The Research On Differences In Gene Transcriptome Profile Of Kidney-yang Deficiency With Ovulation Barrier Sterility

Objective:Choice Kidney-yang Deficiency with Ovulation barrier sterility patients as the object of research, product gene expression profiling of kidney-yang deficiency syndrome, screening of differential gene transcriptional profiles to discuss the molecular pathways and molecular mechanisms of kidney-yang deficiency syndrome, the study will provide a scientific basis on"kidney dominating reproduction".Method:1、Case selection:Yang deficiency constitution and Kidney-yang Deficiency with Ovulation barrier sterility patients were select at the Second Affiliated Hospital of Chengdu University of Traditional Chinese Medicine in gynecology of traditional Chinese medicine clinic From2011June to2011December, Choose the typical kidney-yang deficiency patients as the case group and the typical flat and healthy persons as control group.2、Sample making of Gene chip:Extract peripheral blood of each object, Serial number and then isolate white cells, place the1.5ml EP tube with leukocytes in the negative liquid nitrogen quickly to be preserved, Timely embed the packaged samples in Dry ice and send them to Shanghai Biotechnology Corporation by delivery company.3、The experiments of Gene microarray:Inspected quality of the samples, Tt can enter the stage of experiment after passing all. Extracted and purifiered RNA to complete the probe preparation, then hybrided and scanned the chip, used of gene chip technology for hybridization and scanning analysis. Then the data of expressed genes between patients group and control group are obtained. At last, used SAS statistical software to complete the screening of differences in gene expression.(P<0.05)4、Combination of SAS software, Use GO functional enrichment and KEGG pathway to analysis differential gene expression.Result:1、Totally10cases were enrolled with4cases rejection, and divided into the patients group and control group with3and3cases separately.2、There is characteristic profiles of differential expressed genes between typical kidney-yang deficiency group and normal control group, Among a total of650differential expressed genes, Increase the expression level has266, Reduced expression level there is the384.The Reduced expression level gene has Account for 59.1%proportion.There are480differential expressed genes that obtain gene names and basic information.3、Through the GO functional enrichment analysis, The differential expressed genes are predominantly involved in several biological processes such as structural constituent of ribosome、ribonucleoprotein complexvesicle targeting、establishment of protein localization、chromosome segregation、tissue remodeling、cell leading edge、 positive regulation of locomotion、positive regulation of locomotion.(P<0.05)4、Through the KEGG pathway analysis, These differentially expressed genes involves a number of signal transduction pathway, such as, Metabolic pathways、 TGF-beta signaling pathway、genetic information processing pathways such as DNA splicing and oocyte meiosis、metal ion pathways, intracellular pathways such as immunity, and tumor necrosis factor signaling pathways etc.(P<0.01).In particular, metabolic pathways and TGF-beta signaling pathway is the most outstanding.5、In this study, we found gene associated with germline:PRSS21、TP53TG3、 LAST1、CCR4、SEC24A。Conclusions:1、Gene-chip is a good development prospect method of studying TCM Syndrome associated genes.2、The method of Combining of Ovulation barrier sterility disease and Kidney-yang Deficiency syndrome has enriched the gene pool and deepened the microcosmic mechanism of molecular biology of Kidney-yang Deficiency syndrome.At the same time, the study provides objective basis on"Kidney governing reproduction".3、Ribosomal protein ubiquitination and metabolic dysfunction are closed related with Kidney-yang Deficiency with Ovulation barrier sterility.