机械牵张力下大鼠小梁网细胞的差异蛋白组学及生物信息学分析

Differentially proteomics and bioinformatics analysis of rat trabecular meshwork cells under mechanical strain

目的研究机械牵张作用对大鼠小梁网细胞蛋白组学的表达影响。方法实验对象选用对数生长期的大鼠小梁网细胞。采用FX-5000牵张应力加载系统拉伸细胞,提取细胞全蛋白后进行基于鸟枪法的质谱测试和蛋白质的鉴定。运用微阵列显著性分析(significance analysis of microarray,SAM)方法分析实验组和对照组的蛋白组学差异;对差异蛋白进行基于Gene Ontology(GO)的功能注释、富集和Kyoto Encyclopedia of Genes and Genomes(KEGG)通路富集并筛选出显著富集的蛋白。结果 SAM法筛选出差异蛋白共计57种,均表达上调,分布在细胞各个区域,参与细胞发育、免疫调节、刺激反应、代谢、细胞黏附等过程。其中24种显著富集在9个隶属生物学过程的GO term中,进一步KEGG通路富集分析显示热休克蛋白和核糖体蛋白显著富集在内质网蛋白加工通路和核糖体通路中。结论机械牵张对大鼠小梁网细胞的蛋白质表达影响区域广,过程复杂。富集分析说明热休克蛋白和核糖体蛋白在牵张刺激下的应答保护和眼压调控中起到了重要作用。

Objective To study differential proteomics analysis of rat trabecular meshwork cells under mechanical stimulation.Methods The rat trabecular meshwork cells in logarithmic growth period were selected.Mechanical stretching was loaded by FX-5000 tension system.The total proteins were extracted and identified by shotgun-based mass spectrometry test and analysis.Differential expressed proteins between sample group and control group were analyzed by significance analysis of microarray(SAM)method.Gene Ontology(G O)annotation,GO enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were used to gain significantly enriched differential proteins.Results A total of 57 differential proteins were screened by SAM m ethod,all of which were up-regulated and distributed in various regions of the ce ll,and were involved in the processes of cell development,immunomodulation,stimulus response,metabolism and cell adhesion.Twenty-four of them were significantly enriched in 9 biological process CO terms and 2 KECC pathways,mainly assigned to heat shock proteins and ribosome proteins.Conclusions The effect of mechanical strain on protein expression of rat trabecular meshwork cells was wide and complex.The enrichment analysis showed that heat shock proteins and ribosome proteins played an important role in the response protection and intraocular pressure regulation under mechanical strain.