摘要
目的分析颗粒酶B在PM_(2.5)促进自然杀伤(natural killer,NK)细胞炎症因子表达中的作用。方法(1)通过设置PM_(2.5)染毒剂量梯度,分组为空白对照组(0μg/ml)、PM_(2.5)(50μg/ml)组、PM_(2.5)(150μg/ml)组和PM_(2.5)(450μg/ml)组,观察PM_(2.5)暴露与NK细胞颗粒酶B和炎症因子TNF-α、IL-1β表达间有无相关性;(2)为明确PM_(2.5)上调NK细胞炎症因子表达中颗粒酶B是否发挥促进作用,根据是否行PM_(2.5)染毒和颗粒酶B抑制剂Serpina3n预处理,实验分为空白对照组、Serpina3n组、PM_(2.5)组和Serpina3n+PM_(2.5)组。在PM_(2.5)染毒前1h,Serpina3n组和Serpina3n+PM_(2.5)组加入Serpina3n(100 ng/ml);(3)为明确在PM_(2.5)上调NK细胞炎症因子表达中颗粒酶B是否通过IL-18发挥作用,根据是否行PM_(2.5)染毒和IL-18抑制剂IL-18BP预处理,实验分为空白对照组、IL-18BP、PM_(2.5)组、IL-18BP+PM_(2.5)组。在PM_(2.5)染毒前1 h,IL-18BP组和IL-18BP+PM_(2.5)组加入IL-18BP(100 ng/ml)。分别检测每组NK细胞蛋白表达情况和细胞培养液TNF-α、IL-1β水平。结果PM_(2.5)促进了NK细胞颗粒酶B和炎症因子TNF-α、IL-1β表达;阻断颗粒酶B抑制NK细胞IL-18表达和NF-κB磷酸化,降低TNF-α和IL-1β表达;阻断IL-18抑制了NF-κB磷酸化,降低TNF-α和IL-1β表达。结论PM_(2.5)可能通过激活颗粒酶B/IL-18信号通路促进NK细胞TNF-α和IL-1β表达,机制可能与激活NF-κB信号通路有关。
Objective To explore the role of granzyme B in PM_(2.5)promoting the expression of inflammatory cytokines in natural killer(NK)cells.Methods First,the PM_(2.5)dose gradient was set.The experiment was divided into blank control group(0μg/ml),PM_(2.5)group(50μg/ml),PM_(2.5)group(150μg/ml),and PM_(2.5)group(450μg/ml).By setting the PM_(2.5)dose gradient,we observed the correlation between the expression of NK cell granzyme B and inflammatory cytokines TNF-αand IL-1β.Second,in order to determine whether granzyme B plays a promoting role in the expression of inflammatory cytokines in NK cells promoted by PM_(2.5),the experiment was divided into blank control group,Serpina3n group,PM_(2.5)group and Serpina3n+PM_(2.5)group according to whether PM_(2.5)exposure and Serpina3n inhibitor were pretreated.Serpina3n group and Serpina3n+PM_(2.5)group were added Serpina3n(100 ng/ml)1 hour before PM_(2.5)poisoning.Third,in order to determine whether granzyme B plays a role through IL-18 in PM_(2.5)promoting the expression of inflammatory cytokines in NK cells,the experiment was divided into blank control group,IL-18BP,PM_(2.5)group,and IL-18BP+PM_(2.5)group according to whether PM_(2.5)exposure and IL-18BP inhibitor were pretreated.One hour before PM_(2.5)exposure,IL-18BP group and IL-18BP+PM_(2.5)group were added with IL-18BP(100 ng/ml).The expression of NK cell protein and the levels of TNF-αand IL-1βin cell culture medium were detected in each group separately.Results PM_(2.5)promoted the expression of granzyme B and inflammatory factors TNF-αand IL-1βin NK cells.Blocking granzyme B inhibited IL-18 expression and NF-κB phosphorylation,and decreased TNF-αand IL-1βexpression in NK cells.Blocking IL-18 inhibited the phosphorylation of NF-κB and decreased the expression of TNF-αand IL-1β.Conclusion PM_(2.5)may promote the expression of TNF-αand IL-1βin NK cells by activating granzyme B/IL-18 signaling pathway,and the mechanism may be related to the activation of NF-κB signaling pathway.
作者
陈向军
王在强
王博荣
王莉
方芳
金发光
王光辉
Chen Xiangjun;Wang Zaiqiang;Wang Borong;Wang Li;Fang Fang;Jin Faguang;Wang Guanghui(The second Department of Intensive Care Medicine,Xi′an Chest Hospital,Xi′an 710100,China;Department of Respiratory and Critical Care Medicine,The Second Affiliated Hospital of Air Force Medical University,Xi′an 710038,China)
出处
《中华肺部疾病杂志（电子版）》
2024年第2期207-211,共5页
Chinese Journal of Lung Diseases(Electronic Edition)
基金
陕西省重点研发计划(2018ZDCXL-SF-02-03-02)。
