慢性阻塞性肺疾病急性加重期患者血清对PM2.5所致MH-S细胞炎症的影响及salubrinal的作用

Effects of chronic obstructive pulmonary disease patients'serum and sa⁃lubrinal on inflammation of MH-S cells induced by PM2.5

目的:探讨慢性阻塞性肺疾病急性加重期(AECOPD)患者血清和salubrinal在细颗粒物(PM2.5)诱导的小鼠肺泡巨噬细胞株MH-S炎症中的作用。方法:(1)将MH-S细胞分为对照组、PM2.5组、AECOPD患者血清组、健康志愿者血清组、PM2.5+AECOPD患者血清组和PM2.5+健康志愿者血清组,检测组蛋白脱乙酰酶2(HDAC2)活性,细胞上清液中白细胞介素17(IL-17)浓度,分析IL-17与HDAC2活性间的相关性;(2)进一步探究salubrinal对PM2.5刺激的MH-S细胞的作用,分为对照组、PM2.5组和salubrinal+PM2.5组,检测MH-S细胞中C/EBP同源蛋白(CHOP)和葡萄糖调节蛋白78(GRP78)表达、HDAC2活性及细胞上清液中IL-17浓度。结果:(1)PM2.5组、AECOPD患者血清组、健康志愿者血清组和PM2.5+AECOPD患者血清组和PM2.5+健康志愿者血清组较对照组MH-S细胞HDAC2活性均降低,IL-17分泌增加,且AECOPD患者血清组较健康志愿者血清组IL-17升高,MH-S细胞HDAC2活性降低更明显(P<0.05)。PM2.5+AECOPD患者血清组较PM2.5+健康志愿者血清组IL-17升高、HDAC2活性降低明显(P<0.05)。IL-17与HDAC2之间存在负相关(r=-0.786,P<0.01);(2)PM2.5刺激12 h后细胞较对照组CHOP荧光增强、核内居多,GRP78荧光增强、胞质明显,IL-17含量增加,HDAC2活性降低(P<0.05)。加入salubrinal干预后CHOP荧光较PM2.5组减弱,GRP78荧光较PM2.5组增强,IL-17含量降低,HDAC2活性增加(P<0.05)。结论:PM2.5刺激会导致MH-S细胞IL-17分泌增多,HDAC2活性降低,血清会加重PM2.5对MH-S细胞的影响;IL-17与HDAC2之间存在负相关;PM2.5刺激使CHOP和GRP78表达增多,激活MH-S细胞内质网应激;salubrinal可能通过下调MH-S细胞CHOP表达抑制了PM2.5对MH-S细胞炎症功能。

AIM:To investigate the role of acute exacerbation of chronic obstructive pulmonary disease(AECOPD)patients’ serum and salubrinal in PM2. 5-induced inflammatory function of mouse alveolar macrophages(MHS cells).METHODS:Six groups were set up:control group,PM2. 5 group,AECOPD serum group,healthy volunteer serum group,PM2. 5+AECOPD serum group,and PM2. 5+healthy volunteer serum group. The activity of histone deacetylase 2(HDAC2)and the concentration of interleukin-17(IL-17)in the supernatant of MH-S cells were measured. The correlation between IL-17 and HDAC2 activity was analyzed by Pearson correlation analysis. To further explore the effect of salubrinal on MH-S cell stimulated by PM2. 5,the cells were divided into control group,PM2. 5 group and salubrinal+PM2. 5 group,and stimulated for 12 h. The expression of C/EBP homologus protein(CHOP)and glucose-regulated protein 78(GRP78),the activity of HDAC2 and the concentration of IL-17 in the supernatant were measured.RESULTS:Compared with control group,the activity of HDAC2 was decreased and the secretion of IL-17 was increased in experimental group. The IL-17 concentration were higher and the HDAC2 activity were lower in PM2. 5 and AECOPD serum group compared with PM2. 5 and healthy volunteer serum group(P<0. 05). There was a negative correlation between IL-17 and HDAC2(r=-0. 786,P<0. 01). After stimulation with PM2. 5 for 12 h,the CHOP fluorescence was increased and enter into nucleus obviously,the GRP78 fluorescence was increased and significantly in cytoplasm,the IL-17 content was increased and the HDAC2 activity was decreased compared with control group(P<0. 05). After the intervention of salubrinal,the fluorescence of CHOP was lower than that of PM2. 5,the fluorescence of GRP78 was higher than that of PM2. 5,the content of IL-17 was decreased,and the activity of HDAC2 was increased. The differences were statistically significant(P<0. 05).CONCLUSION:PM2. 5 increases the secretion of IL-17 and decreases the activity of HDAC2 in MH-S cells. Serum aggravates the effect of PM2. 5 on MH-S cells. There is a negative correlation between IL-17 and HDAC2.PM2. 5 stimulation increases the expression of CHOP and GRP78,and activates the endoplasmic reticulum stress. Salubrinal may inhibit the inflammatory effect of PM2. 5 on MH-S cells by inhibiting the expression of CHOP.