当归注射液对免疫诱导再生障碍性贫血小鼠CD34＋细胞及其Fas抗原表达的影响

Effects of angelica sinenisis injection on CD_(34)～+ cells and CD_(34)～+-Fas antigen in immune-induced aplastic anemia mice

背景:前期实验已证实当归注射液能改善再生障碍性贫血小鼠骨髓微环境,促进造血细胞增生。能增加细胞周期蛋白D2表达,促进细胞从G1→S期的转换。目的:拟证实当归注射液对免疫诱导再生障碍性贫血小鼠CD34+细胞及其Fas抗原表达的影响。设计、时间和单位:完全随机区组设计的细胞分子学实验,于2005-11/2006-04在武汉大学人民医院儿科研究所进行。材料:选用8～12周龄雌性(H-2a、MLSb)Balb/c小鼠作为受体,制作再生障碍性贫血模型。选用DBA/2小鼠(H-2a、MLSa)8～10周龄作为供体。方法:将雌性Balb/c小鼠随机分为3组:对照组、模型组和治疗组,每组8只。模型组和治疗组动物建立再生障碍性贫血模型。对照组不经照射。于造模当天开始,模型组和对照组腹腔注射无菌生理盐水10mL/(kg·d),治疗组腹腔注射当归注射液10mL/(kg·d),1次/d,连续12d。主要观察指标:于第12天每只小鼠采集尾静脉血测外周血白细胞计数,取出股骨,计数骨髓单个核细胞。酶联免疫法检测肿瘤坏死因子α表达水平,经流式细胞仪检测骨髓单个核细胞CD34+细胞及CD34+细胞Fas抗原表达。结果:模型组和治疗组小鼠外周血白细胞及骨髓单个核细胞计数均明显低于对照组,治疗组明显高于模型组(P<0.01)。模型组CD34+抗原表达水平明显低于对照组,治疗组CD34+抗原表达水平明显高于模型组(P<0.01),已接近对照组水平。对照组CD34+细胞Fas抗原表达最低,模型组Fas抗原表达明显升高,与对照组比较差异有显著性意义(P<0.01),表明细胞增殖受影响。治疗组Fas抗原的表达低于模型组(P<0.01)。模型组小鼠肿瘤坏死因子α表达明显高于对照组,治疗组明显低于模型组(P<0.01)。结论:当归注射液可能通过降低负调控因子肿瘤坏死因子α水平,减少细胞凋亡,促进再生障碍性贫血小鼠造血细胞的增殖。

BACKGROUND： Previous studies have confirmed that angelica sinenisis injection can improve bone marrow microenvironment, promote hematopoietic cellular proliferation, increase the expression levels of cyclin D2, and promote the transform of hematopoietic cell from G1 to S phase in aplastic anemia mice. OBJECTIVE： To investigate the influence of angelica sinenisis injection on CD34＋ cells and CD34＋-Fas antigen in immune-induced aplastic anemia mice. DESIGN, TIME AND SETTING： A randomized control animal experiment was performed at the Institute of Pediatrics, Renmin Hospital, Wuhan University from November 2005 to April 2006. MATERIALS： 8-12 week old female （H-2a , MLSb） Balb/c mice acted as receptors were used to make aplastic anemia models. 8-10 week old DBA/2 （H-2a, MLSa） mice served as donors. METHODS： Female Balb/c mice were randomly divided into control group, model group and therapy group, with 8 rats for each group. Mice of the model and therapy groups were made into aplastic anemia models. Mice of the control group were not irradiated. After establishing aplastic anemia models, each mouse of the model and control groups was intraperitoneally injected with sterile saline 10 mL/kg per day for 12 days. Mice of the therapy group were intraperitoneally injected with angelica sinenisis injection 10 mL/kg per day, once a day, for 12 days. MAIN OUTCOME MEASURES： At 12 days, peripheral blood leucocytes were counted via the tail vein from each mouse, and then the femur was taken out. The number of bone marrow mononucleate cells was measured. Tumor necrosis factor （TNF）- α expression was measured by euzymelinked immunosorbent assay. CD34＋ cells and CD34＋-Fas antigen expression were measured by the flow cytometry. RESULTS： Compared with the control group, the number of peripheral blood leucocytes and mononuclear cells was obviously reduced in the model and therapy groups, but significantly higher in the therapy group than in the model group （P〈 0.01）. The expression levels of CD34＋ antigen in the model group were significantly lower than in the control group, and significantly higher in the therapy group than in the model group （P〈 0.01）, nearly to the levers of the control group. CD34＋-Fas antigen expression was the lowest in the control group, significantly increased in the model group, and it has significant difference compared with the control group （P 〈 0.01）. These indicated that the cell proliferation was influenced. CD34＋-Fas antigen expression in the therapy group was lower than in the model group （P 〈 0.01）. The expression of TNF-α in the model group were significantly higher than in the control group, but lower in the therapy group than in the model group （P 〈 0.01）. CONCLUSION： Perhaps angelica sinenisis injection can promote the proliferation of hematopoietic cells in aplastic anemia mice by reducing the expression of TNF- α and apoptosis.