姜黄素对过氧化氢诱导的罗非鱼肝细胞热休克蛋白基因表达的影响

Effect of Curcumin on HSP Expression in Tilapia Hepatocytes under Hydrogen Peroxide Stress

【目的】探究热休克蛋白基因在氧化应激和姜黄素作用下的表达差异,了解姜黄素药效的发挥与热休克蛋白基因表达变化的关系,进一步探讨热休克蛋白对氧化应激条件下鱼类肝细胞中的功能,为鱼类氧化应激的毒理学机制及姜黄素的药理学机制提供理论资料。【方法】对24 h姜黄素预处理(保护组)与无姜黄素预处理(对照组)的罗非鱼肝细胞分别进行0、1、2.5 h的H_(2)O_(2)应激,并对肝细胞5种高分子热休克蛋白基因(HSP70、HSP90α、HSP90β、HSP60、HSP75)和5种小分子热休克蛋白基因(HSP30、HSPB1、HSPB7、HSPB8、HSPB11)的表达水平进行qPCR检测。【结果】与不经H_(2)O_(2)处理(0 h)相比,H_(2)O_(2)作用1 h能显著提升HSPB1表达量,降低HSP30表达量;H_(2)O_(2)作用2.5 h能显著提升HSP90a、HSPB1的表达量。姜黄素预处理24 h能显著提升HSP70、HSP90α、HSP30、HSPB1的表达量;姜黄素预处理24 h后再进行H_(2)O_(2)应激1 h时,HSP70、HSP90α、HSP30的表达量显著高于H_(2)O_(2)单独应激(对照组);姜黄素预处理24 h后再进行H_(2)O_(2)应激2.5 h时,HSP90α、HSP30、HSPB1的表达量显著高于H_(2)O_(2)单独应激。其余热休克蛋白在试验过程中无显著差异。【结论】H_(2)O_(2)单独应激可引起罗非鱼肝细胞HSP90a、HSP30、HSPB1基因的表达量变化,而姜黄素预处理能提高多种热休克蛋白基因的表达,提高了罗非鱼肝细胞的抗氧化能力,最终抵御H_(2)O_(2)引起的氧化应激并维持细胞活力。

【Objective】Expression of heat shock protein(HSP)gene under oxidative stress in the presence of curcumin was studied to understand the toxicological function of the protein and the pharmacological mechanism of curcumin in hepatocytes of Oreochromis nilotica.【Methods】Tilapia hepatocytes and those pretreated with curcumin for 24 h were subjected to H_(2)O_(2) stress for 0,1,or 2.5 h.The expressions of HSP70,HSP90α,HSP90β,HSP60,and HSP75,as well as 5 small molecule HSP genes,i.e.,HSP30,HSPB1,HSPB7,HSPB8,and HSPB11,in the cells were determined by qPCR.【Results】The H_(2)O_(2) treatment for 1 h significantly increased the expression of HSPB1 and decreased that of HSP30 in the tilapia hepatocytes,while the treatment that lasted for 2.5 h significantly increased the expressions of HSP90a and HSPB1.A pretreatment of curcumin for 24 h on the cells not only significantly elevated the expressions of HSP70,HSP90α,HSP30,and HSPB1 over control but also on those of HSP70,HSP90α,and HSP30 under 1 h H_(2)O_(2) stress,as well as those on HSP90α,HSP30,and HSPB1 under 2.5 h H_(2)O_(2) stress.However,no significant differences in other HSPs were observed.【Conclusion】As a spontaneous response of tilapia hepatocytes to oxidative stress,H_(2)O_(2) affected significantly the expressions of HSP90a,HSP30,and HSPB1 in the cells.By pretreating the hepatocytes with curcumin,expressions of these genes could be uplifted boosting the cellular antioxidant capacity to better resist the stress and maintain viability.