摘要
目的:通过定量观察补骨颗粒对大鼠膝关节软骨细胞蛋白质表达的影响,探讨补肾活血中药对软骨细胞的作用机制。方法:将软骨细胞经补骨颗粒含药血清传代培养后通过定量蛋白质组学串联质谱标签标记技术进行检测,使用Maxquant(v1.5.2.8)进行检索。选择UniProt-GOA数据库注释工具对差异蛋白进行基因功能聚类GO分析;采用KEGG在线服务工具KAAS对提交的蛋白进行注释,之后通过KEGGmapper将注释过的蛋白匹配入数据库相应的通路中进行检索分析;使用预测亚细胞定位的软件wolfpsort对所提交的蛋白进行亚细胞定位注释。结果:通过数据库检索,补骨颗粒含药血清及空白血清干预软骨细胞后共鉴定到5028个蛋白质,包含25423个肽段,其中4297个蛋白质包含定量信息。以1.3倍为变化阈值,t检验中P<0.05为标准,空白血清组中163个蛋白表达发生上调,188个蛋白表达发生下调;补骨颗粒含药血清干预后有164个蛋白表达发生上调,58个蛋白表达发生下调。其中,具有生物学信息的目标差异蛋白有腺苷酸转位酶1、热休克蛋白27、肌浆/内质网钙ATP酶1、钠/钾转运ATP酶、果糖二磷酸醛缩酶A、3-磷酸甘油醛脱氢酶、磷酸甘油酸变位酶1、β-烯醇化酶、L-乳酸脱氢酶、糖原合成酶、肉碱棕榈酰转移酶1。结论:补骨颗粒能够影响软骨细胞的腺苷酸转位酶1、热休克蛋白27、醛缩酶A、β-烯醇化酶及肉碱棕榈酰转移酶1等表达,对于揭示软骨细胞的凋亡机制具有重要意义。
Objective:By quantitatively observing the effects of Bugu Keli(补骨颗粒)on the expression of protein in knee joint chondrocyte of rats,to explore the mechanism of medicinals of nourishing kidney and activating the circulation of blood.Methods:Chondrocytes were subcultured in medicated serum with Bugu Keli and detected by quantitative proteomics tandem mass spectrometry tagging technology.Maxquant (v1.5.2.8)was used to retrieve.UniProt-GOA database annotation tool was selected to perform gene function clustering GO analysis of differentially expressed proteins;KEGG online service tool KAAS was used to annotate the submitted proteins,and then KEGG mapper was used to match the annotated proteins into the corresponding pathways in the database for retrieval and analysis.Wolfps was used to predict subcellular localization.Results:A total of 5028 proteins,including 25 423 peptide segments were identified after the intervention of medicated serum and blank serum on chondrocyte.Among them,4297 proteins contained quantitative information.With 1.3 times as the change threshold and P < 0.05 as the standard in the t test,163 protein expressions were upregulated and 188 protein expressions were down-regulated in the blank serum group.One hundred and sixty-four protein expressions were up-regulated and 58 protein expressions were down-regulated after the intervention of the drug-containing serum.Among them,the target differential proteins with biological information are adenylate translocatase 1,heat shock protein 27, Sarcoplasmic/endoplasmic reticulum calcium ATPase 1,sodium/potassium transporter ATPase,fructose diphosphate aldolase A,3-phosphate glyceraldehyde dehydrogenase,phosphoglycerol mutase 1,β-enolase,L-lactate dehydrogenase,glycogen synthase,and carnitine palmitoyltransferase 1.Conclusion:Bugu Keli can affect the expression of adenylate transposase 1,heat shock protein 27,aldolase A,β-enolase and carnitine palmitoyltransferase 1 in chondrocyte,which is of great significance in revealing the apoptotic mechanism
作者
余光书
林焱斌
许宏滨
李杰辉
张寿雄
YU Guang-shu;LIN Yan-bin;XU Hong-bin;LI Jie-hui;ZHANG Shou-xiong
出处
《风湿病与关节炎》
2019年第9期5-9,35,共6页
Rheumatism and Arthritis
基金
福建省自然科学基金资助项目(2016J01597)
福州市科技计划项目(2017-S-130-5)