The Differential Proteomics Study On Blood Enrichment Effects Of Angelica Decoction On Blood Deficiency Mice Model

In this study, the blood deficiency model mice were established by injecting N-acetylphenylhydrazine and cyclophosphamide (ip). Angelica sinensis decoction was given respectively by gastric perfusion by different dose groups (High-dose:20g/kg, Middle dose: lOg/kg, Low-dose:5g/kg). The blood indexes of RBC, WBC, PLT and HGB were detected by hematology analyzer and organ indexes of spleen and thymus were calculated. The blood bi-ochemical parameters of TP, ALT, AST and LDH in the serum were detected by biochemical analysis. In the meantime, the different groups of differentially expressed proteins were ana-lyzed by using two-dimensional electrophoresis (2-DE) and proteomics technology. Combin-ing the above index changes illustrated the mechanism of angelica blood tonic effect. The re-sults were as follws:1. Among three kinds of ways to build blood-deficient model including Hydracetin me-thod, Cyclophosphamide method and both,we found that under the way of model established by Hydracetin and Cyclophosphamide, RBC, WBC, PLT and HGB content in mice of the model group decreased significantly, and mice would appear some blood deficiency symptoms including malaise,skin pale and so on. And using this method to establish model, animals would live long and the blood deficiency state would last long. So APH and CTX coalition building method was a better way.2. Under the model established by APH and CTX coalition building method, the influ-ence on blood deficiency model of different doses of angelica decoction was studied. Com-pared with normal group, mice in model group appear much blood deficiency symptoms such as weak, pale skin and liver and spleen obviously enlarged, the index of spleen and thymus, RBC and HGB content decreased significantly.the content of Na+K+-ATP, Ca2+Mg2+-ATP,ALT,TP,AST and LDH in the serum decreased obviously in the model group. Compared with model group, the RBC, WBC, Hb, Pit and index of thymus gland spleen of the mice were superior to the control group in the Angelica different dose groups. The Angelica different dose groups all had different degrees of increase in the coentent of Na+K+(?) Ca2+Mg2+-ATP, ALT, TP, AST and LDH in the serum.But among the different doses ( lica decoction, the effect of the middle dose(10g/kg) was the best.3. Though Mass spectrometry analysis the11differentially expressed protein spots were detected (Reference standard for protein expression differences more than1.5times, P<0.05),9protein points had Fibrinogen beta chain precursor, Heat shock protein90, Catalase, Glyce-raldehyde-3-phosphate dehydrogenase,Carboxylesterase,Arginase-1,Carbonic anhydrase, S-adenosylmethionine synthase isoform type-1, Heme-binding protein, and two unknown protein were successfully identified by MALDI-TOF-MS. Compared with blank group,9pro-tein spots had changed significantly in model group; The Rise of Heat shock protein90and the fall of Catalase, Glyceraldehyde-3-phosphate dehydrogenase,Carboxylesterase, Arginase-1, Carbonic anhydrase, S-adenosylmethionine synthase isoform type-1, Heme-binding protein. Compared with model group, the above all kinds of protein was reversed to some extent in medicated group (p<0.05). Carbonic anhydrase, Glyceraldehyde-3-phosphate dehydrogenase and Heme-binding protein main effect at RBC participated in energy metabolism and hema-topoiesis of animals’life activities. Carbonic anhydrase, Glyceraldehyde-3-phosphate dehy-drogenase and Heme-binding protein expressed to rise in medicated group, this showed that Angelica sinensis could enhance Carbonic anhydrase, Glyceraldehyde-3-phosphate dehydro-genase and Heme-binding protein content, promote tissue cells involved in hematopoiesis and give play to the role of blood. Heat shock protein90was related to tissue cell differentiation and apoptosis. The content of Heat shock protein90droped in medicated group, this showed that Angelica sinensis could inhibit the apoptosis of animal tissue cells and reduce tissue in-jury.In conclusion, mechanism of Angelica sinensis blood tonic may be relate to the increase of Carbonic anhydrase, Glyceraldehyde-3-phosphate dehydrogenase and Heme-binding pro-tein, which could promote the hematopoietic function of tissue cells and reduce the Heat shock protein90, which could restrain organization cell apoptosis and increase immune organs, promote the body life activity.