Therapeutic Effect Of Black Chicken Peptide On Mice With Regenerative Anemia And Its Mechanism

Anemia is a common clinical symptom,which is mainly characterized by the decrease of red blood cells and hemoglobin.Anemia can be divided into red blood cell production reduction anemia(such as aplastic anemia),hemolytic anemia,hemorrhagic anemia.Among them,aplastic anemia is caused by a variety of causes of bone marrow hematopoietic stem cells,hematopoietic microenvironment damage and immune function changes,resulting in bone marrow hematopoietic failure syndrome,the incidence rate is 7.4 / 100000.At present,the common treatment is the combination of glucocorticoid and androgen.This method has a certain curative effect,but it will produce side effects,such as liver function damage,renal function changes and so on.Therefore,the research and development of functional food for anemia has important practical significance.The resources of traditional Chinese medicine in China are rich in ingredients for the treatment of anemia.As a traditional Chinese medicine,the function of black chicken in treating anemia has long been reflected in Wuji Baifeng pills.To explore the function of black chicken in the treatment of anemia and to prepare anti-anemia active peptides by enzymatic method is of great significance in revealing the anti-anemia mechanism of traditional Chinese medicine black chicken,improving the anti-anemia effect of black chicken and developing adjuvant therapy products without toxic and side effects.Objective:By hydrolyzing black chicken white meat into polypeptide,the therapeutic effect of black chicken meat on mouse anemia model was detected;its mechanism was verified to be immune regulation and erythropoietin expression;to determine the regulatory effect of black chicken peptide on intestinal flora,can further regulate immunity,so as to improve the function of anti-anemia.Methods:1.The preparation of black chicken peptide is to grind fresh black bone chicken into meat paste,which is hydrolyzed by chymotrypsin(the amount of enzyme is 1%).2.24 male Kunming mice were randomly divided into four groups: normal control group(NC group,n = 6),5-fluorouracil group(5FU group,n = 6),black chicken peptide treatment group(5FU+CP group,n = 6),and black chicken homogenate treatment group(5FU+CH group,n = 6).NC group was fed normally without any treatment.5FU group,5FU+CP group and 5FU+CH group were intraperitoneally injected with 150mg/kg 5-fluorouracil,5FU group was perfused with normal saline of the same volume as 5FU+CP group,5FU+CP group was perfused with black chicken peptide of 100mg/kg,100mg/kg chicken homogenate was given by stomach for 10 days,and their body weight,red blood cell count and hemoglobin were monitored.On the10 th day,the mice were killed under aseptic condition,and the serum,colon,kidney and other tissues were taken for pathological and immunological detection.3.The molecular weight of silky chicken peptide was evaluated by SDS-PAGE(polyacrylamide gel electrophoresis),the anemia of mice in each group was evaluated by red blood cell count and ELISA,the distribution of intestinal bacteria in mice was analyzed by PCR-DGGE(denaturing gradient gel electrophoresis),and the changes of intestinal mechanical barrier in mice were judged by Hallie staining and immunohistochemistry.The changes of erythropoietin and immune cytokines in mice in each group were detected by RT-PCR and ELISA techniques,and the mechanism of black chicken peptide in restoring hematopoiesis of anemic mice was analyzed.Results:1.A small molecular peptide was obtained by SDS-PAGE.2.Through the detection of red blood cells and hemoglobin of mice in each group,it was found that 5-fluorouracil inhibited the hematopoietic function of mice to a certain extent,and the hematopoietic function of mice was improved after oral administration of black chicken peptide.and the effect of black chicken homogenate group was not obvious.3.After oral administration of black chicken peptide,the expressions of erythropoietin(EPO),interleukin 3(IL-3)and interferon gamma(IFN-γ)in serum and kidney of mice were increased.3.PCR-DGGE results showed that the intestinal flora of the model group was significantly different from that of the normal group,and the recovery group improved this situation to a certain extent,and the intestinal barrier of mice was damaged after injection of 5-fluorouracil,but recovered to some extent after intragastric administration of 5-fluorouracil.Conclusion:1.Black chicken peptide has a restorative effect on the model of 5-fluorouracil anemia,and the mechanism is related to the promotion of the expression of erythropoietin and the regulation of the expression of interleukin-3 and interferon-γ.2.Black chicken peptide may indirectly promote the balance of intestinal flora by regulating the immune system,and then further promote the recovery of hematopoietic function in mice through the gut-kidney axis.