消痰散结方对裸鼠人胃癌MKN-45皮下移植瘤模型血清蛋白质组表达的影响

摘要

目的 观察消痰散结方对裸鼠人胃癌MKN-45皮下移植瘤模型血清蛋白质组表达的影响.方法 30只BALB/c-nu/nu裸鼠随机分为正常组、模型组、干预组,采用瘤块接种方法造模.正常组自由饮食,模型组、干预组分别给予生理盐水、消痰散结方灌胃.运用蛋白质组学的双向凝胶电泳技术,观察3组之间的血清蛋白质组表达差异,筛选出与消痰散结方作用相关的差异表达蛋白质,并对其进行质谱鉴定,确认差异表达蛋白质的身份.结果 去除血清中高丰度蛋白质后获得分辨率高、重复性稳定性好的血清2-DE图谱.与正常组比较,模型组中找到25个差异表达蛋白点,其中12个表达上调,13个表达下调.与模型组比较,干预组中找到19个差异表达蛋白点,其中14个表达上调,5个表达下调.3组间比较共找到差异表达蛋白点9个,与正常组比较,在模型组中表达上调、在干预组表达回调至正常组水平的3个,最终鉴定为结合珠蛋白、泛素蛋白连接酶、组蛋白甲基转移酶;与正常组比较,在模型组中表达下调、在干预组表达回复至正常组水平的有6个,其中3个鉴定为载脂蛋白A1、过氧化物酶1、超氧化物歧化酶.结论 消痰散结方对载脂蛋白A1、过氧化物酶1等功能蛋白表达的综合性调节可能是其发挥抗胃癌作用的重要机制.%Objective To observe the serum protein spectrum changes of MKN-45 gastric cancer cell bearing nude mice intervened by Xiaotan Sanjie Recipe on the platform of proteomics. Methods Thirty BALB/c-nu/nu mice were randomly divided into normal group, model group and intervention group. MKN45 gastric cancer cells were subcutaneously implanted into immune-incompetent nude mice in model group and intervention group to establish tumor bearing model. The control group was fed freely. The model group and the intervention group was separately administrated with saline and Xiaotan Sanjie Recipe, which were delivered by intragastric administration. Observe and analyze the differently expressed protein spots of the serum samples by checking two-dimensional gel electrophoresis maps of the three groups. The most differentially expressed proteins were digested, analyzed with MALDI-TOF/TOF-MS mass spectrograph and searched from protein database. Results The two-dimensional gel electrophoresis maps obtained good reproducibility and stability after depletion the high abundance proteins. By comparison to control group, 25 differently expressed protein spots were obtained in model group, 12 of which were promoted and 13 depressed. By comparison to model group, 19 different protein spots were obtained in intervention group, fourteen of which were promoted and 5 depressed. There were 9 proteins which had significant difference between intervention group and model group, three of which were promoted in model group compared with control group, but depressed to the normal level in the intervention group, identified as haptoglobin, ubiquity protein ligase, histone methyltransferase. Six proteins were depressed in model group compared with control group, but promoted to the normal level in the intervention group, three of them were identified as APOA1, peroxiredoxin1, superoxide dismutase. Conclusion Xiaotan Sanjie Recipe maybe plays an anti-tumor effects through comprehensively regulating the functional proteins such as haptoglobin, ubiquity protein ligase, et al.