基于TMT蛋白质组学分析骨疏康胶囊治疗骨质疏松模型大鼠的机制  

Protective mechanism of Gushukang granule in a rat osteoporosis model based on TMT proteomic analysis

作　　者：林若慧 陈赛楠 叶云金 陈娟 谢丽华 黄景文 葛继荣 李生强 Lin Ruohui;Chen Sainan;Ye Yunjin;Chen Juan;Xie Lihua;Huang Jingwen;Ge Jirong;Li Shengqiang(Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China;Osteoporosis Syndrome Genomics Laboratory,Fujian Academy of Traditional Chinese Medicine,Fuzhou 350003,Fujian Province,China)

机构地区：[1]福建中医药大学,福建省福州市350122 [2]福建省中医药科学院骨质疏松证候基因组学研究室,福建省福州市350003

出　　处：《中国组织工程研究》2021年第32期5141-5147,共7页Chinese Journal of Tissue Engineering Research

基　　金：福建省科技厅省属公益类科研院所基本科研专项(2019R1003-1),项目负责人:李生强。

摘　　要：背景:骨疏康治疗骨质疏松症的具体机制及作用靶点尚未明确,需进一步深入研究。目的:探究骨疏康保护骨质疏松模型大鼠骨代谢的机制,对差异表达蛋白进行生物信息学分析。方法:36只雌性SD大鼠,去势法建立骨质疏松症模型大鼠24只,其中模型组12只,骨疏康组12只;假手术组12只为对照组。造模后,骨疏康组大鼠给予骨疏康灌胃[4.32 g/(kg·d)],1次/d;假手术组和模型组给予等体积生理盐水,1次/d。连续灌胃3个月后,测定大鼠胫骨骨密度;利用TMT-LC-MC/MC联用技术进行骨质疏松模型大鼠腰椎蛋白质组学分析,筛选出差异表达蛋白,对其进行GO、KEGG通路、蛋白互作分析,并构建蛋白质相互作用网络。实验方案经福建省中医药科学院动物实验伦理委员会批准。结果与结论:①筛选出骨疏康组/模型组之间32个上调和49个下调的差异表达蛋白;②GO富集分析表明,他们参与免疫球蛋白受体结合、血管内皮生长因子受体结合等生物过程及核酸模板转录、RNA生物合成等分子功能;③KEGG富集分析表明,骨疏康组与模型组的差异表达蛋白主要参与酪氨酸代谢、乙醇代谢、JAK-STAT等信号通路;④PPI分析表明,骨疏康组和模型组的共同差异蛋白中Mecp2、Aldh3b1、Ryr1、Myoz1位于蛋白质互作网络的节点且与骨代谢关系密切;⑤结果显示,骨疏康可能通过调控差异蛋白及酪氨酸代谢、JAK-STAT等重要信号通路参与保护骨质疏松模型大鼠的骨代谢过程。BACKGROUND:The specific mechanism and target of Gushukang in the treatment of osteoporosis are still unclear,which need further explorations.OBJECTIVE: To explore the protective mechanism of Gushukang on bone metabolism in osteoporotic rats and to analyze the differentially expressed proteins bybioinformatics.METHODS: Twenty-four osteoporosis model rats were established by ovariectomy, 12 in model group and 12 in Gushukang group. Another 12 rats withoutovariectomy were used as sham operation group. Gushukang capsule, 4.32 g/kg/d, was intragastrically administered once a day in the Gushukang group, andrats in the other two groups were given the same volume of normal saline. Treatment in each group lasted for 3 months. Bone mineral density of the tibia wasmeasured. The proteomic analysis of the lumbar vertebrae of osteoporotic rats was carried out by TMT-LC-MC/MC technique, and the differentially expressedproteins were screened, analyzed by gene ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein-protein interaction, therebyconstructing a protein-protein interaction network. An ethical approval was obtained from the Animal Ethics Committee of Fujian Academy of TraditionalChinese Medicine.RESULTS AND CONCLUSION: A total of 32 up-regulated and 49 down-regulated proteins were screened between Gushukang group and model group. Geneontology enrichment analysis showed that these differentially expressed proteins were involved in biological processes, such as immunoglobulin receptorbinding and vascular endothelial growth factor receptor binding, and molecular functions, such as nucleic acid template transcription and RNA biosynthesis.KEGG enrichment analysis showed that the differentially expressed proteins in the Gushukang group and model group were mainly involved in tyrosinemetabolism, ethanol metabolism, JAK-STAT and other signal pathways. Protein-protein interaction analysis showed that among the common differentialproteins in the Gushukang group and model group, Mecp2, Aldh3b1, Ryr1 and Myoz1

关 键 词：骨疏康 TMT技术 蛋白质组学 差异蛋白 骨质疏松模型大鼠 骨代谢 生物信息学 中医药 

分 类 号：R681[医药卫生—骨科学] R446[医药卫生—外科学]