鉴定糖胰平治疗糖耐量受损的潜在治疗靶点：串联质谱标签标记的定量蛋白质组学分析

Identifying potential therapeutic targets of Tang-Yi-Ping for the treatment of impaired glucose tolerance: a tandem mass tag-labeled quantitative proteomic analysis.

作者信息

Li Jie, Bu Shuai, Zhou Honglei, Bi Siling, Xu Yunsheng

机构信息

College of the Second Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China.

Department of Endocrinology Medicine, the Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.

出版信息

Ann Transl Med. 2021 Oct;9(20):1532. doi: 10.21037/atm-21-4257.

DOI:10.21037/atm-21-4257

PMID:34790738

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8576661/

Abstract

BACKGROUND

This study uses the tandem mass tag (TMT)-labeled quantitative proteomic analysis to identify potential therapeutic protein targets of a Chinese prescription called Tang-Yi-Ping (TYP) for the treatment of impaired glucose tolerance (IGT) in rats.

METHODS

A total of 31 specific-pathogen free (SPF) male Wistar rats were used in our study. Ten were randomly selected as a control group, while 21 received a high-sugar and high-fat diet combined with an intraperitoneal injection of streptozotocin to establish IGT subjects. After eliminating 2 rats without successful modeling, 19 were randomly divided into a TYP group (n=9) and IGT model group (n=10). The TYP group was given a TYP decoction of 6.36 mg/kg-1/d-1. After 8 weeks of intervention, blood glucose-related indicators were measured, and cell morphology was observed by hematoxylin and eosin (HE) staining. TMT-labeled proteomic analysis was applied to detect the differentially expressed proteins (DEPs) in the pancreases of the three groups. The intersection of the DEPs in both the TYP group and IGT model group underwent Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses to identify the related biological functions and signal transduction pathways. Finally, western blot (WB) was used to verify the TMT proteomics results.

RESULTS

TYP can effectively reduce blood glucose and improve islet morphology in IGT rats. We identified a total of 16 potential therapeutic protein targets of TYP, 4 of which were upregulated, while 12 were downregulated, including Rbp4, Fam3b, Flot2, etc. [fold change (FC) >1.1, P<0.05]. The significant signal transduction pathways included arginine and proline metabolism, glyceride metabolism, glycerophospholipid metabolism, mTOR, Wnt, and insulin signaling pathways.

CONCLUSIONS

For anti-IGT therapy, we found TYP regulates 16 protein targets, multiple biological functions, and multiple signal transduction pathways. This study thus makes a significant contribution to identifying new potential therapeutic targets for treating IGT.

摘要

背景

本研究采用串联质谱标签（TMT）标记的定量蛋白质组学分析方法，以鉴定名为糖胰平（TYP）的中药复方治疗大鼠糖耐量受损（IGT）的潜在治疗性蛋白质靶点。

方法

本研究共使用31只无特定病原体（SPF）雄性Wistar大鼠。随机选取10只作为对照组，21只给予高糖高脂饮食并腹腔注射链脲佐菌素以建立IGT模型。在剔除2只建模未成功的大鼠后，将19只大鼠随机分为TYP组（n = 9）和IGT模型组（n = 10）。TYP组给予6.36 mg/kg-1/d-1的TYP水煎剂。干预8周后，检测血糖相关指标，并用苏木精-伊红（HE）染色观察细胞形态。应用TMT标记的蛋白质组学分析检测三组大鼠胰腺中差异表达蛋白（DEP）。对TYP组和IGT模型组中DEP的交集进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析，以鉴定相关生物学功能和信号转导通路。最后，采用蛋白质免疫印迹法（WB）验证TMT蛋白质组学结果。

结果

TYP可有效降低IGT大鼠的血糖并改善胰岛形态。我们共鉴定出16个TYP潜在治疗性蛋白质靶点，其中4个上调，12个下调，包括视黄醇结合蛋白4（Rbp4）、家族性肿瘤相关蛋白3b（Fam3b）、膜联蛋白2（Flot2）等[倍数变化（FC）>1.1，P<0.05]。显著的信号转导通路包括精氨酸和脯氨酸代谢、甘油酯代谢、甘油磷脂代谢、哺乳动物雷帕霉素靶蛋白（mTOR）、Wnt和胰岛素信号通路。