三七皂苷对人骨髓造血细胞凋亡相关蛋白表达影响.docVIP

三七皂苷对人骨髓造血细胞凋亡相关蛋白表达影响 　 ; 作者：陈小红，高瑞兰，郑智茵，钱煦岱，徐卫红 毕业论文 【摘要】; 本文研究观察三七皂苷（PNS）对造血细胞促进凋亡相关蛋白（Daxx、Fas）和抑制凋亡的核转录因子（NFkB、c-Rel）表达的影响，探讨PNS促进造血细胞增殖，支持生长存活的作用机制。采用半固体集落培养法观察PNS对人骨髓红系、粒系祖造血细胞（CFU-GM、CFU-E）的增殖作用，台盼蓝拒染法观察细胞的存活率，涂片染色法观察细胞形态学的变化，用流式细胞术分析细胞凋亡状况。同时，用PNS 处理粒系HL-60、红系K562、巨核系CHRF-288和Meg-01 4种细胞株后，提取胞浆蛋白和核蛋白，Western 免疫印迹观察Daxx、Fas、NFkB和c-Rel蛋白的表达水平。结果发现： ①PNS能够刺激人骨髓红系、粒系祖细胞和HL-60等4种细胞株增殖。②HL-60等4种株细胞经PNS和饥饿处理后，活性良好，镜下未观察到凋亡的形态学特征；AnnexinⅤ分析也未见凋亡的阳性细胞。③Western blot结果显示，经PNS处理的4株细胞Daxx蛋白表达水平与对照相比，下降幅度分别为33.3%-61.5%；HL-60细胞本身表达Fas蛋白低下，PNS处理后也无明显下降，而K562、CHRF-288和Meg-01细胞Fas蛋白表达与对照相比，下降幅度分别为33.3%-71.4%。④ NFkB、c-Rel蛋白除HL-60细胞对PNS诱导无明显变化外，其余细胞均出现不同程度地增加，分别提高了2.0-2.7倍和1.5-2.3倍。结论： PNS在某种程度上能够抑制Daxx、Fas蛋白表达，而相应减少造血细胞的凋亡，同时也能通过上调NFkB、c-Rel转录因子，促进细胞增殖，并阻止半胱天冬酶（caspase）连锁链的活化而抑制造血细胞凋亡，这为PNS应用于临床治疗凋亡过度的疾病如再生障碍性贫血提供了可能性。 毕业论文 【关键词】; 造血细胞 毕业论文 　　Expression of Apoptosis-related Proteins in the Human Bone Marrow Hematopoietic Cells Treated by Panax Notoginosides 毕业论文 　　Abstract The study was aimed; to investigate the action of Panax Notoginosides; (PNS，extracted from notoginseng herb) on the expression of the apoptosis-related proteins (Daxx，Fas) and transcription factors (NFkB、c-Rel) in the hematopoietic cells and to; explore the mechanisms of supporting cells to survive.The colony formation of CFU-GM and CFU-E in human bone marrow; was assayed in the presence of various; concentrations of PNS.The viability of cells was; assayed; by trypan blue and the changes of cell morphology were observed with microscope.The Annexin-V positive cells were detected by FCM.Three lineages; of human myeloid HL-60，erythroid K562，megakaryoid CHRF-288 and Meg-01 cells were incubated in addition; of PNS（10 mg/L）for 14 days.The nuclear or cytoplasm protein of cells was extracted and analyzed by Western blot with monoclonal antibodies against Daxx，Fas or NFkB，c-Rel.The results showed: (1)the proliferation on hematopoietic progenitor cells （CFU-GM and CFU-E） and four cell lines was promoted