益气养阴方及其拆方影响急性髓细胞白血病细胞Flt3和N-ras表达研究

作者	单位
徐瑞荣	山东中医药大学附属医院血液科
刘奎	山东中医药大学附属医院血液科
王晓玲	山东中医药大学附属医院血液科
胡述博	山东中医药大学附属医院血液科
崔兴	山东中医药大学附属医院血液科
王琰	山东中医药大学附属医院血液科
王敬毅	山东中医药大学附属医院血液科
李丽珍	山东大学齐鲁医院免疫室

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中文摘要:

目的研究益气养阴方及其拆方后的扶正方、祛邪方对Flt3和N-ras在人急性髓细胞白血病(acute myeloid leukemia,AML)表达的影响,探讨益气养阴方治疗白血病的作用机制。方法收集60例AML患者骨髓单个核细胞,将所提取每例患者的细胞混悬液分成4组:对照组不加药物,实验组分别加入益气养阴方、扶正方、祛邪方中药制剂。采用逆转录-聚合酶链反应(RT-PCR),免疫印迹法(Western bloting)观察益气养阴组、扶正组、祛邪组及对照组对Flt3、N-ras基因及FLT3蛋白表达的影响。结果 RT-PCR法检测:对照组、益气养阴组、扶正组、祛邪组Flt3基因表达率分别为(90.78±6.92)%、(38.18±4.50)%、(65.57±5.55)%、(61.35±6.39)%,N-ras基因表达率分别为(93.28±5.54)%、(34.38±6.69)%、(59.42±7.35)%、(65.28±7.64)%,益气养阴组、扶正组、祛邪组与对照组比较,差异均有统计学意义(P<0.05)。Westernbloting检测:对照组、益气养阴组、扶正组、祛邪组FLT3蛋白灰度值分别为0.8127±0.0284、0.4265±0.0353、0.5396±0.0274、0.5473±0.0282,对照组与益气养阴组、扶正组、祛邪组比较差异有统计学意义(P<0.01)。结论中药益气养阴方可抑制AML细胞的克隆性增殖,可降低Flt3和N-ras在AML细胞的表达水平,对AML治疗具有作用。

英文摘要:

Objective To study the action mechanism of Yiqi Yangyin Recipe (YYR) in treating leukemia by observing the effects of YYR and its different assembling,energy supporting part (P1) and evil dispelling part (P2) on expressions of Flt3 and N-ras gene in acute myeloid leukemic (AML) cells. Methods The mononuclear cells collected from bone marrow of 60 AML patients were assigned to four groups:the blank was untreated for control and the three tested groups were treated with YYR,P1 and P2,respectively. The effects on Flt3 and N-ras gene expressions and FLT3 protein expression were observed by RT-PCR and Western bloting. Results RT-PCR test showed the expression of Flt3 in the control,YYR,P1 and P2,group was 90.78%±6.92%,38.18%±4.50%,65.57%±5.55% and 61.35%±6.39%,respectively; and that of N-ras in them 93.28%±5.54%,34.38%±6.69%,59.42%±7.35% and 65.28%±7.64%,respectively,both showed significant difference as compared the data in the tested groups with those in the control group (P<0.05). Western bloting test showed the FLT3 protein gray value in the four groups was 0.8127±0.0284,0.4265±0.0353,0.5396±0.0274 and 0.5473±0.0282,respectively,also showed significant difference between the control and the tested groups (P<0.01). Conclusion YYR can inhibit the colonic proliferation of AML cells,decrease the expressions of FLT3 and N-ras in cells,therefore shows a therapeutic effect on AML.

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