanism Of Qinghuang Powder In Treating Myelodysplastic Syndrome

Part One To study effect of Qinghuang power treatment on DNMT1,DNMT3a,DNMT3b mRNA in MDS patientsObjective To study the effect of Qinghuang power on MDS patients’DNMT1, DNMT3a, DNMT3b mRNA expression and observe the correlations between Changes in peripheral blood cells and DNMT1, DNMT3a、DNMT3b. The Research Article focus on analysing Qinghuang power mechanism of therapeutic action.Methods We analyses the changes of DNMT1, DNMT3a and DNMT3b mRNA relative quantitation which measured by a real-time quantitative PCR from30patients before and after treatment with Qinghuang powder. Peripheral blood cells were detected during the same period by hematology analyzer. After the test,the patients were divided into effective and ineffective groups according to clinical curative effects evaluation. The changes of DNMTs were observed respectively in the two groups. To discuss the correlations between DNMTs mRNA levels and changes in peripheral blood before and after the treatment. Maths model is built up to show the relationship of DNMTs mRNA levels and changes in peripheral blood using multiple linear regression analysis of experimental results,which can predict the change of blood cells by DNMTs mRNA content.Results1.30cases of patients were evaluated by clinical curative effects evaluation,23patients were effective,7patients were ineffective, effective rate was76.67%;2Both Hgb and PLT were significantly increased effective group(p<0.05), nevertheless,the ineffective group was not(p>0.05).3.the level of DNMT1mRNA were decreased significantly in23effective cases compared with those before treatment (p<0.05), no significant changes were found in7ineffective cases (p>0.05)4. DNMT1DNMT3a DNMT3b mRNA relative content in valid group were lower than those in invalid group before treatment (P<0.05),The result was similar after treatment (P<0.05).5. The WBC change values demonstrated a negative correlation with DNMT3a mRNA relative quantitative logarithmic before treatment. their correlation coefficient is-0.41, The Hgb change values demonstrated a negative correlation both with DNMT1and with DNMT3a mRNA relative quantitative logarithmic before treatment. The correlation coefficient is-0.57and-0.67.The PLT change values demonstrated a negative correlation with DNMT1,DNMT3a and DNMT3b mRNA relative quantitative logarithmic before treatment.and also with DNMT3a mRNA. Correlation coefficient were-0.47,-0.37,-0.62and-0.41respectively.6.The three DNMTs mRNA factors affecting blood cells were analysed by multiple regression method and optimum equations were suggested for the main factor-Dnmtl mRNA relative quantitative logarithmic before treatment affecting Hgb and PLT.Conclusion Qinghuang power mainly reflected on inhibition of DNMT1mRNA expression, It exert the therapeutic action on improving blood cells level by way of reducing methyltransferase activity and reducing methylation process from SAM to DNA. Part Two To study effect of Qinghuang power treatment on S-adenosylmethionine meta-bolic circulation in MDS patientsObjective To study the effect of Qinghuang power on MDS patients’ S-adenosylmethionine metabolic circulation and observe the correlations between changes in peripheral blood cells and SAM with its metabolic products. The Research Article focus on analysing Qinghuang power methylation pathway.Methods We determined the serum contents of SAM,SAH,HCY measuring by Elisa from54patients before and after treatment with Qinghuang powder. Peripheral blood cells were detected during the same period by hematology analyzer. After the test,the patients were divided into effective and ineffective groups according to clinical curative effects evaluation and chromosomal karyotypes.The changes of SAM,SAH,HCY contents were observed respectively in the two groups. To discuss the correlations between changes of SAM,SAH,HCY contents and changes in peripheral blood before and after the treatment.Results1.54cases of patients were evaluated by clinical curative effects evaluation,35patients were effective,19patients were ineffective, effective rate was64.81%;2Both Hgb and PLT were significantly increased in effective group (p<0.05), nevertheless,the ineffective group was not (p>0.05).3. No significant SAM content changes were observed in effective group (p>0.05).the level of SAH was decreased significantly after treatment (p <0.05). SAM:SAH ratio was increased after treatment (P<0.05);moreover,HCY contents rose in effective group (P<0.05), nevertheless, above data remained unchanged in19ineffective patients(P<0.05).4.54patients were split into three groups according to different chromosomal karyotypes.We compare the contents of SAM,SAH,HCY differences among three groups,and After statistics processing, the experiment result shows no evident differences among them (P>0.05).5. The Hgb change values demonstrated a positive correlation with HCY content before and after treatment. their correlation coefficient is0.33(P<0.05), The PLT change values demonstrated a positive correlation with HCY content before and after treatment.Their correlation coefficient is0.35(P<0.05)Conclusion Qinghuang power influences SAM-SAH-HCY metabolic circulation.The inhibition of DNMT1mRNA induced by Qinghuang power causes metabolism weakness from SAM to SAH,which results reducing Methyl group transferred from SAM to DNA.