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目的:研究红芪多糖对S180瘤细胞化疗协同增效作用的差异蛋白质。方法:以环磷酰胺和红芪多糖联合环磷酰胺治疗S180荷瘤小鼠14天后测定小鼠免疫器官指数及抑瘤率,利用双向电泳分离提取的瘤细胞总蛋白,考染显色后应用PDQuest8.0软件分析电泳图谱,找出差异表达的蛋白质。结果:与环磷酰胺(20 mg/kg)组相比,红芪多糖(200 mg/kg)联合环磷酰胺(20 mg/kg)可明显抑制小鼠S180瘤生长(P<0.05),显著减轻环磷酰胺所致免疫器官指数的下降(P<0.01);建立了背景清晰、重复性好、分辨率高的S180瘤细胞双向电泳图谱,环磷酰胺组与红芪多糖联合环磷酰胺组蛋白质点数分别为:776±22和721±16;平均匹配率分别为:91.13%和89.37%。比较分析环磷酰胺组与红芪多糖联合环磷酰胺组双向电泳图谱,筛选出差异明显的蛋白质点28个,其中8个表达上调,19个表达下调,1个仅在环磷酰胺组表达。结论:红芪多糖联合环磷酰胺可减轻其对S180荷瘤小鼠的免疫抑制作用,增强环磷酰胺对S180瘤细胞的化疗效果,其作用可能与筛选出的28个差异蛋白质点有关,对这些差异蛋白质点的鉴定将为揭示红芪多糖的化疗协同增效作用提供实验依据。
OBJECTIVE: To study the differential proteins of synergistic effect of Radix Hedysari Polysaccharides on S180 tumor cells. Methods: S180 tumor-bearing mice were treated with cyclophosphamide and Hedysari polysaccharide combined with cyclophosphamide for 14 days. Immuno-organ index and tumor inhibition rate of mice were measured 14 days later. The total protein of tumor cells was separated by two-dimensional electrophoresis. PDQuest8.0 software analysis of electrophoresis patterns to identify differentially expressed proteins. Results: Compared with cyclophosphamide (20 mg / kg), the combination of 300 mg / kg HCH and cyclophosphamide (20 mg / kg) significantly inhibited the growth of S180 tumor in mice (P <0.05) (P <0.01). A two-dimensional gel electrophoresis pattern of S180 tumor cells with clear background, good repeatability and high resolution was established. Cyclophosphamide group and Radix Hedysari combined with cyclophosphamide group The protein spots were 776 ± 22 and 721 ± 16, respectively. The average match rates were 91.13% and 89.37% respectively. The two-dimensional gel electrophoresis of cyclophosphamide and Radix Hedysari combined with cyclophosphamide was compared to screen out 28 proteins with obvious difference, of which 8 were up-regulated, 19 down-regulated, and 1 was only expressed in cyclophosphamide. Conclusion: Radix Hedysari combined with cyclophosphamide can reduce its immunosuppressive effect on S180 tumor-bearing mice and enhance the chemotherapeutic effect of cyclophosphamide on S180 tumor cells, which may be related to the screening of 28 differential protein spots. The identification of these differential proteins will provide experimental evidence for revealing the synergistic effect of Radix Hedysari.