1,25(OH)_(2)D_(3)联合NF-κB p65 siRNA保护肺炎链球菌感染的肺泡上皮细胞损伤

1,25(OH)_(2)D_(3)combined with NF-κB p65 siRNA protects alveolar epithelial cells from streptococcus pneumoniae infection

目的:探讨1,25(OH)_(2)D_(3)与核转录因子-κB(NF-κB)p65 siRNA单独或联合作用对肺炎链球菌(SP)感染的肺泡上皮细胞增殖、凋亡的影响。方法:分别使用1,25(OH)_(2)D_(3)、NF-κB p65 siRNA及二者联合处理SP感染的肺泡上皮细胞A549,实时荧光定量PCR(qRT-PCR)检测NF-κB p65 mRNA表达,Western blot检测NF-κB p65、Bcl2相关X蛋白(Bax)、B细胞淋巴瘤/白血病2(Bcl2)、IL-1β、IL-6、TNF-α、NF-κB p-p65蛋白表达,MTT比色法和流式细胞术分别检测细胞活力与凋亡。结果:SP明显促进A549细胞中NF-κB p65 mRNA、NF-κB p65蛋白表达、细胞凋亡率、Bax、IL-1β、IL-6、TNF-α、NF-κB p-p65表达(P<0.05),显著抑制相对细胞活力、Bcl2表达(P<0.05)。1,25(OH)_(2)D_(3)、NF-κB p65 siRNA以及1,25(OH)_(2)D_(3)联合NF-κB p65 siRNA处理均明显抑制SP感染的A549细胞中NF-κB p65 mRNA、NF-κB p65蛋白表达、细胞凋亡、Bax、IL-1β、IL-6、TNF-α、NF-κB p-p65表达(P<0.05),提高细胞活力、Bcl2表达(P<0.05),并且1,25(OH)_(2)D_(3)联合NF-κB p65 siRNA对SP损伤A549细胞增殖、抑制细胞凋亡的作用更强。结论:1,25(OH)_(2)D_(3)联合NF-κB p65 siRNA对SP感染的肺泡上皮细胞损伤具有保护作用,且效果优于1,25(OH)_(2)D_(3)或NF-κB p65 siRNA单独作用,机制可能与调控炎症因子、NF-κB信号通路有关。

Objective:To investigate the effects of 1,25(OH)_(2)D_(3)and nuclear factor-κB(NF-κB)p65 siRNA alone or in combination on the proliferation and apoptosis of alveolar epithelial cells infected with streptococcus pneumoniae(SP).Methods:SP-infected alveolar epithelial cells A549 were treated with 1,25(OH)_(2)D_(3),NF-κB p65 siRNA and both,NF-κB p65 mRNA expression was detected by qRT-PCR.Western blot analyzed expressions of NF-κB p65,Bax,Bcl2,IL-1β,IL-6,TNF-α,NF-κB p-p65 protein,cell viability and apoptosis were detected by MTT colorimetry and flow cytometry.Results:SP significantly promoted the expression of NF-κB p65 mRNA,NF-κB p65 protein,apoptosis rate,Bax,IL-1β,IL-6,TNF-αand NF-κB p-p65 in A549 cells(P<0.05),obviously inhibited relative cell viability,Bcl2 expression(P<0.05).1,25(OH)_(2)D_(3),NF-κB p65 siRNA and 1,25(OH)_(2)D_(3)combined with NF-κB p65 siRNA treatment remarkably inhibited the expression of NF-κB p65 mRNA,NF-κB p65 protein,cell apoptosis,Bax,IL-1β,IL-6,TNF-α,NF-κB p-p65 expression in SP-infected A549 cells(P<0.05),increased cell viability,Bcl2 expression(P<0.05),and 1,25(OH)_(2)D_(3)combined with NF-κB p65 siRNA showed a stronger effect on promotiing cell proliferation and inhibiting cell apoptosis in SP-infected A549 cells.Conclusion:1,25(OH)_(2)D_(3)combined with NF-κB p65 siRNA protects on alveolar epithelial cell injury induced by SP infection,and the effect is better than that of 1,25(OH)_(2)D_(3)or NF-κB p65 siRNA alone.The mechanism might be related to the regulation of inflammatory factors and NF-κB signaling pathways.