Researching Of The Gene Profiles And The Proteom In Dicidua From Spontaneous Abortion During Early Pregnancy

Decidual tissue is a very complex and highly concordant tiny environment between mother and embryo during pregnancy, which is constituted of the variety cells such as extravi11ous trophoblast(EVT),endometrium cells, bone marrow cells, decidual microphage,T cells and a few B cells and so on. These cells and cell factors excreting from the cells, mother and embryo to the deciduas form information communication platform between mother and embryo, and regulate subtly to moderate invasion of trophblast cells, endometrial immunization compatibility, decidualization and placental formation. If this regulation is overbalanced, the regulation of decidua tiny environment will be breaked. It result in reject reaction of mother, and induce abortion.Early spontaneous abortion is a kind of abortion which end un-intended pregnancy before 12 weeks. The frequence possess about 62% of whole spontaneous abortions in clinics. A part of spontaneous abortion agt. is unclear. In the recent years, with the research of the endometrium reception and the decidual histopathology, it shows that form of the endometrium reception to the embryoblast is the essential for the normal embryo development, and related to normal decidualization. But to the early spontaneous abortion decidual tissue, what happened in its microenviroment? and how about did the internal relationship between the change and the spontaneous abortion? In the past, this searching involved in special gene or special protein, but seldomly involved in the gene expression profile and proteomics. Recently large-scale gene expression profile analysis and proteomics analysis has been used in the research area of biology and medicine widely. Now oligonucleotide array,DNA mircoarray and gene expression serial analysis were widely used to analysis of the gene expression. Human genome U133plus2.0 of the oligonucleotide array contains 39000 transcript, represents 33000 genes known, and consists of two arraies A and B, includes more than millions probes, the sequence are from GenBank,dbEST, refseq data bank. By U133plus2.0 array, we can observe the change between normal decidual tissue and spontaneous abortion decidual tissue. The two-dimensional gel electrophoresis and mass spectrometry may be used to researching the protein expression level and amino acid sequence, protein modifying way after gene translation . For this reason, we have analysized the gene expression profile and protein expression profile in the spontaneous abortion decidual tissue, and acquired the data about expression changes of gene and protein. It provides the important base for etiological diagnosis, precaution and treatment of the spontaneous abortion.By the analysis of the Human genome array UI133plus2.0, we have observed 1876 genes expressed diffferently, the up-regulation 1004, the down-regulation 872. Through searching the biological function of the 1876 differential expression genes, we find that there are 37 up-regulation and 32 down regulation genes related extracellular matrix; 83 up-regulation and 47 down regulation genes related signal transduction; 71 up-regulation genes and 59 down-regulation genes related transcription; 85 up-regulation and 88 down regulation genes related cell growth/maintenance; 103 up-regulation and 86 down-regulation genes related metabolism; 55 up-regulation and 27 down-regulation genes related apoptosis; 67 up-regulation and 77 down-regulation genes related material conveying; 21 up-regulation and 23 down-regulation genes related cell cycle; 9 up-regulation and 3 down regulation genes related alarm reaction; 28 up-regulation and 7 down regulation genes related cytophylaxis; 41 up-regulation and 25 down-regulation genes related immune response; 31 up-regulation and 47 down regulation genes related cell adhesion; 81 up-regulation and 85 down regulation genes related growth. Besides, we find 558 genes have not been described in functions, and the up-regulation 293,down regulation 265 among them. Pathway analysis to the 1318 genes known its function, we find that they mainly refer to the signal pathways of MAPK,ERK, P38, Jak-STAT,Natural killer cell mediated cytotoxicity,mTOR,Apoptosis etc. By bioinformatics analysis to the genes, we think that the spontaneous abortion etiology refers to up- down-regulation of multiple genes, such as genes related apoptosis,immune response,hydrolyase, cell adhesion and so on. These factors may be the main cause which results in spontaneous abortion.2-DE Gel analysis shows protein molecular weight concentrated mainly on 14-116KDa, iso electric point was mainly in range of pH4.0-8.0 in the spontaneous abortion decidua. Analysis by PDQuest software, there are 583 and 613 protein points in spontaneous abortion decidua and normal decidua respectivly. As the standard with the normal decidua, proteins matching rate is 78% between spontaneous abortion decidua and the normal decidua. Protein points of the differential expression are 73(up-regulation 38, down-regulation 35). up-regulation Protein points over three times are 23, and down-regulation Protein points over three times are 17. Proteins only expressed in the spontaneous abortion decidua are 22 , but 17 in the normal decidua. Indentifying 15 differential proteins by MALDI-TOF-MS, 9 proteins are significant(p<0.05). They are Vimentin, Calreticulin precursor, Annexin A5, tropomyosin alpha-4, endoplasmin precursor, protein disulfide-isomerase A3 precursor , Annexin A1,α-enolase. These proteins as Vimentin, Annexin A5, Annexin A1,α-enolase express increasely, and these proteins as Tropomyosin alpha-4,Grp94,Protein disulfide-isomerase A3 precursor express decreasely. Calreticulin precursor only appears in the spontaneous abortion decidua. These differential expression proteins mainly refer to invasion of the blastocyst, blood vessel remodeling and cell apoptosis.We have researched the gene MMP10 expressed highly in spontaneous abortion. This gene expressed mainly in the decidual matrix cells, but in the normal decidua expressed lowly. Gene MMP10 was not observed in endochylema of the abortion decidua which the cell nuclei has condensed and schizolysis. By the experiment of transwll, we have observed MMP10 still can promote trophocyte invasing. From the feature of MMP10, we suggested that high expression of MMP10 might be compensated in the spontaneous abortion. In the process of invasing trophoblast to the decidua in normal pregnancy, the hydrolization of extracellular matrix mainly depends on MMP2 and MMP9,but MMP10 is not essential for the process. From high expression of MMP10 in our the experiment, we suggested that the change of MMP10 expression in the spontaneous abortion decidua might profit for degradation and remodeling of the decidual extracellular matrix, and provides suitable space for the trophoblast invasion and prevent occurrence of abortion, but its precise molecule mechanism still study further.