当归/黄芪多糖对腹腔注射环磷酰胺小鼠骨髓造血影响随机平行对照研究

Angelica Sinensis/Astragalus Polysaccharides on Hematopoietic Function of Bone Barrow of Mice Induced by Intraperitoneal Injection of Cyclophosphamide impact a randomized parallel controlled study

[目的]观察当归/黄芪多糖对腹腔注射环磷酰胺小鼠骨髓造血影响。[方法]使用随机平行对照方法,将60只清洁级C57BL/6小鼠编号按随机数字表法随机分为6组,空白组、模型组、促红细胞生成素组(EPO组)、当归多糖组(当归组)、黄芪多糖组(黄芪组)、当归加黄芪多糖组(当归+黄芪组),10只/组。实验第1d,各组小鼠称体重,模型组、EPO组、当归组、黄芪组、当归+黄芪组,腹腔注射环磷酰胺380mg/kg·d^(-1),连续3d,空白组注射等量生理盐水。实验第4d(造模第4d),颈椎脱臼处死小鼠,无菌取股骨和胫骨,置入培养基中,调整细胞浓度为1×10~5/m L,然后将混合液加入96孔板(200u L)/孔和24孔板(1L/孔)中,每组设3个复孔,周围孔分别加入相应量的无菌超纯水,将细胞板置于5%CO_2,37℃孵育箱中培养。实验第4d(造模第4d),各组分别干预。观测外周血象、细胞增殖率(MTT法),EPO、IL-3(免疫蛋白法),细胞内转录因子JAK2、STAT5(RT-PCR法)。[结果]细胞培养24h、48h和72h,细胞增殖率EPO组、当归组、黄芪组、当归+黄芪组均高于空白组(P<0.05,P<0.01),组间无显著差异(P>0.05);模型组低于空白组(P<0.05,P<0.01)。细胞培养7d,IL-3、EPO表达模型组低于空白组(P<0.01),各干预组均高于模型组(P<0.01),当归+黄芪组高于EPO组(P<0.01),EPO组高于当归组(P<0.01),当归组高于黄芪组(P<0.01)。细胞培养7d,单核细胞STAT5、JAK2m RNA模型组低于空白组(P<0.01),各干预组高于模型组(P<0.05或P<0.01),当归+黄芪组高于EPO组(P<0.01),EPO组与当归组无显著差异(P<0.05),当归组高于黄芪组(P<0.01)。[结论]当归/黄芪多糖、促红细胞生成素干预腹腔注射环磷酰胺小鼠,均可升高骨髓细胞增殖率、IL-3、EPO表达及单核细胞STAT5、JAK2m RNA,当归多糖与黄芪多糖联合应用更明显。

[objective] To observe the effect of angelica sinensis/astragalus polysaecharides on hematopoietic fimction of bone harrow of mice induced by intraperitoneal injection of cyclophosphamide. [Methods] The randomized parallel control method was used, 60 clean C57BL/6 mice were divided into six groups in a randomized manner by random number table of normal group, model group, Erythropoietin （EPO） group, angelica sinensis polysaccharides （angelica sinensis） group, astragalus polysaccharides （astragalus） group, angelica sinensis and astragalus polysaccharides （angelica sinensis＋astragalus） group, with 10 mice in each group. By referring the blood-deficient model copy method, in the first clay of the experiment, mice of various groups were weighted, intraperitoneal injection of 380mg/Kg ·d-1 of cyclophosphamide was performed in model group, EPO group, angelica sinensis group, astragalus group, angelica sinensis and astragalus group for three successive days, while equivalent normal saline was given to normal control group. In the fourth day ofexperiment （molding）, the mice were killed by cervical dislocation, femur and tibia were taken in a sterile way, then put in a culture medium, the cell concentration was adjusted to 1 ×10^5/mL, the mixed liquor was added to 96-pore plate （200 uL/pore） and 24-pore plate （1L/pore）, 3 double-pores were equipped for each group, a certain amount of sterile ultrapure water was added respectively to each surrounding pore, then the cell plate was placed into a 5%CO2, 37℃ incubator for culture. In the fourth day of experiment （molding）, interference measures were taken for each group. The peripheral blood, cell proliferation rate （MTY）, EPO, IL-3 （ELLISA）, ceil transcription factor JAK2, STATS （RT-PCR） were observed. [Results] after cells were cultured for 24h, 48h and 72h respectively, the cell proliferation rate of EPO group, angelica sinensis group, astragalus group, angelica sinensis and astragalus group was respectively higher than that of normal group （P〈0.05, P〈0.01）, no significant difference among groups （P〉0.05）; while that of model group was lower than that in normal group （P〈0.05, P〈0.01）. After the cells were cultured for 7d, IL-3 and EPO expression for model group was lower than that for normal group （P〈0.01）, that for each interference group was higher than that for model group （P〈0.01）, that for angelica sinensis and astragalus group was higher than that for EPO group （P〈0.01）, that for EPO group was higher than that for angelica sinensis group （P〈0.01）, and that for angelica sinensis group was higher than that for astragalus group （P〈0.01）. After cells were cultured for 7d, monocyte STAT5 and JAK2Mma for model group were lower than those for normal group （P〈0.01）, those for each interference group were higher than those for model group （P〈0.05, P〈0.01）, those for angelica sinensis and astragalus group were higher than those for EPO group （P〈0.01）, with no significant differences between EPO group and astragalus group （P〈0.05）, those for angelica sinensis were higher than those for astragalus group （P 〈0.01）. [Conclusion] the use of angelica sinensis/astragalus polysaccharides and erythropoietin to intervene in mice with intraperitoneal injection of cyclophosphamide can increase bone barrow proliferation rate, il-3, EPO expression and monocyte STAT5 and JAK2mRNA, and combined application of angelica sinensis and astragalus polysaccharides will have a more apparent curative effect.