基于iTRAQ技术分析脾气虚证大鼠回肠蛋白质的差异  

Analysis of Difference of Ileal Protein in Rats with Spleen-qi Deficiency Syndrome Based on Isobaric Tags for Relative and Absolute Quantitation Technology

作　　者：杨关林 宋囡 贾连群 王德山 单德红 张哲 马艺鑫 张妮 

机构地区：[1]辽宁中医药大学中医脏象理论及应用教育部重点实验室 [2]辽宁中医药大学基础医学院 [3]辽宁中医药大学附属医院

出　　处：《中医杂志》2017年第24期2124-2127,共4页Journal of Traditional Chinese Medicine

基　　金：国家重点基础研究发展计划("973"计划)(2013CB531704);国家自然科学基金(81202834);辽宁省高等学校优秀人才支持计划(LR2015041)

摘　　要：目的探讨脾气虚证分子生物学特征。方法 12只SD大鼠随机分为正常组和脾气虚组各6只。脾气虚组采用饮食不节加力竭游泳建立脾气虚证模型。分离提取回肠总蛋白,用相对和绝对定量同位素标记(i TRAQ)法检测分析差异表达的蛋白质种类及两组表达比值,用Western blot法验证差异蛋白小肠脂肪酸结合蛋白(I-FABP)、肝脂肪酸结合蛋白(L-FABP)、尼曼匹克C1型类似蛋白(NPC1L1)的表达量。结果i TRAQ技术分析发现两组大鼠脂肪消化及吸收相关通路蛋白存在差异,与正常组比较,脾气虚组I-FABP、微粒体甘油三酸酯转移蛋白(MTTP)、L-FABP、NPC1L1、载脂蛋白B(ApoB)、载脂蛋白A4(ApoA4)及单酰甘油酰基转移酶(Mogat 2)7个蛋白均出现了上调(变化倍数﹥1.2)。Western blot法验证差异蛋白IFABP、L-FABP、NPC1L1变化趋势与蛋白质组学结果相一致。结论脾气虚证大鼠回肠组织中I-FABP、MTTP、L-FABP、NPC1L1、ApoB、ApoA4及MGAT7蛋白表达上调。Objective To explore the molecular biological characteristics of spleen-qi deficiency syndrome. Methods Twelve Sprague Dawley rats were randomized into a normal group and a spleen-qi deficiency group （n = 6 in each）. Rats in the spleen-qi deficiency group were made spleen-qi deficiency models by improper diet and exhaus- tive swimming. Ileal total protein was separated and extracted. The differential expression proteins and the expression ratio in both groups were detected and analyzed with isobaric tags for relative and absolute quantitation （iTRAQ） method. The expressions of differential expression proteins, such as small intestinal fatty acid binding protein （I- FABP） , liver fatty acid binding protein （L-FABP） and Niemann-Pick type C1 Like 1 （NPC1L!） , were verified by Western blot method. Results iTRAQ technology analysis showed that there were difference in fatty digestion and absorption related pathway proteins in rats between the two groups. Compared with those in the normal group, I- FABP, microsomal triglyceride transfer protein （MTYP）, L-FABP, NPC1L1, apolipoprotein B （ApoB）, apolipopro- tein A4 （ApoA4） and monoacylglycerol acyl transferase （Mogat 2 ） in the spleen-qi deficiency group were up- regulated （fold change more than 1. 2）. It was verified by Western blot method that the change trend of differential proteins I-FABP, L-FABP and NPC1L1 was in accord with the result of proteome. Conclusion The expressions of I-FABP, MTTP, L-FABP, NPC1L1, ApoB, ApoA4 and MGAT7 proteins in ileal tissue of rats with spleen-qi deficiency syndrome are up-regulated.

关 键 词：脾气虚证 回肠 蛋白质组学 脂肪酸 

分 类 号：R228[医药卫生—中医基础理论] R-332[医药卫生—中医学]