【摘要】 目的:研究氧化苦参碱(MOX)对Colon26肿瘤细胞免疫抑制的体外影响.方法:获取体外经MOX作用后再培养的Colon26培养上清,以不经MOX作用的Colon26同步培养上清作对照,观察其对小鼠脾细胞MTT法测定的NK杀伤和ConA诱导转化以及流式细胞计数分析的IL-2Rα,CD3ε+ζ+和CD3ε-ζ+表达5项免疫功能指标的影响,定量ELISA法测定这些上清中TGF-β1,VEGF,IL-4,IL-6和IL-105种免疫抑制分子的含量,分析MOX下调Colon26分泌免疫抑制分子与上清免疫抑制作用的关系.结果:不经MOX作用同步培养的Colon26上清,5种免疫抑制分子均可被测到,以TGF-β1含量最高;该上清对小鼠脾细胞5项免疫功能指标,均有显著抑制作用.MOX作用后的Colon26,其第一次再培养上清,TGF-β1和IL-10含量及对除NK杀伤以外的其余4项免疫功能指标的抑制,均明显降低(P值分别为0.0045,0.032,0.001,0.0095,0.0005,0.000);与第一次再培养上清相比,其第二次再培养上清,TGF-β1含量继续显著降低(P=0.002),VEGF含量及对除NK杀伤以外的其余4项免疫功能指标的抑制,均明显提高(P值分别为0.029,0.0075,0.0495,0.0005,0.0005),其余无显著变化.结论:MOX可明显下调Colon26肿瘤细胞TGF-β1和IL-10的分泌,通过下调所测5种以外的其他免疫抑制分子的分泌而影响Colon26肿瘤细胞的免疫抑制效应.下调肿瘤细胞免疫抑制作用,可能是MOX的抗瘤效应机制之一.更多还原

【Abstract】 AIM:To study in vitro the down-regulating effects of Matrine N-Oxide on immunosuppression of Colon26 tumor cells. METHODS:Re-cultured supernatants of Colon26 after treated by Matrine N-Oxide and the corresponding supernatants of Colon26 treated without Matrine N-Oxide as a control group were collected. The effects of different supernatants on 5 immune functions of murine splenocytes were determined, including NK killing and transformation induced by ConA detected by MTT, and expression levels of IL-2Rα, CD3ε⁺ζ⁺ and CD3ε^-ζ⁺ detected by FCM. The concentrations of 5 immunosuppressive molecules （including TGF-β₁, VEGF, IL-4, IL-6 and IL-10） in different supernatants were measured by quantitative ELISA. The relationships between the down-regulating effect of Matrine N-Oxide on the secretions of immunosuppressive molecules and the immunosuppression of Colon26 supernatant were analyzed. RESULTS:For supernatants of Colon26 treated without Matrine N-Oxide, all of the 5 immunosuppressive molecules were found, the concentration of TGF-β₁ was highest, and the significant inhibitions of the 5 immune functions of murine splenocytes were showed. For Colon26 after treated by Matrine N-Oxide, the concentrations of TGF-β₁ and IL-10 in the first re-cultured supernatant and its inhibitions of the 4 immune functions except of NK killing decreased greatly （P= 0.0045, 0.032, 0.001, 0.0095, 0.0005, 0.000, respectively）. Compared with the first re-cultured supernatant, the concentrations of TGF-β₁ in the second re-cultured supernatant continued decreasing （P=0.002）, the concentrations of VEGF and the inhibitions of the 4 immune functions except of NK killing increased highly （P=0.029, 0.0075, 0.0495, 0.0005, 0.0005, respectively）, and others had no change. CONCLUSION:Matrine N-Oxide can down-regulate remarkably the secretions of TGF-β₁ and IL-10 by Colon26 and its reversion of tumor immunosuppression may be through down-regulating other immunosuppressive molecules except that detected in this study. Down-regulating the immunosuppressions of tumor cells should be one of anti-tumor mechanisms of Matrine N-Oxide.更多还原