【目的】 为探讨东方蜜蜂微孢子虫Nosema ceranae感染蜜蜂后的形态学影响、各组织的感染情况及对中肠组织蛋白质的影响，初步探索蜜蜂自身免疫应答调控。【方法】 分别提取中华蜜蜂/意大利蜜蜂实验组与对照组的蜜蜂中肠组织总蛋白，通过SDS-PAGE电泳技术筛选差异蛋白条带，LC-MS/MS质谱分析技术鉴定差异蛋白质，利用半定量和荧光定量RT-PCR技术验证筛选出的差异蛋白质。【结果】 蜜蜂受东方蜜蜂微孢子虫感染后，中华蜜蜂Apis cerana cerana Fabricius和意大利蜜蜂Apis mellifera Ligustica的体重均未出现显著性差异，而二者受感染的中肠组织形态变化均较为明显。其次，对意大利蜜蜂而言，被筛选出的两条较明显的中肠差异表达蛋白质条带经质谱分析共鉴定到35个候选蛋白质。筛选出的两条中华蜜蜂中肠差异蛋白条带经质谱分析技术共鉴定到11个候选蛋白质。经验证，在感染N. ceranae后的第4天，筛选出的意大利蜜蜂精氨酸激酶（AK）和甘油三磷酸脱氢酶（GPDH）及中华蜜蜂硫氧还原蛋白还原酶（TrxR）及精氨酸激酶（AK）的蛋白表达量分别是对照组的2.02倍、2.21倍、1.72倍和1.88倍，均呈现上调表达，与蛋白质差异表达趋势一致。【结论】 在东方蜜蜂微孢子虫感染意大利蜜蜂或中华蜜蜂后，筛选获得差异表达的蛋白质条带并进行鉴定。其中与蜜蜂中肠组织基本代谢、免疫应答、氧化应激与能量代谢相关蛋白质（如意大利蜜蜂的AK和GPDH、中华蜜蜂的AK和TrxR）的表达量被显著上调，暗示这些差异蛋白可能参与了蜜蜂中肠组织的免疫防御。本研究为深入探讨蜜蜂中肠对蜜蜂微孢子虫的免疫与代谢应答方式提供了丰富的数据基础。

[Objectives]  To investigate the morphological effects of infection of honeybees by Nosema ceranae, including the main tissues infected, the effects on protein expression in the midgut and the regulation of the honeybee immune response. [Methods]  Total honeybee midgut proteins were extracted and screened using SDS-PAGE and differentially expressed proteins identified using LC-MS/MS. Differential expression of proteins was verified with semi-quantitative, and quantitative, RT-PCR. [Results]  There was no significant difference in the body weight of infected and uninfected (control) Apis cerana cerana Fabricius or Apis mellifera Ligustica. However, there were significant differences in the intestinal morphology of infected and uninfected bees. Furthermore, two differentially expressed midgut protein bands were identified in A. mellifera and A. cerana cerana. Thirty-five candidate proteins were obtained by MS in A. mellifera and 11 in A. cerana cerana. Four days after infection, expression levels of arginine kinase (AK) and glycerol triphosphate dehydrogenase (GPDH) in A. mellifera, were up-regulated 2.02 times, 2.21 times, respectively, relative to the control group, whereas thioredoxin reductase (TrxR) and arginine in A. cerana cerana were up-regulated 1.72 times, and 1.88 times, respectively. All data were consistent with the results of SDS-PAGE analysis. [Conclusion]  The expression of proteins related to basic metabolism, immune response, oxidative stress and energy metabolism in the honeybee midgut (such as AK and GPDH in A. mellifera and AK and TrxR in A. cerana cerana) were significantly up-regulated after infection by N. ceranae, suggesting that these proteins may be involved in the immune defense of intestinal tissues. This study provides a rich data base for in-depth study of the immune and metabolic responses of the honeybee midgut to infection by N. ceranae.