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目的研究热休克(heat shock,HS)对人单核细胞株THP-1细胞IL-37表达及其对促炎细胞因子产生的影响。方法对THP-1细胞进行42℃热休克处理1 h,然后分别于37℃恢复培养0~6 h,用流式细胞术检测胞内IL-37的表达。对THP-1细胞用热休克处理与脂多糖(lipopolysaccharides,LPS)刺激,分别收集细胞及其培养上清,用流式细胞术检测各组细胞IL-37表达,ELISA检测各组细胞培养上清中促炎细胞因子的水平。结果流式细胞术检测结果显示,经42℃热休克处理后THP-1细胞于37℃分别恢复培养0、2、4、6 h,细胞IL-37的阳性表达率分别比对照组增高21.1、30.6、34.1、15.1倍,每个细胞表达IL-37的平均荧光强度(Mean Fluorescence Intensity,MFI)分别比对照组增高4.6、7.5、8.3、3.7倍。此外,还发现THP-1细胞单独用热休克处理或LPS刺激后,其IL-37表达水平无显著差异,但热休克处理结合LPS刺激的细胞IL-37表达水平则分别显著高于单独用热休克处理或LPS刺激的细胞,其MFI分别比单独用热休克处理或LPS刺激细胞增高2.0、1.9倍。ELISA结果显示,LPS刺激后THP-1细胞产生炎症因子IL-1β、TNF-α、IL-6的水平分别比对照组高2.9、2.1、3.3倍,而热休克处理结合LPS刺激的细胞的IL-1β、TNF-α与IL-6的水平则比仅用LPS刺激的细胞分别降低了75.1%、69.6%、66.4%。结论热休克能上调THP-1细胞IL-37表达,其上调表达可能参与抑制促炎细胞因子的产生。
Objective To investigate the effect of heat shock (HS) on the expression of IL-37 in human monocytic THP-1 cells and its effect on proinflammatory cytokines. THP-1 cells were heat-shocked at 42 ℃ for 1 h, then resumed culture at 37 ℃ for 0-6 h. The expression of IL-37 in cells was detected by flow cytometry. THP-1 cells were stimulated with heat shock and lipopolysaccharides (LPS), and the cells and their culture supernatants were collected. The expression of IL-37 in each group was detected by flow cytometry. The expression of IL-37 In the level of pro-inflammatory cytokines. Results The results of flow cytometry showed that the THP-1 cells were resumed culture at 37 ℃ for 4 days at 37 ℃, respectively. The positive rate of IL-37 in THP-1 cells increased by 21.1% 30.6, 34.1 and 15.1 times respectively. Mean Fluorescence Intensity (MFI) of IL-37 in each cell was increased by 4.6, 7.5, 8.3 and 3.7 times respectively. In addition, THP-1 cells were also found no significant difference in IL-37 expression levels after heat shock alone or LPS stimulation, but heat shock treatment and LPS-stimulated cells IL-37 expression levels were significantly higher than the heat alone Shock-treated or LPS-stimulated cells showed an MFI increase of 2.0 and 1.9 times, respectively, compared to the cells treated with heat shock or LPS alone. The results of ELISA showed that the levels of IL-1β, TNF-α and IL-6 in THP-1 cells stimulated by LPS were 2.9, 2.1 and 3.3 times higher than that of the control group, respectively, while heat shock treatment combined with LPS-stimulated cells IL -1β, TNF-αand IL-6 were decreased by 75.1%, 69.6% and 66.4% respectively compared with those stimulated by LPS only. Conclusion Heat shock can up-regulate the expression of IL-37 in THP-1 cells, which may be involved in the inhibition of the production of proinflammatory cytokines.