大鼠空白血清对阴虚内热型SLE患者外周血单核细胞蛋白质表达谱的影响

目的:在蛋白表达谱水平探讨大鼠空白血清作为含药血清空白对照的价值.方法:收集SLE阴虚内热证患者外周血单核细胞,分为实验组和对照组,实验组按孵化液量的20%加入大鼠空白血清,对照组按孵化液量的20%加入RPMI 1640培养液,收集孵化后细胞进行PMBC的蛋白表达谱检测.结果:实验组和对照组比较,外周血单核细胞的32个蛋白峰表达有差异,其中11个蛋白峰表达有统计学差异;建立了质荷比(m/z)为7027、5270、2908、4128、2311Da模式来判别大鼠空白血清和对照之间的差别.应用生物信息学方法分析7027Da模式蛋白,可知其对应的蛋白为HLADRB1,相应的基因为HLADRB1.结论:大鼠空白血清本身对SLE阴虚内热证患者外周血单核细胞具有明确的调节作用,因此分析含药血清作用时应考虑到此作用.

Investigation on Effects of Normal Rat Serum as Controls on Testing Protein Expression Profile of PBMC from Patients with Internal Heat due to Yin Deficiency Type SLE

Objective：To investigate the practicality of normal rat serum to be used as a nega- tive control of medicine-loaded serum in testing protein expression profile, nethods：PBMC were collected from patients with internal heat due to Yin deficiency type SLE. PBMC of experimental group were cultured with 20% rat serum, while PBMC of control group were cultured with 20% RPMI 1640 medium. SELDI-TOF-MS was adopted to detect protein expression profile of PBMC af- ter incubation. Results： PBMC of experimental group showed differential expression of thirty-two proteins compared with control group, 11 of which showed statistically significant difference. The potential protein biomarkers of 7027,5270,2908,4128 tection between two groups. By bioinformaties method ,2311Da were established for difference detection between two groups. By bioinformatics method the protein with peak mass/charge ranos of 7027Da was identified as HLADRB1 protein coded by HLADRB1 gene. Conclusion： Protein expression profiling studies have shown that normal rat serum itself had regulation function on protein expression profile by PBMC of SLE patients. It is necessary to take it into consideration when we analyze the effect of medicine-loaded serum.