阿魏酸抑制NLRP3炎症小体减轻辐射诱导的AHH-1细胞炎症

目的:研究阿魏酸(Ferulic Acid)对辐射所致AHH-1细胞炎症反应的作用及机制。方法:以0 Gy^8 Gy^60Coγ射线照射AHH-1细胞,照后24 h检测细胞活力,确定适宜的照射剂量。采用酶联免疫法(ELISA)和蛋白免疫印迹法(Western Blot)检测γ射线照射后不同时间点AHH-1细胞中炎症因子白介素1β(IL-1β)和白介素18(IL-18)的表达。CCK-8法检测不同浓度阿魏酸对AHH-1细胞活力的影响,确定适宜的阿魏酸给药浓度。阿魏酸预处理,以4 Gyγ射线照射AHH-1细胞建立炎症反应模型,并于照后12 h提取细胞总蛋白,采用Western Blot法检测IL-1β、IL-18、NLRP3和Caspase-1蛋白表达水平的变化。结果:与0 Gy组比较,AHH-1细胞受到4 Gyγ射线照射后24 h,细胞活力约下降至60%。4 Gyγ射线照射后,随着时间的增加,炎症因子含量和炎症因子表达增多,照后12 h蛋白表达最显著(P <0. 05或P <0. 01)。与正常对照组相比,0. 01μmol/L^200μmol/L的阿魏酸单独给药,对AHH-1细胞活力无明显影响;与模型对照组比较,阿魏酸0. 01、0. 1、1μmol/L预处理组细胞活力明显升高(P <0. 05或P <0. 01),1μmol/L阿魏酸可显著抑制炎症因子IL-1β和IL-18的分泌量,且使细胞中IL-1β、IL-18、NLRP3和Caspase-1蛋白表达水平显著降低。结论:阿魏酸对辐射诱导的AHH-1细胞炎性反应有保护作用,可能与抑制NLRP3炎症小体有关。

Ferulic Acid Alleviates Radiation-Induced AHH-1 Inflammatory Response by Inhibiting the NLRP3 Inflammasome

Objective: To investigate the effect and the mechanism of ferulic acid on AHH-1 cell inflammation induced by radiation. Methods:AHH-1 cells were irradiated with 0 ~ 8 Gy^60 Co gamma-ray. 24 hours after irradiation,the cell viability was evaluated to determine the appropriate radiation dose. The release of the inflammatory factor IL-1β and IL-18 in the supernatant of AHH-1 cell was detected by ELISA Kit and Western Blot at different time points after 4 Gy gamma-ray irradiation. The effects of different concentrations of ferulic acid on the viability of AHH-1 cells were detected by cell counting kit-8( CCK-8) assay to determine the appropriate concentration of ferulic acid. After the pretreatment with appropriate concentration of ferulic acid,AHH-1 cells were irradiated with 4 Gy gamma-ray to establish an inflammatory response model. The total protein was extracted 12 h after irradiation,and the expressions of IL-1β,IL-18,NLRP3 and caspase-1 proteins were evaluated by Western Blot. Results: Compared with the 0 Gy group,the cell activity of AHH-1 decreased to 60% about 24 h after 4 Gy gamma-ray irradiation. After 4 Gy dose irradiation,the secretion of inflammatory factors increased as time prolonged,at 12 h after irradiation,the expression of protein showed the most significance. Compared with the control group,0. 01 ~ 200 mol/L ferulic acid had no significant effect on the activity of AHH-1 cells. Compared with the radiation group,the cell viability in the pretreatment with ferulic acid group was significantlyincreased,and the expression levels of IL-1β,IL-18,NLRP3 and caspase-1 proteins were significantly decreased. Conclusion: Ferulic acid has a protective effect on the radiation-induced inflammatory response in AHH-1 cells,which may be related to the inhibition of NLRP3 inflammasome.