| Squid ink melanin is an irregular polymer consisting of an indole structure, which can chelate many kinds of metal ions. Squid ink melanin-Fe is prepared by its compexation properties of melanin. In this study, high-purity squid ink melanin was ertracted from squid ink sac. This paper made a comprehensive assessment of the safety of squid ink melanin-Fe by acute toxicity, genetic toxicity and30-days-feeding test. This paper was to explore ameliorative effect and mechanism of squid ink melanin-Fe on iron deficiency anemia rat model and myelosuppressed mice model. What’s more, the absorptive characteristics of squid ink melanin-Fe was studied using everted intestinal sac method and in vivo test. The main researches in this thesis are listed below:Squid ink melanin-Fe was safe to be taken orally. The result of acute toxicity test showed that, after being given squid ink melanin-Fe, no any other harmful symptoms were observed and no mice were dead. And the maximum tolerable dose(MTD) was larger than18000mg/kg-bw. All the results of Ames test, bone marrow cell micronucleus test and sperm abnormality test in mice were negative. There was no significant genetic toxicity of squid ink melanin-Fe. Squid ink melanin-Fe had no obvious damage to the liver, kidney and cardivascular according to investigating the biochemical index of blood and urine in rats during the30days oral toxication experiment.It appeared to promote the restoration of IDA. Squid ink melanin-Fe also had significant effects on stimulating erythropoiesis in IDA rats. It was evident that squid ink melanin-Fe significantly enhanced the number of red blood cell(RBC), contents of hemoglobin(Hb), hematocrit(HCT), mean corpuscular volume(MCV), serurn iron(SI) and serum erythropoietin(EPO)content, as well as reduced serum total iron binding capacity(TIBC)content and the concentration of free erythrocyte prolopohyrin(FEP) in RBC. The hematopoiesis function was studied by bone marrow cell and colony culture. It was proved that squid ink melanin-Fe increase the number of granulocyte/monocyte colony forming units(CFU-GM) and colony forming unit-erythrocyte(CFU-E). Detected with RT-PCR, it was find out that squid ink melanin-Fe could promote GM-CSF and EPO mRNA expression bone marrow cell. Therefore, the results indicate that squid ink melanin-Fe can stimulate secretion of GM-CSF and EPO to promoting hematopoiesis function of bone marrow in rat.The investigation of the stimulating erythropoiesis mechanisms of squid ink melanin-Fe were conducted by RT-PCR. The results suggested that squid ink melanin-Fe could improve the mRNA expression level of Raf-1, MEK1/2, ERK1/2, c-Jun and c-Fos. It is suggested that squid ink melanin-Fe could stimulate erythropoiesis process involving commitment, proliferation and differentiation by enhancing Raf-1/MEK/ERK signal transduction pathway.The hematopoiesis function was studied in myelosuppressed mice model which was induced by there intraperitoneal injections of Cyclophosphamide. It is suggested that squid ink melanin-Fe could increase the number of CFU-GM and CFU-E, stimulate secretion of GM-CSF and EPO to promoting hematopoiesis function of bone marrow and enhance the number of peripheral blood WBC, RBC and Hb in mice.Rat evered gut sac system and in vivo test were used for the study on absorption characteristics of squid ink melanin-Fe. The results indicated that squid ink melanin-Fe could be uptaked in an intact form by the epithelium absorptive cells. Serum iron concentration would be the maximum level at2hours after intragastric administration of squid ink melanin-Fe.In a summary, all the results of the study showd that squid ink melanin-Fe was safe to be taken orally. Squid ink melanin-Fe could promote the hematopoietic progenitor cells to differentiate along granulocyre-macrophage and erythroid line, stimulate erythropoiesis by enhancing Raf-1/MEK/ERK signal transduction pathway and be uptaked intact. |
PDF Full Text Request