Application of EPO (erythropoietin) analogue in preparation of medicine for treating sepsis
A technique for sepsis and analogues, which is applied in the field of application of EPO analogues in the preparation of drugs for the treatment of sepsis, to achieve the effect of improving survival rate, reducing mortality rate and promoting expression
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Embodiment 1
[0032] Effect of EPO polypeptide on promoting endotoxin-resistant macrophage production
[0033] 1. Animals:
[0034] C57 / BL6 mice, male, 10-12 weeks old, weighing 20-25 grams, were purchased from the Experimental Animal Center of the Army Military Medical University, and were normally raised in an SPF animal room.
[0035] 2. Cell acquisition and culture:
[0036]Bone marrow cells were obtained from femurs and tibias of wild-type C57BL / 6J mice aged 10-12 weeks, centrifuged and cultured in DMEM medium for 24 hours to collect non-adherent cells. After culturing in DMEM medium containing macrophage colony-stimulating factor (MCSF) for 4 days, add fresh DMEM medium containing MCSF to continue culturing for 3 days, and collect adherent cells as bone marrow-derived macrophages (BMDM). Flow cytometry The technology detects the expression of CD11b and F4 / 80, and the purity can reach more than 98%.
[0037] 3. Experimental method
[0038] BMDM was given LPS (100ng / ml) combined wit...
Embodiment 2
[0043] EPO peptide treatment inhibits gene expression of pro-inflammatory cytokines in septic mice
[0044] 1. Animals:
[0045] C57 / BL6 mice, male, 10-12 weeks old, weighing 20-25 grams, were purchased from the Experimental Animal Center of the Army Military Medical University, and were normally raised in an SPF animal room.
[0046] 2. Experimental method:
[0047] Wild-type C57BL / 6J mice aged 10-12 weeks were given intraperitoneal injection of PBS or EPO polypeptide (100 μmol / kg) combined with LPS (1 mg / kg). After 24 hours, the mice received a second LPS (10 mg / kg). / kg) intraperitoneally. Six hours after the second LPS injection, the peritoneal cavity of the mice was washed with normal saline, and centrifuged to collect the inflammatory cells in the peritoneal cavity; in addition, the mouse spleen, lung, liver, kidney and other tissues were taken, and the pro-inflammatory factor TNF in the above samples was detected by fluorescent quantitative PCR method -α, IL6, IL-1β ...
Embodiment 3
[0052] EPO peptide treatment promotes gene expression of antibacterial and tissue repair molecules in septic mice
[0053] 1. Animals:
[0054] C57 / BL6 mice, male, 10-12 weeks old, weighing 20-25 grams, were purchased from the Experimental Animal Center of the Army Military Medical University, and were normally raised in an SPF animal room.
[0055] 2. Experimental method:
[0056] Get 10-12 weeks old wild-type C57BL / 6J mice, use PBS or EPO polypeptide (100μmol / kg) combined with LPS (1mg / kg) to give mice intraperitoneal injection, 24 hours later, the mice received a second LPS (10mg / kg) kg) intraperitoneally. Six hours after the second LPS injection, the peritoneal cavity of the mice was washed with normal saline, and the inflammatory cells in the peritoneal cavity were collected by centrifugation; in addition, the mouse spleen, lung, liver, kidney and other tissues were taken, and the antibacterial genes MARCO and CNLP, and gene expression differences of the damage repair ...
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