文献类型：期刊文献

中文题名：基于蛋白质组学研究小儿开胃增食合剂对小儿厌食症模型大鼠空肠蛋白表达的影响及机制

英文题名：Effect of Xiaoer-Kaiwei-Zengshi mixture on jejunal protein expression in rats with infantile anorexia and its therapeutic mechanism based on proteomics

作者：呼荟茹[1];王小荣[1];吴丽萍[2];史正刚[1]

第一作者：呼荟茹

机构：[1]甘肃中医药大学,甘肃兰州730030;[2]甘肃中医药大学附属医院,甘肃兰州730030

第一机构：甘肃中医药大学

年份：2022

卷号：38

期号：7

起止页码：1253

中文期刊名：中国病理生理杂志

外文期刊名：Chinese Journal of Pathophysiology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金资助项目(No.81360556)。

语种：中文

中文关键词：小儿开胃增食合剂;厌食症;蛋白质组学

外文关键词：Xiaoer-Kaiwei-Zengshi mixture;Infantile anorexia;Proteomics

摘要：目的:基于蛋白组学阐明小儿开胃增食合剂(Xiaoer-Kaiwei-Zengshi mixture,XKZM)治疗小儿厌食症(infantile anorexia,IA)大鼠的作用机制,鉴定其关键蛋白,并探索其可能的通路。方法:利用特制饲料诱导建立SD大鼠IA模型,随后将造模成功的大鼠随机分为模型组(model组)、西药多潘立酮(domperidone,DPD)对照组(DPD组)、低剂量XKZM(low-dose XKZM,LXKZM)组、中剂量XKZM(medium-dose XKZM,MXKZM)组和高剂量XKZM(high-dose XKZM,HXKZM)组,另设空白对照组(control组),每组6只。对造模期间和干预期间大鼠的进食量和体重进行测量;HE染色观察大鼠空肠组织病理变化;基于串联质量标签(tandem mass tag,TMT)定量蛋白质组学技术对大鼠空肠组织中的差异蛋白进行检测,对差异蛋白进行GO(Gene Ontology)分析、KEGG(Kyoto Encyclopedia of Genes and Genomes)通路分析及相互作用分析,Western blot实验验证XKZM调控的差异蛋白表达,筛选XKZM的核心靶点。结果:与control组相比,model组大鼠体重和进食量显著降低(P<0.05),空肠组织出现结构损伤和炎症反应;给予IA大鼠XKZM干预后,LXKZM、MXKZM和HXKZM组大鼠体重和进食量较model组均显著增加(P<0.05),胃组织的病理变化明显改善,并均呈剂量依赖性。蛋白组学结果显示,本次研究中共定量了4 116种蛋白,与control组比较,model组空肠组织中检测到32个差异蛋白(上调和下调蛋白数分别为20和12);与model组比较,XKZM组检测到71个差异蛋白(上调和下调蛋白数分别为29和42);GO分析表明这些差异蛋白主要被富集到细胞过程、ATP结合、抗氧化活性、蛋白结合和信号转导等过程;KEGG通路分析表明差异蛋白被富集到了糖酵解和葡萄糖代谢合成、生物合成、胰岛素相关信号等信号通路。蛋白相互作用分析结果表明,Nqo1[NAD(P)H quinone dehydrogenase 1]、Gsta4(glutathione S-transferase alpha 4)、ApoE(apolipoprotein E)、Gpx1(glutathione peroxidase 1)、Gpd2(glycerol-3-phosphate dehydrogenase 2)等蛋白作为枢纽与其它蛋白发生相互作用。Western blot结果表明,与control组比较,model组大鼠空肠组织中SULT1C2(sulfotransferase family 1C member 2)和CRABP2(cellular retinoic acid-binding protein 2)蛋白表达减少(P<0.05),ENO3(enolase 3)和KRT5(keratin 5)蛋白表达增加(P<0.05);与model组比较,XKZM组SULT1C2和CRABP2蛋白表达增加(P<0.05),ENO3和KRT5蛋白表达减少(P<0.05)。这些差异蛋白的表达变化趋势同蛋白组学结果一致。结论:XKZM可提高IA大鼠空肠组织中血脂代谢相关因子表达,进一步可能通过促进血脂代谢,影响与之相关的糖酵解和葡萄糖代谢合成、氨基酸的生物合成、碳代谢、ATP生成等信号通路,达到治疗IA的效果。
AIM:To elucidate the mechanism of Xiaoer-Kaiwei-Zengshi mixture(XKZM)in the treatment of infantile anorexia(IA)in rats based on proteomics.METHODS:The IA model in SD rats was induced by special feed,and then the rats were randomly divided into control group,model group,domperidone(DPD)group,low-dose XKZM(LXKZM)group,medium-dose XKZM(MXKZM)group and high-dose XKZM(HXKZM)group,with 6 rats in each group. The food intake and body weight of rats during modeling and intervention were measured. The pathological changes of rat jejunal tissues were observed by HE staining. The differential proteins in rat jejunum were detected based on tandem mass tag(TMT)quantitative proteomic technique,and the Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways and interactions of the differential proteins were analyzed. Western blot was used to verify the differential protein expression regulated by XKZM,and screen the core target of XKZM.RESULTS:Compared with control group,the body weight and food intake were decreased significantly in model group(P<0. 05). Significant pathological injury and inflammatory response were observed in the jejunal tissue of the rats in model group. Compared with model group,the weight and food intake in anorexic rats were significantly increased in XKZM groups(P<0. 05),and the pathological changes of gastric tissue were significantly improved in a dose-dependent manner. Proteomic results showed that a total of 4 116 proteins were quantified in this study. Compared with control group,32 differential proteins were found in the jejunum tissue of the rats in model group,and the numbers of up-regulated and down-regulated proteins were 20 and12,respectively. Compared with model group,71 differential proteins were found in XKZM group,including 29 up-regulated and 42 down regulated proteins. The results of GO enrichment analysis showed that these differential proteins were mainly enriched in cellular processes,ATP binding,antioxidant activity,protein binding and signal transduction. The results of KEGG pathway analysis showed that the differential proteins were enriched in glycolysis and glucose metabolism synthesis,biosynthesis,insulin-related signals and other signal pathways. The results of protein interaction analysis showed that Nqo1[NAD(P)H quinone dehydrogenase 1],Gsta4(glutathione S-transferase alpha 4),ApoE(apolipoprotein E),Gpx1(glutathione peroxidase 1)and Gpd2(glycerol-3-phosphate dehydrogenase 2)interacted with other proteins as hubs. The results of Western blot showed that compared with control group,the protein expression levels of SULT1C2(sulfotransferase family 1C member 2)and CRABP2(cellular retinoic acid-binding protein 2)were decreased in model group(P<0. 05),while the expression levels of ENO3(enolase 3)and KRT5(keratin 5)proteins were increased(P<0. 05). Compared with model group,the expression levels of SULT1C2 and CRABP2 proteins were increased in XKZM groups(P<0. 05),while the expression levels of ENO3 and KRT5 proteins were decreased(P<0. 05). The expression trend of these differential proteins was consistent with the results of proteomics.CONCLUSION:Treatment with XKZM promotes the expression of lipid metabolism-related proteins in the jejunum of IA model rats.