The Levels Of TPO,TGF-β₁ And TGF-β RⅢ In Bone Marrow In Children With Idiopathic Thrombocytopenic Purpura

Objective Idiopathic thrombocytopenic purpura (ITP) is divided into acute ITP (AITP) and chronic ITP (CITP) in clinic. The standard to divide it is if the course of disease exceed six months. But we considered that the mechanisms and clinical expressions of AITP and CITP were "very complicated. There might be other difference between them. It was identified that effects of cytokines/chemokines on megakaryocytes may take great parts in the happening of ITP. They can affect the proliferation, differentiation and maturity of megakaryocytes. We think that they possibly act differently in the process of AITP and CITP. The main aim of this experiment is to evaluate the levels of positive cytokine thrombopoietin(TPO), negative cytokine transforming growth factor- 1 (TGF- 1) in bone marrow and expression levels of transforming growth factor- 1 receptor IIImRNA (TGF- RIII mRNA) on megakaryocytes in children with AITP or CITP. They can help us to elucidate further the mechanisms of AITP and CITP.Methods (l)Bone marrow were respectively aspirated from 28 children with AITP and 16 children with CITP before treatment. 20 controls were aspirated from children who accepted examination of bone marrow and the results were negative.(2)Put 0.8ml bone marrow into 0.2ml 1%EDTA, 2000r/min centrifugalizing to get the upper liquid and preserve it in -20 癈 refrigerator. (3)Purification of megakaryocytes from bone marrow: Aspirate 1.5ml~3ml bone marrow and put it into modified PBS rapidly, use immunomagnetic beads to isolate megakaryocytes from it. Smear them on a piece of glass slice and preserve it in -20 癈 refrigerator. (4)Measurement of TPCK TGF- P ! and TGF- P RlIImRNA: Use ABC-ELISA to evaluate the levels of TPCK TGF- 3 i in bone marrow and xspZYMED sjtu hybridization to evaluate the expression of TGF- P RlIImRNA on megakaryocytes. (S)Statistical assay: the data were dealt with SPSS 10.0 Statistical software by analysis of variance and linear correlation.Results: l.The levels of TPO in bone marrow: the level of TPO in AITP was 299.72+ 136.30pg ?ml'1, in CITP 391.79 ?104.57 pg ?ml'1 and in controls 273.44 ?112.95 pg ?ml"1. There was no significant difference between AITP and controls (P=0.565). There was significant difference between CITP and controls (P=0.022). There was significant difference between AITP and CITP (P=0.049); 2.The levels of TGF-P 1 in bone marrow: the level of TGF-P l in AITP was 263.39 + 143.88 pg ml-1, in CITP 363.24?136.56 pg ml"1 and in controls 157.74?2.37 pg ml-1. There was significant difference between AITPand controls (P=0.030). There was significant difference between CITP and controls(P=0.000). There was also significant difference between AITP and CITP (P=0.039); 3. TGF- P RlIImRNA positive cell ratio : TGF- P RlIImRNA positive cell ratio in AITP was 84.23 + 8.49%, in CITP 91.33 + 7.93% and in controls 74.80 + 4.77%. There was significant difference between AITP and controls (P=0.000). There was significant difference between CITP and controls (P=0.000). There was also significant difference between AITP and CITP (P=0.006); 4. TGF- P R HI mRNA reactive intensity on megakaryocytes: TGF- P RlIImRNA reactive intensity on megakaryocytes in AITP was 170.00+32.72, in CITP 192.67 + 38.66 and in controls 134.53?18.19. There was significant difference between AITP and controls (P=0.001). There was significant difference between CITP and controls (P=0.000). There was also significant difference between AITP and CITP (P=0.036) ; 5.Correlativity assaybetween TPO and TGF- β 1 in bone marrow: In AITP and controls, there was no exact correlationship between TPO and TGF- β1, the results were respectively r=-0.7 (P>0.05) and r=0.26 (P>0.05). But there was positive correlationship between TPO and TGF- β1j in CITP r=0.653 (P<0.05).Conclusions (1) In AITP, the reason which caused decreasing of platelets was highly destroying of platelets. Effect of this process on TPO was little; in CITP, maturity disturbance of megakaryoctes can cause decreasing of platelets. This process can affect the leve...