基于蛋白组学和NCBI数据库探究反流性食管炎及细胞焦亡调控基因  

Regulatory genes of reflux esophagitis and cell pyroptosis based on proteomics and NCBI database

作　　者：刘思雨 唐艳萍[2] 刘磊 李培彩[2] 刘茜 刘琰 杨磊[3] Liu Siyu;Tang Yanping;Liu Lei;Li Peicai;Liu Xi;Liu Yan;Yang Lei(Institute of Graduate,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Department of Gastroenterology,Tianjin Nankai Hospital,Tianjin 300100,China;Institute of Integrated Chinese and Western Medicine Acute Abdomen,Tianjin Nankai Hospital,Tianjin 300100,China)

机构地区：[1]天津中医药大学中西医结合临床,301617 [2]天津市南开医院消化内科二,300100 [3]天津市南开医院天津市中西医结合急腹症研究所,300100

出　　处：《中华实验外科杂志》2022年第4期676-678,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81573737、82074213)。

摘　　要：目的验证经典细胞焦亡途径在反流性食管炎(RE)发病中的作用并筛选出部分介导RE发病并调控细胞焦亡的基因。方法建立RE大鼠模型,实验分为正常对照组和模型组。苏木精-伊红(HE)染色法观察食管组织病理学变化,透射电镜观察食管黏膜上皮细胞形态,免疫组织化学染色法检测食管组织中NOD样受体蛋白3(NLRP3)、半胱天冬酶1(Caspase-1,CASP1)表达,蛋白免疫印迹法检测食管组织中Gasdermin D(GSDMD)表达,采用方差分析进行统计分析。取大鼠食管下段组织进行蛋白组学分析,基于蛋白组学和NCBI数据库对RE调控基因及细胞焦亡相关基因进行比较,筛选出部分介导RE发病并调控细胞焦亡的基因。结果RE大鼠食管组织病理评分增高,肿大变形及破裂穿孔的食管黏膜上皮细胞数目增多,NLRP3、Caspase-1、GSDMD表达显著增加。蛋白组学分析获取的差异蛋白及其编码基因与NCBI中GENE数据库所获取的焦亡相关基因进行交叉分析,获取到9个重叠基因,其中6个表达上调,分别为半胱天冬酶8(Caspase-8,CASP8)、DExH-box解旋酶9(DHX9)、半胱天冬酶3(Caspase-3,CASP3)、间隙连接蛋白α1(GJA1)、组织蛋白酶B(CTSB)、蛋白酶体20S亚基beta 8(PSMB8)。3个表达下调,分别为APAF1相互作用蛋白(APIP)、免疫球蛋白(BSG)、脂联素(ADIPOQ)。结论细胞焦亡在RE的发病机制中发挥重要作用,CASP8、DHX9、CASP3、GJA1、CTSB、PSMB8、APIP、BSG、ADIPOQ这9个重叠基因可能通过调控细胞焦亡参与RE的发生发展。Objective To verify the role of classical cell pyroptosis pathway in the pathogenesis of reflux esophagitis(RE)and to screen some genes that mediate the pathogenesis of RE and regulate cell pyroptosis.Methods In this study,RE models of rats were established,and the rats were divided into normal control group and model group.The histopathological changes of esophagus were observed by haematoxylin and eosin(HE)staining.The morphological changes of esophageal epithelial cells were observed by transmission electron microscope.The expression levels of NLR family pyrin domain containing 3(NLRP3)and Caspase-1(CASP1)in esophageal tissues were detected by immunohistochemical staining,and the expression of Gasdermin D(GSDMD)in esophageal tissues was detected by Western blotting.Data were analyzed by ANOVA.The lower esophageal tissues of rats were taken for proteomic analysis.Based on proteomics and NCBI database,RE regulatory genes and genes related to cell pyroptosis were compared,and some genes that mediate RE pathogenesis and regulate cell scorch death were screened out.Results In RE rats,the histopathological score of esophagus increased,the number of esophageal epithelial cells with swelling,deformation,rupture and perforation increased,and the expression levels of NLRP3,Caspase-1 and GSDMD increased significantly.The differential proteins and their coding genes obtained from proteomics analysis were cross-analyzed with the genes related to cell pyroptosis obtained from GENE database in NCBI,and 9 overlapping genes were obtained,among which 6 genes were up-regulated,namely Caspase-8(CASP8),DExH-box helicase 9(DHX9),Caspase-3(CASP3),gap junction protein alpha 1(GJA1),cathepsin B(CTSB)and proteasome 20S subunit beta 8(PSMB8),and 3 genes were down-regulated respectively,namely APAF1 interacting protein(APIP),basigin(BSG)and Adiponectin(ADIPOQ).Conclusion Cell pyroptosis plays an important role in the pathogenesis of RE.A total of 9 overlapping genes,including CASP8,DHX9,CASP3,GJA1,CTSB,PSMB8,APIP,BSG and ADIPOQ,may parti

关 键 词：反流性食管炎 细胞焦亡 蛋白组学 

分 类 号：R571[医药卫生—消化系统]