补肾益髓生血法AA大鼠含药血清对BMSCs与BMNCs共培养BMNCs增殖分化及SDF-1/CXCR4/PI3K/AKT信号通路的影响  

Effects of Bushen Yisui Shengxue method for medicated serum of rats with aplastic anemia on proliferation of BMNCs co-culturing with BMSCs and PI3K/AKT signaling pathway mediated by SDF-1/CXCR4

作　　者：程明秀[1] 赵宗江[1] 田晨[1] 吴阿敏[1] 徐昌君[1] 黄雅薇 苗永辉[1] 杨冠男[1] 吴志奎[2] 

机构地区：[1]北京中医药大学,北京100029 [2]中国中医科学院广安门医院,北京100053

出　　处：《中华中医药杂志》2015年第8期2877-2881,I0001,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点基础研究发展计划(973计划)(No.2010CB530406)~~

摘　　要：目的:探讨补肾益髓生血法再生障碍性贫血(AA)大鼠含药血清通过SDF-1/CXCR4介导的PI3K/AKT信号通路对骨髓间充质干细胞(BMSCs)与骨髓单个核细胞(BMNCs)共培养BMNCs增殖能力的影响。方法:60Co-γ联合CTX建立再障大鼠模型,各组按文献方法分离血清冻存备用。在分别建立粒系和红系BMSCs与BMNCs共培养体系基础上,用补肾益髓生血法再障大鼠含药血清进行干预。实验分为空白对照组、正常对照组、模型组、康力龙组、滋肾生血组、益髓生血组和温肾生血组。于第4、8、10天分别计数红系集落形成单位(CFU-E)、粒-单核细胞系集落形成单位(CFU-GM)、红系爆式集落形成单位(BFU-E)。共培养10d后,提取细胞总RNA,RT-PCR检测SDF-1、CXCR4、PI3K、AKT与m TOR m RNA的表达。结果:补肾益髓生血法含药血清促进了共培养BMNCs的增殖分化,其集落形成数目显示各治疗组CFU-E、BFU-E、CFU-GM集落形成数量较模型组增加(P<0.05,P<0.01);RT-PCR结果显示康力龙组、滋肾生血组、益髓生血组、温肾生血组SDF-1、CXCR4、PI3K、AKT、m TOR表达较模型组显著升高(P<0.05,P<0.01),集落形成与基因表达均显示,滋肾生血组优于温肾生血组和益髓生血组(P<0.05,P<0.01)。结论:补肾益髓生血法AA大鼠含药血清可以促进BMSCs与BMNCs共培养体系中BMNCs的增殖分化,这可能与SDF-1/CXCR4介导的PI3K/AKT信号通路有关,其中滋肾生血法优于益髓生血法和温肾生血法。Objective： To explore the effects of Bushen Yisui Shengxue （BYS） method for medicated serum of rats with aplastic anemia （AA） on the proliferation of bone marrow mononuclear cells （BMNCs） co-cultured with bone marrow mesenchymal stem cells （BMSCs） and PI3K/AKT signaling pathway mediated by SDF-1/CXCR4. Methods： The rats models were established by 66Co-γ rays and cyclophosphamide. Then we took the drug-containing serum and put them cryopreserved according to the literature. The rat BMSCs can be isolated and cultured by whole bone marrow adherent method. Respectively established myeloid and erythroid culture system to co-culture BMSCs and BMNCs, which were intervened by BYS method for medicated serum of AA rats. And divided into normal control group, the blank control group, model group, stanozolol group, Yi Sui Sheng Xue （YSSX） group, Zi Shen Sheng Xue （ZSSX） group and Wen Shen Sheng Xue （WSSX） group. Counted CFU-E, CFU-GM, BFU-E in 4, 8, 10 days. After 10 days＇ treatment, collected all of the co-culturing cells, TRIZOL method to extract total RNA, RT-PCR detection of SDF-1 and CXCR4, PI3K, and AKT, mTORmRNA expression. Results： Drug-containing serum of AA rats which were treated by the BYS method promoted proliferation of BMNCs co-cultured with BMSCs, the amounts of forming colonies show that, compared with model group, the amounts of CFU-E, BFU-E, CFU-GM in every treatment group were significantly increased （P〈0.01, P〈0,05）. The amounts of CFU-GM, CFU-E BFU-E in ZSSX group were higher than YSSX group and WSSX group （P〈0.01, P〈0.05）; RT-PCR results showed that, compared with model group, the expression of SDF-1, CXCR4, PI3K, AKT, mTORmRNA in stanozolol group, ZSSX group, YSSX group and WSSX group was decreased （P〈0.01, P〈0.05）. The expression of SDF- 1, CXCR4, PI3K, AKT, mTORmRNA in ZSSX group were far higher than WSSX group and YSSX group （P〈0.01, P〈0.05）. Conclusion： BYS method can enhance multiplication capacity of BMNCs during the co-cultu

关 键 词：再生障碍性贫血 补肾益髓生血法 BMSCS BMNCs 共培养 SDF-1/CXCR4 PI3K/AKT/mTOR 

分 类 号：R285.5[医药卫生—中药学]