基因芯片技术研究清胰汤对重症急性胰腺炎大鼠基因表达谱的影响  

Effect of Qingyi Decoction on Gene Expression Profiles of Severe Acute Pancreatitis Rats by Gene Chip Technique

作　　者：朱风尚[1] 朱国英 黄东平 沈晓莹[3] 杨长青[1] 郜恒骏[1,3] 

机构地区：[1]同济大学附属同济医院消化内科,上海200065 [2]上海普陀区人民医院院办公室,上海200060 [3]生物芯片上海国家工程研究中心,上海201203

出　　处：《中国中西医结合杂志》2014年第1期51-55,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：上海市普陀区科委科研基金(No.PTKW08-C03)

摘　　要：目的研究清胰汤(Qingyi Decoction,QYD)对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠胰腺组织基因表达谱的影响。方法 60只SD大鼠按随机数字表法分为假手术组、SAP组和QYD组,每组20只。4%牛磺胆酸钠胰胆管逆行注射复制SAP模型,QYD组3次灌胃治疗(0.75 mL/100 g)。Illumina全基因组表达谱芯片分析胰腺表达谱变化,实时定量PCR和Western blot验证芯片部分基因热休克蛋白a8(heat shock proteins a8,Hspa8)、热休克蛋白b1(heat shock proteins b1,Hspb1)及其蛋白表达。结果与SAP组比较,QYD组筛选出575个差异基因,其中上调92个,下调483个。基因本体论(Gene Ontology,GO)分析涉及到转录调节因子活性负调节、氧化还原酶类活性、酶抑制剂活性等。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库分析其主要涉及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路、NOD样受体(NOD like receptors,NLR)信号通路、细胞周期、代谢通路、氧化还原酶类活性等。PCR和Western blot验证芯片中Hspa8和Hspb1相对mRNA表达分别升高(13.24±1.22)倍和(7.55±1.09)倍(P<0.01)。结论 QYD有效治疗实验性SAP机制涉及到MAPK信号通路、NLR信号通路、细胞周期、代谢通路、氧化还原酶类活性等。Objective To investigate the effect of Qingyi Decoction （QYD） on pancreatic gene expression profiles in rats with severe acute pancreatitis （SAP）. Methods Totally 60 Sprague-Dawley （SD） rats were randomly divided into the sham-operation group （SO group）, the SAP group, and the QYD group, 20 in each group. SAP model was replicated by the pancreatic duct retrograde injection with 4% sodium taurocholate. Rats in the QYD group was intragastrically intervened by QYD （0.75 mL/100 g） for 3 times. Pancreatic RNA expression was analyzed using Illumina whole genome expression profiles. Changes of mRNA and protein in specific genes Eheat shock proteins a8 （Hspa8） and heat shock proteins bl （Hspbl） ~ were verified by real-time quantitative PCR and Western blot analysis. Results Compared with the SAP group, 575 differential genes were screened in the QYD group, including 92 up-regulated genes and 483 down-regulated genes. Gene Ontology （GO） categories indicated the genes are asso- ciated with negative regulation of transcription regulator activity, oxidoreductase activity and enzyme inhibitor activity. Effects of QYD on the SAP rats were majorly related to mitogen-activated protein kinase （MAPK）, NOD like receptors （NLR） receptor-like signaling pathway, cell cycle, metabolic pathways, oxidoreductase activity. Protein and mRNA changes of Hspa8 and Hspbl in microarray were verified [ rel- ative mRNA expression for Hspa8 and Hspbl was increased by （13.24 ±1.22） times and （7.55±1.09） times respectively, P 〈0.01 ]. Conclusion QYD was effective in treating experimental SAP involved the MAPK and NLR signaling pathways, cell cycle, metabolic pathways, and oxide reductase activities.

关 键 词：重症急性胰腺炎 清胰汤 基因表达谱 聚合酶链式反应 蛋白质印迹法 

分 类 号：R285.5[医药卫生—中药学]