淫羊藿提取物对急性髓系白血病SCID小鼠模型细胞生物学特性的影响

摘要

目的 观察淫羊藿提取物对急性髓系白血病SCID模型小鼠细胞生物学特性的影响.方法 SCID小鼠共60只,随机选取其中的50只,采用腹腔注射对数生长期HL-60细胞株于经环磷酰胺预处理的SCID小鼠以建立急性髓系白血病SCID小鼠模型,4周后随机均分为模型组、阿糖胞苷组、观察1组、2组和3组,另10只正常小鼠设为空白对照组.观察1组、2组和3组分别用淫羊藿提取物10、5、2.5 mL/kg灌胃治疗5周,模型组和空白对照组灌胃等体积0.9%氯化钠注射液5周,阿糖胞苷组尾静脉注射阿糖胞苷.在治疗第5周时尾静脉采血,采用人工法计数外周血血象及分类;采用Western blotting法检测同源性磷酸酶-张力蛋白(PTEN)和多聚腺苷二磷酸核糖多聚酶(PARP)蛋白表达;流式细胞学检测HL-60细胞膜CD13和CD71表达率及细胞各周期细胞百分率.结果 观察2组、3组外周血白细胞、淋巴细胞和HL-60细胞总数明显降低,中性粒细胞升高,PARP蛋白表达量、CD13和CD71阳性表达率明显下降,PTEN蛋白表达量增加,与模型组比较差异有统计学意义(P<0.05).两组HL-60细胞均被阻滞于G1期,且G1和S期细胞百分率呈浓度效应关系.结论 淫羊藿提取物有与阳性药阿糖胞苷相似的药理作用,在体内对急性髓系白血病HL-60细胞株有一定的抑制作用,其机制可能通过下调CD13和CD71,裂解PARP以诱导急性髓系白血病细胞凋亡,增强PTEN蛋白表达并促使细胞阻滞于G1期,从而发挥抑制白血病细胞的增殖作用.%Objective: To observe the effects of epimedium herb extracts on the cell biological characteristics of SCID mouse models with acute myeloid leukemia. Methods: 50 of SCID mice were randomly selected from 60 ones.SCID mouse models with acute myeloid leukemia were established by intraperitoneal injection of logarithmic phase-HL 60 cells in SCID mice of cyclophosphamide pretreatment. After 4 weeks,they were randomly divided into the model group,cytarabine group,the observation groups: Group 1,Group 2 and Group 3,another 10 normal mice as the blank control group.The observation groups:Group 1,Group 2 and Group 3 were treated for 5 weeks with 10 mL/kg,5 mL/kg and 2.5 mL/kg of physiological saline solution for 5 weeks,respectively.The model group and the blank control group were intragastric with equal volume of normal saline for 5 weeks. Cytarabine group received caudal intravenous injection of cytarabine.In the 5th week of treatment,caudal vein blood was collected. Artificial method was used to count peripheral blood hemogram and classification. Western blotting method was used to detect homologous protein phosphatase-tension(phosphatase and tensin homolog,PTEN)and the protein expression of[poly(ADP-ribose)polymerase;PARP]. FCM was used to detect the expression rate of CD13 and CD71 in hl-60 cell membrane and the percentage of cell and cycling cell. Results: In Group 2 and Group 3,the total number of white blood cells,lymphocytes and HL-60 cells in peripheral blood decreased;neutrophils in-creased;PARP protein expression and positive expression rate of CD13 and CD71 decreased,and the protein ex-pression of PTEN increased,and compared with the model group,the difference was statistically significant(P<0.05). HL-60 cells of the two groups were blocked in the G1phase,and the percentage of G1and S cells showed a concentration effect relationship. Conclusion: The pharmacological action of Epimedium extract is similar to that of cytarabine in positive drugs.Epimedium extract in the body has certain inhibitory effecton HL 60 cell lines of acute myeloid leukemia,and its mechanism may be to induce apoptosis in acute myeloid leukemia cells by downregulation of CD13 and CD71,and lysis PARP,enhance PTEN protein expression and promote cell arrest in G1 phase,thereby inhibiting the proliferation of leukemic cells.