硼替佐米调节NF-κB途径对IL-1β诱导的ATDC5细胞炎症性损伤的影响研究

Effect of bortezomib on the NF-κB pathway and IL-1βinduced inflammatory damage in ATDC5 cells

目的探究硼替佐米(Bor)对白介素-1β(IL-1β)诱导的ATDC5细胞炎症性损伤及核因子κB(NF-κB)的影响。方法0、5、15、25、35、45、55 nmol/L Bor处理ATDC5细胞48 h,CCK-8实验筛选无细胞毒性浓度,0、1.0、5.0、12.5、25.0 nmol/L Bor分别与10μg/mL IL-1β共培养ATDC5细胞48 h,CCK-8法检测细胞增殖能力;流式细胞术检测细胞凋亡情况;酶联免疫吸附(ELISA)检测上清液中IL-1β表达情况;蛋白免疫印迹检测细胞中NF-κB、B细胞淋巴瘤/白血病-2原癌基因(BcL-2)、BcL-2相关X蛋白(BAX)蛋白水平。结果0、5、15、25 nmol/L Bor组差异无统计学意义(P>0.05)。与0 nmol/L Bor组相比,35、45、55 nmol/L Bor组细胞存活率降低(P<0.05)。Bor浓度≤25 nmol/L对该细胞无毒。与0 nmol/L Bor组相比,IL-1β组细胞存活率、细胞中BcL-2蛋白水平降低(P<0.05),凋亡率、上清液中IL-1β水平、细胞中NF-κB、BAX蛋白水平升高(P<0.05)。随着Bor剂量的增加,细胞存活率、细胞中BcL-2蛋白水平逐渐升高(P<0.05),细胞凋亡率、上清液中IL-1β、细胞中NF-κB、BAX蛋白水平逐渐降低(P<0.05),呈剂量依赖效应。结论Bor可抑制细胞凋亡及炎症因子水平从而减弱IL-1β诱导的ATDC5细胞炎症性损伤,可能是通过抑制NF-κB途径实现的。

Objective To investigate the effects of bortezomib(Bor)on nuclear factorκB(NF-κB)and inflammatory damage in ATDC5 cells induced by interleukin-1β(IL-1β).Methods ATDC5 cells were treated with(0,5,15,25,35,45,55)nmol/L Bor for 48 hours,and CCK-8 was used to determine the cytotoxic concentration of Bor.ATDC5 cells were co-cultured for 48 hours with(0,1.0,5.0,12.5,25.0)nmol/L Bor and 10μg/mL IL-1β.CCK-8 was used to detect cell proliferation and apoptosis was detected by flow cytometry.IL-1βin supernatant was detected using an enzyme-linked immunosorbent assay(ELISA),and the levels of NF-κB,B-cell lymphoma/leukemia-2 proto oncogene(BcL2)and BcL2-related X protein(BAX)were determined by Western blot.Results There was no significant difference in(0,5,15,25)nmol/L Bor group(P>0.05).Compared with the 0 nmol/L Bor group,the cell survival rate of the 35,45,and 55 nmol/L Bor groups was reduced(P<0.05).A concentration of≤25 nmol/L Bor was non-toxic to the cells.Compared with the 0 nmol/L Bor group,the cell survival rate and BcL2 protein level in the IL-1βgroup was decreased(P<0.05),and the apoptosis rate,IL-1βlevel in the supernatant,and NF-κB and BAX protein levels were increased(P<0.05).With increasing Bor dose,the cell survival rate and the level of BcL2 protein in cells gradually increased(P<0.05),while the apoptosis rate,the level of IL-1βin supernatant,and the levels of NF-κB and BAX proteins in cells gradually decreased(P<0.05)in a dose-dependent manner.Conclusions Bor can inhibit apoptosis and reduce the levels of inflammatory factors,thereby reducing inflammatory damage to ATDC5 cells induced by IL-1β,which may occur via inhibition of the NF-κB pathway.