【摘要】 目的:探讨热休克因子1(heat shock factor 1,HSF1)对热应激所致Raw2 6 4.7巨噬细胞凋亡的影响。方法:采用热应激(4 2.5℃±0.5℃)处理稳定表达小鼠HSF1基因的Raw2 6 4.7巨噬细胞1h,3 7℃分别恢复6,9,1 2,2 4 h,采用流式细胞术,hoechst3 3 2 5 8染色和DNA琼脂糖凝胶电泳检测细胞凋亡。结果:流式细胞术结果显示,热应激后对照组(转空载体)细胞凋亡核百分率较热应激前明显升高,9 h达峰值(约为6 0%),此时荧光染色可见3 0%的细胞出现核固缩,凋亡小体等典型的凋亡形态学改变;并于热应激后6,9,1 2 h均能检测到清晰的DNA梯状条带。与转空载体对照组相比,HSF1过表达能显著降低热应激所致凋亡及明显抑制DNA的断裂。结论:HSF1可以抑制热应激所致的Raw2 6 4.7巨噬细胞凋亡。更多还原

【Abstract】 Objective To observe the effect of heat shock factor 1(HSF1) on heat stress-induced apoptosis in Raw264.7 macrophages.Methods Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1HSF1 were exposed to heat stress(42.5℃±0.5℃) for 1h and recovered at 37℃ for 6,9,12,and 24h respectively.Flow cytometry(FCM),Hoechst 33258 staining and DNA ladder assays were performed to assess the apoptosis.Results After heat stress,FCM showed that apoptotic cells were increased significantly and reached the peak at 9 h in Raw 264.7 cells transfected with pcDNA3.1,and were characterized with classical morphologic changes including apoptotic body and nuclear condensation.Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly at 6,9,and 12 h after the heat stress.But the overexpression of HSF1 could reduce the number of apoptotic cells and inhibit DNA fragmentation.Conclusion HSF1 can inhibit heat stress-induced apoptosis in Raw264.7 macrophages.更多还原