黄芪和当归配伍对小鼠造血干细胞衰老模型细胞增殖的影响  

Effects of Astragali Radix combined with Angelicae Sinensis Radix on the proliferation of hematopoietic stem cells senescence model in mice

作　　者：张珂胜 陈凌波[1] 黄小平[1] 邓常清[1] 

机构地区：[1]湖南中医药大学中西医结合心脑疾病防治湖南省重点实验室,湖南长沙410208

出　　处：《中国中药杂志》2017年第21期4187-4194,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81473581);湖南省高校科技创新团队项目;湖南省自然科学创新群体基金

摘　　要：研究黄芪和当归配伍对小鼠造血干细胞衰老模型细胞增殖能力的影响及其作用机制。大鼠灌胃给药制备含药血浆后,体外培养小鼠造血干细胞,实验设空白对照组、模型组、空白血浆组、芪归1∶1血浆组、芪归10∶1血浆组、单用当归血浆组、单用黄芪血浆组,以三丁基过氧化氢(t-BHP)诱导小鼠造血干细胞衰老,以含药血浆作用于造血干细胞衰老模型。SA-β-半乳糖苷酶染色法检测细胞衰老率,流式细胞术检测细胞周期分布,RT-PCR检测Cyclin D1,P21,P53 mRNA表达,Western blot检测Cyclin D1蛋白表达。结果显示,t-BHP作用于造血干细胞后,可使衰老细胞增加,细胞增殖能力降低,G0/G1期细胞增多,G2/M+S期细胞减少,同时伴Cyclin D1表达降低,P53,P21表达增强。黄芪、当归、芪归1∶1配伍、芪归10∶1配伍含药血浆可降低衰老细胞阳性率,使G0/G1期细胞减少,G2/M+S期细胞增加,细胞增殖能力增强,并使Cyclin D1基因和蛋白表达增强,P53,P21基因表达降低。以上效应以芪归1∶1配伍的作用为强。结果表明,t-BHP可使造血干细胞衰老,细胞增殖能力降低。黄芪、当归及其配伍可抑制造血干细胞衰老,促进造血干细胞增殖和细胞周期转换,以芪归1∶1配伍的作用为强。其作用机制可能与其上调细胞周期正性调节因子表达,下调细胞周期负性调节因子表达,从而促进细胞由静止期进入增殖期有关。The aim is to study the effect and its mechanism of Astragalus Radix combined with Angelicae Sinensis Radix on the pro- liferation of hematopnietic stem cells （HSCs） in senescence model. After drug-containing plasma of rats was prepared via intragastric administration, HSCs of mice were cultured in vitro, and then they were divided into blank control group, model group, blank plasma group, Astragalus Radix ＋ Angelieae Sinensis Radix 1：1 group and ｜0：1 group, Angelicae Sinensis Radix plasma group, and Astraga- ｜us Radix plasma group. HSCs senescence model was induced by using tert-butyl hydrogen peroxide（ t-BHP）, and intervened by drug- containing plasma. Cells senescence rate was tested by SA^-galactosidase staining method; cell cycle distribution was determined by flow cytometry; Cyclin D1, P21, and P53 mRNA were measured with RT-PCR, and Cyclin D1 protein expression was measured by Western blot. Results showed that after being induced by t-BHP, senescence rate of HSCs was increased; cell proliferation ability was decreased; count of G0/G1 phase ceils was increased; count of G2/M ＋ S phase cells was reduced; Cyclin D1 expression was down-reg- ulated while P53, P21 expression was up-regulated, which were reversed by Astragalus Radix ＋ Angelicae Sinensis Radix 1： I and 10： 1, single Angelicae Sinensis Radix, and single Astragalus Radix plasma. Furthermore, the above effects were most obvious in Astragalus Radix ＋ Angelicae Sinensis Radix 1：1 group. These results suggested that t-BHP can promote HSCs senescence and reduce cell proliferation ability. Angelicae Sinensis Radix, Astragalus Radix and their combinations can inhibit HSCs senescence, promote HSCs proliferation as well as cell cycle conversion ; moreover, the effects of 1 ： 1 Astragalus Radix ＋ Angelicae Sinensis Radix were strongest. The mechanisms may be related to up-regulating the expression of cell cycle positive regulator, down-regulating the expression of cell cycle negative regulator, thus promoting the cells to ent

关 键 词：造血干细胞 衰老 细胞增殖 黄芪 当归 细胞周期 

分 类 号：R285.5[医药卫生—中药学]