成年自发性高血压大鼠肠系膜动脉蛋白组学分析

Proteomics analysis of mesenteric artery in adult spontaneously hypertensive rats

目的应用蛋白组学研究成年自发性高血压大鼠(SHR)肠系膜动脉血管重塑过程中的蛋白表达情况。方法20周龄雄性Wistar-Kyoto(WKY)大鼠和SHR各15只,各随机选取6只进行三级肠系膜动脉的形态学检测,剩余各9只进行肠系膜三级动脉的蛋白组学分析。采用液相色谱串联质谱进行蛋白鉴定和定量,通过生物信息学分析差异表达蛋白。结果与WKY大鼠相比,SHR肠系膜动脉的管壁厚度/管腔半径(0.21±0.02比0.17±0.02,t=3.286,P=0.008)及管壁面积/管腔面积(0.46±0.06比0.37±0.04,t=3.272,P=0.009)明显增高。两种成年大鼠肠系膜动脉中有277个差异表达蛋白,SHR中143个蛋白表达上调,134个蛋白表达下调。化学突触传递、肽抗原结合和质膜分别是最显著富集的基因本体(GO)生物学过程、分子功能和细胞成分条目。细胞色素P450对异源物质的代谢是最显著富集的京都基因和基因组百科全书(KEGG)通路。在蛋白相互作用网络分析中,核糖体蛋白L26(RPL26)、还原型烟酰胺腺嘌呤二核苷酸(磷酸)醌脱氢酶1(NQO1)、钙黏蛋白2(CDH2)、谷胱甘肽S-转移酶mu4(GSTM4)、胎盘碱性磷酸酶(ALPP)、von Willebrand因子(vWF)、丝氨酸家族C成员1(SERPINC1)与其他差异表达蛋白关联度最高。结论蛋白组学分析显示,在成年SHR肠系膜动脉血管重塑过程中,化学突触传递和细胞色素P450对异源物质的代谢等相关蛋白表达发生了变化,这可能为高血压阻力血管重塑机制的深入研究提供新思路。

Objective To investigate the changes of protein expression in the mesenteric artery during vascular remodeling in adult spontaneously hypertensive rats(SHR)by using proteomics analysis.Methods Fifteen male Wistar-Kyoto(WKY)rats and 15 SHR at the age of 20 weeks were used.Six of them were randomly selected to observe the morphology of the third-order branches of mesenteric arteries,and the remaining nine of them were subjected to proteomic analysis of the third-order branches of mesenteric arteries.Liquid chromatography/tandem mass spectrometry analysis was used to identify and quantify the expression of protein,and bioinformatics analysis was applied to analyze the differentially expressed proteins.Results Compared with WKY rats,the wall thickness to lumen radius(0.21±0.02 us 0.17±0.02,t=3.286,P=0.008)and wall area to lumen area(0.46±0.06 vs 0.37±0.04,t=3.272,P=0.009)of mesenteric arteries in SHR were significantly increased.There were 277 differentially expressed proteins in mensenteric artery between two kinds of adult rats,including 143 upregulated proteins and 134 downregulated proteins in SHR.Chemical synaptic transmission,peptide antigen binding and plasma membrane were the most significantly enriched Gene Ontology(GO)biological process term,molecular function term and cellular component term,respectively.Metabolism of xenobiotics by cytochrome P450 was the most markedly enriched Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.In the protein-protein interaction network,ribosomal protein L26(RPL26),reduced nicotinamide adenine dinucleotide phosphate quinone dehydrogenase 1(NQO1),cadherin 2(CDH2),glutathione S-transferase mu 4(GSTM4),alkaline phosphatase,placental(ALPP),von Willebrand factor(vWF),serpin family C member 1(SERPINC1)had the highest correlation with other differentially expressed proteins.Conclusion Proteomic analysis shows the expression change of protein related to chemical synaptic transmission and metabolism of heterologous substances by cytochrome P450 during mesenteric artery vascular remodeling in adult SHR,which provides insight into the mechanism of hypertensive resistance vascular remodeling.