当归多糖、黄芪多糖及其配伍对化疗性骨髓抑制小鼠骨髓干细胞凋亡和脱核因子的影响  

Effects of Angelica Polysaccharides,Astragalus Polysaccharides and Their Compatibility on Apoptosis and Denucleation Factors of Bone Marrow Stem Cells in Mice with Chemotherapy-induced Bone Marrow Suppression

作　　者：崔运浩[1] 初杰[1] 范颖[1] 李新[1] 蒋宁[1] 林庶茹[1] 张红梅[1] 李影迪 李秀[1] CUI Yunhao;CHU Jie;FAN Ying;LI Xin;JIANG Ning;LIN Shuru;ZHANG Hongmei;LI Yingdi;LI Xiu(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学

出　　处：《辽宁中医药大学学报》2019年第8期38-43,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省自然科学基金项目(20170540611)

摘　　要：目的:通过实验研究当归多糖、黄芪多糖及其配伍对化疗性骨髓抑制小鼠骨髓干细胞细胞凋亡因子BCL-2、Caspase9、Bax,脱核因子C-myc、HDAC2的影响。方法:C57BL/6小鼠20只,雌雄各半,28~32g。给模型组小鼠每日腹腔注射一次环磷酰胺380mg·kg^-1,正常小鼠腹腔注射等量生理盐水,连续3d。造模后,以脱颈椎法处死各组小鼠,取出股骨,剔除肌肉和结缔组织,剪开股骨两端,用6号针头以生理盐水反复冲洗骨髓腔,并将冲出的骨髓打碎,再用4号针头过滤制成单个细胞悬液,在离心机上2000r/min,离心10min,去上清液,加入4mL冷70%乙醇固定,上振荡器摇匀制成骨髓干细胞样本。细胞实验分为6组,包括正常组、模型组、“EPO”组、“EPO+(G-CSF)”组、当归多糖组、黄芪多糖组、两药配伍组。直接给药当归多糖0.2mg/mL,黄芪多糖0.2mg/mL,EPO50U/mL,G-CSF50U/mL于骨髓干细胞体外培养体系中,培养3d,室内18~20℃,20%相对湿度,5%CO2,37℃孵育箱。采用RT-qPCR法检测骨髓干细胞中BCL-2、Caspase9、Bax、C-myc、HDAC2的mRNA表达。结果:(1)造模后模型组小鼠骨髓干细胞BCL-2mRNA表达下降,Caspase9、BaxmRNA表达升高,与正常组比较差异均有统计学意义(P<0.05)。①各给药组与模型组BCL-2mRNA均有明显差异,当归多糖有提升BCL-2mRNA作用,黄芪多糖作用不明显,两药间有交互作用,合用不如单一用药。②各给药组与模型组Caspase9mRNA均有明显差异,当归多糖与黄芪多糖均有降低Caspase9mRNA作用,两药交互作用明显,合用不如单用黄芪多糖。③各给药组与模型组BaxmRNA均有明显差异,黄芪多糖有降低BaxmRNA作用,当归糖作用不明显,两药交互作用无统计学意义。(2)造模后模型组小鼠骨髓干细胞C-myc、HDAC2mRNA表达升高,与正常组比较差异均有统计学意义(P<0.05)。①各给药组与模型组C-mycmRNA均有明显差异,当归多糖与黄芪多糖均有降低C-mycmRNA作用,两药交互作用明显,�Objective:To study the effects of Angelica polysaccharides,Astragalus polysaccharides and their compatibility on apoptotic factors BCL-2,Caspase9,Bax,denucleation factors C-myc and HDAC2 of bone marrow stem cells in mice with chemotherapy-induced bone marrow depression.Methods:Twenty C57BL/6 mice,half male and half female,28-32 g.The mice in the model group were given intraperitoneal injection of cyclophosphamide 380 mg kg^-1 once a day,and the normal mice were given intraperitoneal injection of normal saline for 3 consecutive days.After modeling,the mice in each group were killed by cervical decortication.The femur was taken out,the muscles and connective tissue were removed,the ends of femur were cut off,the marrow cavity was washed repeatedly with No.6 needle with normal saline,and the washed marrow was broken.Then the washed marrow was filtered with No.4 needle to form a single cell suspension.The cell suspension was centrifuged for 2000 r/min,centrifuged for 10 min,the supernatant was removed,fixed with 4 mL cold 70%ethanol,and shaken by an oscillator to make the bone marr cell samples.Cell experiments were divided into six groups,including normal group,model group,EPO group,EPO+(G-CSF)group,Angelica polysaccharide group,Astragalus polysaccharide group,two drug compatibility group.Directly administered Angelica polysaccharide 0.2 mg/mL,Astragalus polysaccharide 0.2 mg/mL,EPO 50 U/mL,and(G-CSF)50 U/mL in vitro culture system of bone marrow stem cells for 3 days,incubation chamber at 18-20 g,20%relative humidity,5%CO2,37℃.The expression of BCL-2,Caspase9,Bax,C-myc and HDAC2 in bone marrow stem cells was detected by RT-q PCR.Results:(1)The expression of BCL-2 and Caspase9 and Bax in bone marrow stem cells of mice in the model group decreased and increased,with significant difference compared with the normal group(P<0.05).①There were significant differences in BCL-2 mRNA between each drug group and model group.Angelica polysaccharides could enhance BCL-2 mRNA,but astragalus polysaccharides had no obvious eff

关 键 词：当归多糖 黄芪多糖 BCL-2 CASPASE9 Bax C-myc HDAC2 

分 类 号：R285.5[医药卫生—中药学]