谷氨酰胺不同作用时间对单个核细胞热休克蛋白70表达的影响

Effect of Glutamine at Different Stimulation Times on Expression of Heat Shock Protein 70 in Peripheral Blood Mononuclear Cell

目的探讨经内毒素(lipopolysaccharide,LPS)预处理后,谷氨酰胺(glutamine,Gln)不同作用时间对单个核细胞(peripheral blood mononuclear cell,PBMC)热休克蛋白70(HSP70)表达的影响。方法取健康志愿者新鲜外周血滤过的血盘,用0.9%氯化钠注射液按1∶1稀释血液,Ficoll-泛影酸钠(Ficoll-Hypaque)密度梯度离心法提取PBMC,以LPS刺激PBMC2 h后,使用10 mmol/L的Gln分别刺激PBMC 0、0.5、1、2、3 h后提取蛋白,采用Western blot方法检测PBMC中HSP70表达情况。结果 PBMC经LPS刺激2 h后,予Gln分别作用0、0.5、1、2、3 h,结果显示作用1 h后,HSP70的表达量最高,与其他时间段比较差异均有统计学意义(P<0.05),随后表达量减少;作用2、3 h后表达量虽有减少,但与作用0.5 h比较,差异亦有统计学意义(P<0.05)。结论早期补充Gln可上调LPS诱导的PBMC中HSP70的表达,Gln发挥作用快速,但作用时间短。

Objective To explore the effect of glutamine （Gln） at different stimulation times on expression of heat shock protein 70 （HSP70） in peripheral blood mononuclear cell （PBMC） by lipopolysaccharide （LPS） pretreatment.Methods Fresh peripheral blood filtered through the blood discs of healthy volunteers were collected,then the blood on discs were diluted by 0.9％ sodium chloride injection （1∶ 1）,and PBMC was extracted by using density gradient centrifugation method with Ficoll Hypaque.PBMC was stimulated with 10 mmol/L Gln for 0,0.5,1,2 and 3 h after it was stimulated for 2 h with LPS.Expressions of HSP70 in PBMC were detected by Western blot after albumen extraction.Results PBMC was stimulated with Gln for 0,0.5,1,2 and 3 h after it was stimulated for 2 h with LPS.The highest expression of HSP70 in Gln stimulation for 1 h,and the differences were statistically significant compared with those in other time periods （P ＜ 0.05） ; then expression began to decrease.The differences were also statistically significant between stimulated for 2 and 3 h with stimulated for 0.5 h （P ＜ 0.05）.Conclusion Gln supplement in early period can upregulate the expression of HSP70 in PBMC induced by LPS,and Gln may respond rapidly with a short action time.