清胰颗粒干预重症急性胰腺炎后胰腺蛋白组学表达差异性研究

Differentially Expressed Proteins of Severe Acute Pancreatitis Intervened by Qingyi Granule

目的观察清胰颗粒(Qingyi Granules,QYG)对牛黄胆酸钠(sodium tauro cholate,STC)诱导大鼠重症急性胰腺炎(severe acute pancreatitis,SAP)胰腺组织中总蛋白表达变化的影响。方法 36只SD大鼠经胰胆管逆行注射5%STC诱导SAP,随机分为SAP组和QYG治疗组(QYG组),每组18只。建模成功后,QYG组每12h1次QYG对水(W∶W=1∶1)灌胃[1mL/(100g·只)],SAP组以生理盐水替代,共给药4次。提取两组大鼠胰腺组织总蛋白行双向电泳(two-dimensional electrophoresis,2-DE)、荧光染色和图谱分析,选择48h两组间差异表达超过4倍的蛋白质点进行MALDI-TOF/TOF质谱分析和生物信息学分析,观察两组蛋白质点的差异。结果 5%STC诱导出SAP大鼠模型,胰腺组织有典型病理学改变。经凝胶图像处理软件分析胰腺组织蛋白组学变化:48h时两组有22个差异点,用药后5个点蛋白上调,17个点下调。48h时间点比较两组表达差异超过4倍且稳定的蛋白质斑点共有9个,质谱分析和数据库检索共鉴定出7种蛋白质,分别是丝氨酸蛋白酶抑制剂b1a(39kb)、丝氨酸蛋白酶抑制剂b1a(43kb)、硫氧还蛋白过氧化物酶-Ⅳ(PrxⅣ)、糜蛋白酶样蛋白(Clps)、γ-肌动蛋白(Actg1)、谷氨酰-脯氨酰tRNA合成酶(Eprs)、短链羟酰辅酶A脱氢酶(Hadhsc)。从功能分析,这些蛋白与SAP胰腺病理损伤过程中信号传导相关。结论比较蛋白质组学能较好地反映QYG对SAP大鼠胰腺组织中差异表达蛋白的影响,研究差异表达蛋白有可能为QYG治疗SAP提供新的理论基础和分子靶点。

Objective To observe the effects of Qingyi Granule （QYG）on the changes of total protein expressions in the pancreatic tissue of rats with severe acute pancreatitis （SAP）induced by sodium taurocholate （STC）. Methods SAP was induced by retrograded injecting 5% STC from the gut-pancreatic duct in 36 Sprague-Dawley （SD）rats. Then they were randomly divided into the SAP group and the QYG treatment group （abbreviated as the QYG group）, 18 in each group. After successful modeling, rats in the QYG group were administered with QYG water solution （W：W=1：1）once with an interval of 12 h （1 mL/100 g）, while rats in the SAP group were administered with normal saline. The medication was performed four times. The total proteins were extracted from the pancreatic tissue of all rats to perform two-dimensional electrophoresis, fluorescent staining, and atlas analysis. The protein dots with differential expressions more than four times between each other in 48 h gel pictures were chosen and used for MALDI-TOF/TOF mass chromatographic analysis and biological information analysis. Results The 5% STC induced SAP model rats had typical pathological changes in the pancreatic tissue. The proteomics changes of the pancreatic tissue were analyzed by gel image manipulation software. Twenty two disparate points were detected between two groups at 48 h, 5 points of the protein were up-regulated and 17 points were down-regulated of the total after QYG intervention. Nine protein spots expressed differently more than 4 times and stably at 48 h, 7 kinds of proteins have been identified by mass chromatographic analysis and Data Base Retrieval, and they were Serpinbla 39 kDa protein, Serpinbla 43 kDa protein, Prdx4 Prx IV, Clps, h,-actin （Actgl）, Eprs and Hadhsc. Those proteins were involved in signal transmit during the process of SAP pancreas pathological injury analyzed from their functions. Conclusions Proteomics can well reflect the effects of QYG on differential expression proteins in the pancreatic tissue of rats with SAP. Studying differential expression proteins may provide a new theoretical basis and molecule target for QYG treating SAP.