| Aplastic anemia (aplastic anemia, AA) is by a variety of causes, various pathogenesis causes medullary hematopoiesis function exhaustion, caused by the decrease in the number of hematopoietic cells, to anemia, bleeding and infection as the main symptoms of a group of heterogeneous disease. Now think its incidence main proliferation with hematopoietic stem cell inner defects, hematopoietic microenvironment function defects and the body's immune dysfunction related. Hematopoietic microenvironment mainly by the bone marrow stromal cells constitutes, past the study of hematopoietic microenvironment spend on the basis of matrix components. Along with the experiment technology unceasing enhancement, the people have to have been isolated from bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells, BMSC). The origin of mesoderm BMSC are primitive cells, is marrow stromal cell lines of stem cell precursors of bone marrow microenvironment, is an important part of this research is to study, patients with normal BMSC AA cells form, cells BMSC cycle, the growth curve, immunophenotype, differentiation potential and secretory stem cell factors (SCF) level of biological characteristics of differences, thus further research GATA-1, GATA-2, PPAR gamma and SCF, AA patients and normal genes in the difference, expressed in BMSC and presented according to the experimental results, a new method of treatment, AA clinical application BMSC treatment to provide the evidence that aplastic anemia. The first partObjective:to study the biological characteristics of AA patients BMSC changes.Methods:the normal bone marrow for comparison, observe the BMSC cell type, growth curve, streaming cells cell cycle, the detection instrument immunophenotype, ELISA test AA patients with normal BMSC secretion BMSC SCF distinction, take the above-mentioned income source of amplification cells monoclonal separately to fat cells and the osteoblast directional induction.Results:the AA patients BMSC for stick wall fibroblasts kind, the original generation cells form uneven, third generation cells after covered with bottle a long spindle, swirling distribution uniformity, AA group, form time longer in growth. Compared with normal AA patients BMSC surface of BMSC, the results show that both sign no obvious difference, all don't express CD34 expression CD106 CD29, and CD44. ELISA test and AA patients SCF level, AA group (95.6±1.45) pg/ml and the group of BMSC(125.28+18.12) pg/ml, (P= 0.002, P< 0.01), has a statistically significant difference.Conclusion:through our AA patients to the biological characteristics of BMSC preliminary research, AA BMSC phenotype and known, and maintain normal BMSC consistent multiplex differentiation potential, but its growth slow growth, exist obvious defects and quality defects, number may be contributing to the marrow microenvironment AA one of the reasons the defects.The second partPurpose:to test GATA-1, GATA-2, and SCF, gamma PPAR gene in the AA patients and normal BMSC expression.Methods:using fluorescence quantitative polymerase chain reaction (PCR) detection GATA-1, GATA-2, SCF PPAR gamma, and gene in the AA patients and normal BMSC expression. Results:compared with normal BMSC patients, AA GATA-2 and the BMSC SCF gene expression and decreased (Relative express quantity are 0.36±0.08,0.42±0.21); But PPAR gamma gene expression quantity is elevatory (Relative express quantity 1.51±0.195); Compared with control GATA-1, no significant difference (Relative express quantity 0.91±0.06)Conclusion:GATA-2, SCF differentially expressed genes may affect the microenvironment AA patients bone marrow hematopoietic regulation function; Gamma PPAR the expression in AA patients BMSC PPAR gamma, rose significantly differentially expressed genes may explain AA patients into fat bone marrow easy reasons, these genes whether to AA pathogenesis and treatment to provide a new thinking and deserves further investigation.The third partObjective:to determine fat cells induced group and Chinese medicine dispelling the fat meat group two groups PPAR gamma mRNA and within BMSC expression of protein levelMethods:using PT-PCR and Western blotting spectrometry fat cells induced group and Chinese medicine dispelling the fat meat group of two groups of BMSC PPAR gamma mRNA and within the protein level expressionResults:compared with fat cells induced group, Chinese traditional medicine dispelling the fat meat group fat cells has significantly reduced, rate of statistical significance (P= 0.047, P< 0.01), Chinese traditional medicine dispelling the fat meat group mRNA level and PPAR gamma protein level obviously decreased.Conclusion:Chinese medicine dispelling the fat meat to AA BMSC adipose differentiation has inhibition, by reducing fat cells forming, lower fat cells specific PPAR gamma transcription and symbol of protein expression...
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