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x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(11.82±3.24)%,高于对照组〔(2.07±2.21)%,P<0.05〕、细胞周期G1期阻滞,G0/G1期比例为(86.70±3.79)%,高于对照组〔(69.31±3.07)%,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。
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 x - ray for one - time total body irradiaticn （ TBI }, within 6h , lntraperitoneal injection of phonylhydrasine hydreehloride 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- galactasidase {SA- β- gal）cytochemical staining, The ratio of aging Sca- 1^＋ HSC/HPC entered G1 phase of the cell cycle, spleen index and biopsy. Results：Compared with the control group, apathetic expression, behavioral retardatien,decreased food intake were observed, the routine bloed test were dramatically declined, the number of MNCs and Sca- 1^＋ HSC/HPC were significantly reduced, the positive staining rate and the ratio of Sca - 1^＋ HSC/HPC were higher in model group than in control group （11.82% ± 3.24% vs 2.07%±2.21% ,86.70%±3.79% vs 69.31%±3.07%, P 〈 0.05）, the spleen index was significantly reduced and the spleen had Significant degenerative changes. Conclusion： The way of usingx- ray and phenylhydrazine hydrochloride can successfully establish the model of deficiency qi - blood and induce Sca- 1 ^＋ HSC/HPC aging.
出处
《激光杂志》
CAS
CSCD
北大核心
2012年第4期88-89,共2页
Laser Journal
基金
国家自然科学基金面上项目(81173398)
