重组人血小板生成素促进急性放射病小鼠长期造血功能恢复的作用及相关机制研究  

Effect of Recombinant Human Thrombopoietin(rhTPO)on Longterm Hematopoietic Recovery in Mice with Acute Radiation Sickness and Relative Mechanism

作　　者：栾皓 邢爽 杨锦坤 王叶梅 张雪文 乔子肢 申星 余祖胤 LUAN Hao;XING Shuang;YANG Jing-Kun;WANG Ye-Mei;ZHANG Xue-Wen;QIAO Zi-Zhi;SHEN Xing;YU Zu-Yin(School of Life Science,Anhui Medical University,Hefei 230032,Anhui Province,China;Beijing Key Laboratory for Radiobiology,Beijing Institute of Radiation Medicine,Beijing 100850,China;Graduate School of Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China)

机构地区：[1]安徽医科大学生命科学学院,安徽合肥230032 [2]军事科学院军事医学研究院辐射医学研究所,北京100850 [3]锦州医科大学研究生学院,辽宁锦州121001

出　　处：《中国实验血液学杂志》2023年第2期546-552,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(81773366,81573088,81202147);军队后勤科研计划项目(AWS21J010)。

摘　　要：目的:研究重组人血小板生成素(recombinant human thrombopoietin,rhTPO)促进急性放射病小鼠长期造血功能恢复的作用与机制。方法:小鼠经^(60)Coγ射线(6.5 Gy照射)全身照射后2 h肌肉注射rhTPO(100μg/kg),于照射后6个月检测小鼠外周血象、骨髓造血干细胞(HSC)比例、竞争移植受体小鼠存活率及嵌合率、c-kit^(+)HSC衰老阳性率、以及c-kit^(+)HSC衰老相关基因p16、p38 mRNA表达变化。结果:经6.5 Gy(63.01 cGy/min)γ射线照射后6个月,正常对照组、照射对照组和rhTPO给药组小鼠的外周血白细胞、红细胞、血小板、中性粒细胞和骨髓有核细胞数均未见明显差异(P>0.05);照射后小鼠骨髓造血干细胞和多能祖细胞比例较正常对照组显著下降,但rhTPO组无明显改变(P>0.05);照射对照组小鼠CFU-MK和BFU-E的数量显著低于正常对照组,rhTPO给药后明显改善;小鼠骨髓细胞竞争移植后,正常对照组和rhTPO给药组受体小鼠70 d存活率均为100%,照射对照组受体小鼠全部死亡;正常对照、照射对照和rhTPO给药组照射后6个月小鼠c-kit^(+)HSC衰老阳性率分别为6.11%、9.54%、6.01%(P<0.01);照射对照组小鼠的c-kit^(+)HSC p16、p38 mRNA表达显著高于正常对照组(P<0.01),rhTPO给药后降低明显(P<0.01)。结论:6.5 Gyγ射线全身照射后6个月小鼠造血功能仍有所下降,提示存在长期损伤。应用rhTPO大剂量给药对其有明显改善作用,通过p38-p16通路减轻照射后长期HSC衰老,可能是rhTPO改善急性放射病小鼠造血功能长期损伤的机制之一。Objective:To investigate the effect and relative mechanism of Recombinant Human Thrombopoietin(rhTPO)on long-term hematopoietic recovery in mice with acute radiation sickness.Methods:Mice were intramuscularly injected with rhTPO(100μg/kg)2 hours after total body irradiation with ^(60)Coγ-rays(6.5 Gy).Moreover,six months after irradiation,peripheral blood,hematopoietic stem cells(HSC)ratio,competitive transplantation survival rate and chimerization rate,senescence rate of c-kit^(+)HSC,and p16 and p38 mRNA expression of c-kit^(+)HSC were detected.Results:Six months after 6.5 Gyγ-ray irradiation,there were no differences in peripheral blood white blood cells,red blood cells,platelets,neutrophils and bone marrow nucleated cells in normal group,irradiated group and rhTPO group(P>0.05).The proportion of hematopoietic stem cells and multipotent progenitor cells in mice of irradiated group was significantly decreased after irradiation(P<0.05),but there was no significant changes in rhTPO group(P>0.05).The counts of CFU-MK and BFU-E in irradiated group were significantly lower than that in normal group,and rhTPO group was higher than that of the irradiated group(P<0.05).The 70 day survival rate of recipient mice in normal group and rhTPO group was 100%,and all mice died in irradiation group.The senescence positive rates of c-kit^(+)HSC in normal group,irradiation group and rhTPO group were 6.11%,9.54%and 6.01%,respectively(P<0.01).Compared with the normal group,the p16 and p38 mRNA expression of c-kit^(+)HSC in the irradiated mice were significantly increased(P<0.01),and it was markedly decreased after rhTPO administration(P<0.01).Conclusion:The hematopoietic function of mice is still decreased 6 months after 6.5 Gyγ-ray irradiation,suggesting that there may be long-term damage.High-dose administration of rhTPO in the treatment of acute radiation sickness can reduce the senescence of HSC through p38-p16 pathway and improve the long-term damage of hematopoietic function in mice with acute radiation sickness.

关 键 词：重组人血小板生成素 急性放射病 长期骨髓抑制 

分 类 号：R730[医药卫生—肿瘤] R818[医药卫生—临床医学]