谭骏岚;任丽芝;龙梦铃;李宁;林宏远;梁慧;

• 1:湖南中医药大学
• 2:湖南省肿瘤医院

摘要(Abstract)：

目的 观察肠复方含药血清干预后的肠癌髓源性抑制细胞(MDSCs)对CD8⁺ T细胞功能的影响，探究其可能作用机制。方法 建立小鼠结肠癌皮下移植瘤模型，免疫磁珠法分选小鼠脾脏MDSCs及外周血CD8+ T细胞功能的影响，探究其可能作用机制。方法 建立小鼠结肠癌皮下移植瘤模型，免疫磁珠法分选小鼠脾脏MDSCs及外周血CD8⁺ T细胞并鉴定活性及纯度。将20只SD大鼠随机分为空白血清组、香菇多糖组及肠复方低、中、高剂量组，每组4只，除空白血清组外，其余组予以相应药物灌胃制备含药血清。采用各组大鼠血清培养小鼠脾脏MDSCs,一部分用流式细胞术检测活性氧(ROS)水平，Western blot法检测gp91phox和p47phox蛋白表达水平；另一部分MDSCs与CD8+ T细胞并鉴定活性及纯度。将20只SD大鼠随机分为空白血清组、香菇多糖组及肠复方低、中、高剂量组，每组4只，除空白血清组外，其余组予以相应药物灌胃制备含药血清。采用各组大鼠血清培养小鼠脾脏MDSCs,一部分用流式细胞术检测活性氧(ROS)水平，Western blot法检测gp91phox和p47phox蛋白表达水平；另一部分MDSCs与CD8⁺ T细胞建立共培养体系，采用ELISA法检测共培养体系上清液中白细胞介素-2(IL-2)、干扰素γ(IFN-γ)水平。结果 免疫磁珠分选后的MDSCs纯度为91.35%,CD8+ T细胞建立共培养体系，采用ELISA法检测共培养体系上清液中白细胞介素-2(IL-2)、干扰素γ(IFN-γ)水平。结果 免疫磁珠分选后的MDSCs纯度为91.35%,CD8⁺ T细胞为99.78%,两者活性均大于90%。肠复方中、高剂量组肠癌MDSCs中ROS水平、gp91phox和p47phox蛋白表达水平均明显低于空白血清组(P均<0.05),且肠复方高剂量组均明显低于肠复方中剂量组(P均<0.05);香菇多糖组、肠复方低剂量组肠癌MDSCs中ROS水平、gp91phox和p47phox蛋白表达水平虽较空白血清组有下降趋势，但差异均无统计学意义(P均>0.05)。香菇多糖组及肠复方低、中、高剂量组共培养体系上清液中IL-2、IFN-γ水平均明显高于空白血清组(P均<0.05),肠复方低、中、高剂量组均明显高于香菇多糖组(P均<0.01),且肠复方高剂量组均明显高于肠复方低、中剂量组(P均<0.05)。结论 肠复方可以逆转肠癌MDSCs对CD8+T细胞的抑制作用，其机制可能与抑制gp91phox和p47phox蛋白表达而降低MDSCs中ROS的水平有关。

关键词(KeyWords)： 肠癌;肠复方;髓源性抑制细胞;CD8~+ T细胞;免疫功能

Abstract:

Keywords:

基金项目(Foundation): 湖南省中医药管理局重点项目(201807);; 湖南省自然科学基金面上项目(2017JJ2175);; 长沙市科技局项目(kp1701044)

作者(Authors): 谭骏岚;任丽芝;龙梦铃;李宁;林宏远;梁慧;

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