基于差异蛋白组学的中医辨证治疗慢性萎缩性胃炎癌前病变机制研究

Gastric cancer is the most common malignant tumour in our country. The study on precancerous lesion of chronic atrophic gastritis(CAG) is an important content of secondary prevention of gastric cancer. The preliminary clinical observations and experimental studies we conducted approve that as precancerous lesion of CAG develops, TCM syndrome went through three stages: the first is blood stasis and heat toxin syndrome, second yin deficiency and heat excess syndrome and last qi and yin deficiency syndrome. We have explored the effect of compound Chinese herbs on the multiplication and apoptosis of gastric muscosal cells from the perspective of molecular biology by the municipal government grant. In order to get the complete picture of how precancerous lesion of CAG related DNA changes, for the first time ever, we intend to apply the technique of surface enhanced laser desorption and ionization time-of-light mass spectroscope (SELDI-TOF-MS) and establish the finger print, to reveal the differences of serum protein expression among the three syndromes mentioned above after being treated under the principal of promoting blood and detoxication, nourishing yin and clearing away heat, tonifying qi and yin pespectively, aiming at selecting the efficacy related protein as Chinese formula are usually multi-target. By analyzing TCM syndrome, endoscopic and pathological diagnosis, infection of HP in gastric mucosa as well as the relationship of the finger print of serum protein among different syndromes, the specific mass spectrum summit of different TCM syndromes and the target of the action can be confirmed, which helps to screen the protein as the target of Chinese medicine at molecular level and meanwhile lay the foundation for establishing proteomics data base of TCM syndromes and the protein as the target of the action of traditional chinese medicine.

胃癌是我国最常见的恶性肿瘤，对慢性萎缩性胃炎（CAG）癌前病变的研究是胃癌二级预防的重要内容。课题组经前期的临床观察及实验研究，认为其中医证侯变化过程为血瘀热毒、热毒伤阴、气阴两虚，并从分子生物学角度探讨了中药复方对胃黏膜细胞增殖凋亡的影响。为分析CAG癌前病变血清DNA改变的全貌，本研究拟利用蛋白质组学芯片-飞行时间质谱（SELDI-TOF-MS）技术，建立活血解毒法、养阴清热法、益气养阴法治疗上述三型CAG癌前病变患者差异血清蛋白质表达指纹图谱，针对中药多环节、多靶点作用特点，筛选出可用于疾病疗效机理分析的相关蛋白质。通过分析中医证候、胃镜肉眼诊断、病理学诊断、胃黏膜Hp感染以及各证型患者血清蛋白质谱之间的相互关联，明确中医证型和中药作用靶点特有的质谱峰,有利于从分子水平筛选中药的蛋白质作用靶点，为组建中医证型蛋白质组学信息库和中药作用靶点蛋白质组学信息库奠定基础。

胃癌是我国最常见的恶性肿瘤，对慢性萎缩性胃炎（CAG）癌前病变的研究是胃癌二级预防的重要内容。课题组经前期的临床观察及实验研究，认为其中医证侯变化过程为血瘀热毒、热毒伤阴、气阴两虚。按照中华中医药学会脾胃病分会的分型标准，64例CAG患者中医证型按出现频率由高到低依次是： 脾胃虚弱型>肝胃不和型>脾胃湿热型>胃阴不足型>胃络瘀阻型。应用SELDI-TOF-MS技术分析脾胃虚弱证、肝胃不和证、脾胃湿热证三个证型及健康对照各组间血清蛋白质谱的差异，从中筛选出各证型的差异血清蛋白质表达指纹图谱，寻找CAG癌前病变的差异蛋白及中医辨证的客观依据。CAG癌前病变各组与健康对照组之间的差异蛋白功能涉及到炎症反应、免疫反应、肿瘤细胞的迁移、再生及凋亡调控等。其中，β2GP与凝集素可能成为CAG癌前病变的标志性蛋白。胰蛋白酶抑制剂重链H4（ITIH4）可能是脾胃虚弱证区别于其他证型及健康对照组的标志蛋白。触珠蛋白可能是肝胃不和证区别于其他两个证型的标志蛋白。免疫球蛋白μ链c区、蛋白AMBP含量均降低，ITIH4均升高，此三者可能是脾胃湿热证区别于其他两个证型的标志蛋白。实验研究结果示CAG脾胃虚弱证型治疗前与健康对照组差异蛋白鉴定结果：分析得出46个差异蛋白点，上调的蛋白有33个，下调的蛋白有13个。给予升阳益胃汤治疗后与治疗前相比差异蛋白鉴定结果：分析得出10个差异蛋白点，上调的蛋白有4，下调的蛋白有6个。差异蛋白的生物过程主要涉及到免疫反应、炎症反应、信号转导、凋亡过程的负调节、趋化作用等。CAG脾胃虚弱证型与健康对照组之间的差异蛋白可能是其发病相关蛋白，外在证候的内在物质基础，对其进行验证对CAG脾胃虚弱证型的诊断有重要的指导作用。升阳益胃汤治疗CAG前后的蛋白质差异可能是中药作用治疗的靶点，及改善临床症状、阻止病变进一步发展的相关作用位点，有待进一步验证。