中医气虚血虚模型-第三军医大学学报.DOC

中医气虚血虚模型与造血干/祖细胞衰老 刘典锋,周玥,蔡世忠,耿珊，张琛，孙可，王亚平* （400016 重庆,重庆医科大学组织学与胚胎学教研室,干细胞与组织工程研究室） 【摘要】 目的 构建中医“气虚血虚”模型，探讨造血干/祖细胞（HSC/HPC）衰老及检测相关生物学指标,为阐释中医“气虚血虚”的现代生物学机理和探讨“补气生血”药物的作用提供理论与实验依据。方法 6.5Gy的χ射线一次性全身辐射C57小鼠,辐射6h内腹腔注射盐酸苯肼40mg/kg，建立“气虚血虚”模型（即骨髓抑制与贫血模型），分别在第3、7、14、28d观察小鼠的行为学变化、外周血象、骨髓单个核细胞总数、免疫磁性分选法分离鉴定小鼠Sca-1+ HSC/HPC、检测骨髓单个核细胞数（MNCs）与造血干/祖细胞（Sca-1+ HSC/HPC）数量、衰老SA-β-半乳糖苷酶染色检测Sca-1+ HSC/HPC阳性率、Sca-1+ HSC/HPC细胞周期分布、脾指数与脾脏病理变化。结果 与对照组相比，在各时间段上模型组小鼠均表现出精神萎靡，行为迟缓，进食量下降等中医“气虚血虚”症候、外周血象各项指标降低，MNCs数下降，Sca-1+ HSC/HPC数下降、SA-β-半乳糖苷酶染色阳性百分率（11.823.24)%，高于对照组[（2.072.21）%,P＜0.05]、细胞周期G1期阻滞，G0/G1期比例为（86.703.79）%，高于对照组[（69.313.07）%，P＜0.05]，S期比例减少、脾指数降低和脾脏退行性变化显著。结论 辐射和盐酸苯肼并用法可成功的建立中医“气虚血虚”动物模型，并导致造血干/祖细胞衰老。 【关键词】 气虚血虚；动物模型；造血干/祖细胞；衰老 [中文图书分类号]R332;R223.14;R329.28;R339.38 [文献标志码]A Establishment of the model of deficiency qi-blood and aging HSC/HPC LIU Dianfeng,Zhou Yue,Cai Shizhong,Geng Shan,Zhang Chen,Sun Ke,WANG Yaping Department of Histology and Embryology ,Laboratory of Stem Cell and Tissue Engineering，Chongqing Medical University，Chongqing 400016 China) [Abstract] Objective To establish the model of deficiency qi-blood and investigate the aging Sca-1+ HSC/HPC relevant biological indicators. The purpose is to explain the mechanism of deficiency qi- blood and build the foundation of studying the mechanism of qi- blood-tonic. Methods Mice (C57BL/6) were dealt with 6.5Gy χ-ray for one-time total body irradiation (TBI), within 6h , Intraperitoneal injection [基金项目] 国家自然科学基金资助项目 [通讯作者] 王亚平，E-mail: HYPERLINK mailto:ypwangcq@yahoo.cn ypwangcq@yahoo.cn of phenylhydrazine hydrochloride for 40mg/kg.establishing the model of bone marrow suppression and hemolytic anemia. watch the behavior of mice and assay the number of MNCs and Sca-1+ HSC/HPC, senescence-associated β-galactosidase (SA-β-gal) cytochemical staining, The ratio of aging Sca-1+ HSC/HPC entered G1 phase of the cell cycle, spleen index and biopsy. Result