基础医学与临床2024，Vol.44Issue(5)：690-698,9.DOI:10.16352/j.issn.1001-6325.2024.05.0690
应用微量蛋白质定量分析评估临床样本炎性反应状态
Quantitative analysis of trace-level proteins for assessing inflammation through clinical sampling
摘要
Abstract
Objective To compare the effectiveness between qPCR and trace-level proteins quantitative analysis in detecting the expression of inflammation-associated factors caspase-1 and IL-1β in peripheral blood monocytes from breast cancer patients.Methods Pre-and post-operation peripheral blood samples were collected from 10 breast cancer patients.Monocytes were subsequently isolated and activated by incubation with lipopolysaccharide.The mRNA and protein expression level of caspase-1 and IL-1 β in monocytes were examined using qPCR and trace-level proteins quantitative analysis,respectively,and the data were normalized by β-actin.The differences between the two detection methods were compared and evaluated for strengthens and weakness.Results The vast majority of patients showed a notable increase in the number of peripheral blood monocytes after surgical operation.At the mRNA level,compared to preoperative levels,postoperative caspase-1 and IL-1 β were both increased in 50％(5/10)and 40％(4/10)of patients,respectively,while the rest showed either a decrease(40％and 50％,re-spectively)or no significant change(both 10％).In contrast,the results of trace-level proteins quantitative analysis revealed that 60％(6/10)of patients exhibited increased level of postoperative caspase-1 and IL-1 β pro-teins,while the remaining showed a decrease(30％and 20％,respectively)or no significant change(10％and 20％,respectively).Additionally,50％of patients displayed a significant increase in caspase-1 and IL-1 β active cleavage bands by electrophoresis postoperatively.Notably,inconsistent changes in caspase-1 and IL-1β mRNA and protein level after surgery were observed in 30％and 40％of patients respectively.Conclusions The trace-level proteins quantitative analysis is recommended due to its advantages including minimal sample amount requirement,high sensitivity,good repeatability and relatively simple operative procedure.Therefore,it is particularly suitable for assessing the inflammatory status of of patients through clinical sampling of peripheral blood.关键词
炎性相关因子/caspase-1/IL-1β/微量蛋白质定量分析系统/实时荧光定量PCR(qPCR)Key words
inflammation-associated factors/caspase-1/IL-1β/trace-level proteins quantitative analysis/real-time quantitative pol-ymerase chain reaction(qPCR)分类
医药卫生引用本文复制引用
叶雨昕,李慧娴,阎涛,张赟,孙玉琳..应用微量蛋白质定量分析评估临床样本炎性反应状态[J].基础医学与临床,2024,44(5):690-698,9.基金项目
国家自然科学基金(82073327) （82073327）
中国医学科学院医学与健康科技创新工程经费(2021-I2M-1-066) （2021-I2M-1-066）
国家重点研发计划"中医药现代化"重点专项(2023YFC3503225) （2023YFC3503225）
