[1]张舟,陈芳,曾东风,等.TMP线性二联体多肽的筛选及其对辐射损伤小鼠血小板减少症的救治作用[J].第三军医大学学报,2013,35(19):2005-2009.
 Zhang Zhou,Chen Fang,Zeng Dongfeng,et al.Screening for dimmer of thrombopoietin mimetic peptide with potent thrombo-cytopoietic effect and its effect on radiation-induced thrombocytopenia in mice[J].J Third Mil Med Univ,2013,35(19):2005-2009.
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TMP线性二联体多肽的筛选及其对辐射损伤小鼠血小板减少症的救治作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第19期
页码:
2005-2009
栏目:
论著
出版日期:
2013-10-15

文章信息/Info

Title:
Screening for dimmer of thrombopoietin mimetic peptide with potent thrombo-cytopoietic effect and its effect on radiation-induced thrombocytopenia in mice
作者:
张舟陈芳曾东风许杨王崧申明强陈默陈石磊粟永萍王军平
第三军医大学军事预防医学院全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室
Author(s):
Zhang Zhou Chen Fang Zeng Dongfeng Xu Yang Wang Song Shen Mingqiang Chen Mo Chen Shilei Su Yongping Wang Junping
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, College of Military Preventive Medicine, Third Military Medical University, Chongqing, 400038, China
关键词:
TMP二联体多肽巨核细胞血小板辐射损伤
Keywords:
dimer of thrombopoietin mimetic peptide megakaryocyte radiation injury platelet
分类号:
R341; R394-33; R558.205
文献标志码:
A
摘要:
目的      筛选具有显著促进巨核细胞增殖、分化和血小板生成活性的血小板生成素模拟肽(TMP)二联体线性多肽,并观察其对血小板减少症的救治作用。      方法       设计合成系列TMP二联体线性多肽,其中两个头尾串联的TMP之间设置不同长度的连接肽,CCK-8法检测并筛选促巨核细胞增殖活性最强的TMP二联体多肽,Western blot分析TMP二联体多肽激活STAT5和ERK1/2磷酸化情况,流式细胞术分析小鼠骨髓来源Sca+-Ⅰ细胞经TMP二联体多肽处理后巨核细胞标志分子CD41、CD61的表达变化。BALB/c小鼠用5 Gy γ射线全身一次性照射后皮下注射TMP二联体多肽,每天1次,连续用药7 d,分析外周血血小板水平变化和骨髓造血组织病理改变。      结果       连接肽长度和氨基酸组成可显著影响TMP二联体多肽的促巨核细胞增殖活性,其中由1个脯氨酸(P)和8个甘氨酸(G8)连接的TMP二联体活性较为突出。TMP-PG8-TMP多肽处理能够显著促进巨核细胞中STAT5和ERK1/2的磷酸化,该多肽连续作用14 d能够较IL-3对照组显著上调Sca+-Ⅰ细胞CD41、CD61的双阳性表达率[(83.26±5.38)% vs (15.70±2.12)%,P<0.01]。此外,体内应用该多肽能够较辐照对照显著提高急性辐射损伤小鼠外周血血小板最低值水平(164.8×109/L vs 53.0×109/L),并使血小板水平提前4 d恢复。      结论       通过筛选连接肽最终获得具有显著促巨核细胞增殖和血小板生成活性的TMP二联体线性多肽。
Abstract:
Objective      To screen the dimmer of thrombopoietin mimetic peptide (dTMP) with potent thrombocytopoietic effect on the promotion of thrombopoiesis and the proliferation and differentiation of megakaryocyte.       Methods      A series of linear peptides containing two TMP joined by peptide linkers at different lengthes were designed and synthesized. The promoting activity of the dTMP on megakaryocyte proliferation was determined by cell counting kit-8 (CCK-8) assay. The ability of dTMP to phosphorylate ERK1/2 and STAT5 signal pathways was measured by Western blotting. The expression level of CD41/CD61 was analyzed by flow cytometry to reveal the ability of dTMP peptide to promote murine marrow Sca+-Ⅰ cells to differentiate into megakaryocytes. Finally, dTMP was administered to BALB/c mice with thrombocytopenia induced by 5 Gy total body irradiation through subcutaneous injection, in order to investigate the effect of dTMP on platelet recovery in vivo.       Results      The length of peptide linkers significantly influenced the promoting activity of dTMP peptides on megakaryocyte proliferation. The dTMP peptide with a linker composed by a proline and eight glycine (TMP-PG8-TMP) displayed a remarkable effect on megakaryocyte proliferation. Western blot assay demonstrated that TMP-PG8-TMP had the ability to activate the phosphorylation of ERK1/2 and STAT5. The expression level of CD41/CD61 in the cells derived from Sca+-Ⅰ marrow cells treated with TMP-PG8-TMP was much higher than that of IL-3 control group [(83.26±5.38)% vs (15.70±2.12)%, P<0.01]. Meanwhile, dTMP-treated mice were about 4 d earlier in the platelet recovery than that of saline control, demonstrating that this peptide was able to be used for the treatment of thrombocytopenia.       Conclusion      A dTMP peptide with potent thrombocytopoietic activity is finally obtained by peptide linker screening.

参考文献/References:

张舟,陈芳,曾东风,等. TMP线性二联体多肽的筛选及其对辐射损伤小鼠血小板减少症的救治作用[J].第三军医大学学报,2013,35(19):2005-2009.

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更新日期/Last Update: 2013-09-30