胆囊收缩素通过胆囊收缩素受体A减弱炎症反应降低脂多糖诱导的急性肺损伤  

Cholecystokinin Attenuates Inflammatory Reaction and Alleviates Lipopolysaccharide-induced Acute Lung Injury Through CCKAR

作　　者：谭媛[1] 廖勇杰 岳嗣凤 Tan Yuan;Liao Yongjie;Yue Sifeng(Department of Neonates,Affiliated Hospital of Guilin Medical University,Guilin 541001,China)

机构地区：[1]桂林医学院附属医院新生儿科,桂林541001

出　　处：《华中科技大学学报（医学版）》2022年第5期625-630,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：广西壮族自治区卫生健康委员会科研课题(No.Z20190648)。

摘　　要：目的探讨胆囊收缩素(CCK)和胆囊收缩素受体A(CCKAR)在急性肺损伤(ALI)中的作用及其潜在分子机制。方法使用GEO2R工具分析来自GEO数据库的2个数据集(GSE18341,GSE2411)表达数据,以筛选小鼠正常组织和ALI组织中的差异表达基因(DEGs)。利用脂多糖(LPS)诱导BEAS-2B细胞构建体外ALI模型,将其分为空白对照组(Control)、LPS处理组(LPS)、CCKAR过表达阴性对照组(LPS+OE-NC)、CCKAR过表达组(LPS+OE-CCKAR)、CCK过表达组(LPS+OE-CCK)和CCK与CCK拮抗剂组(LPS+CCK+Proglumide)。采用RT-PCR及Western blot检测基因表达情况;MTT法检测细胞活力;ELISA检测细胞上清IL-1β、IL-6和TNF-α的水平。结果筛选获得80个共同DEGs,其中CCKAR下调最为显著。相较于Control组,LPS组CCKAR表达下调(P<0.01),细胞活力显著降低(P<0.01),炎症因子IL-1β、IL-6和TNF-α水平升高(均P<0.01)。CCK提高了LPS诱导的BEAS-2B细胞中CCKAR的表达(P<0.01),CCKAR或CCK过表达逆转了LPS对细胞活力及炎症因子水平的影响(均P<0.05),Proglumide逆转了CCK对细胞活力和炎症因子水平的影响。相较于Control组,LPS组p-p65/p65以及p-IκB/IκB表达上调(均P<0.01),CCKAR或CCK过表达后,p-p65/p65以及p-IκB/IκB表达下调(均P<0.01),Proglumide逆转了CCK对p-p65/p65以及p-IκB/IκB表达的作用。结论CCK通过CCKAR抑制LPS诱导的细胞损伤和炎症反应,并可能通过调节NF-κB信号通路发挥作用。Objective To investigate the role and underlying molecular mechanisms of cholecystokinin(CCK)and CCK receptor A(CCKAR)in acute lung injury(ALI).Methods The GEO2R tool was used to analyze the expression data of two datasets(GSE18341,GSE2411)from GEO database to screen out differentially expressed genes(DEGs)in normal and ALI tissues from mice.Lipopolysaccharide(LPS)was employed to treat BEAS-2B cells for the construction of in vitro ALI model.Cells were classified into normal group(Control),model group(LPS),CCKAR overexpression negative control group(LPS+OE-NC),CCKAR overexpression group(LPS+OE-CCKAR),CCK overexpression group(LPS+OE-CCK),and antagonist group(LPS+CCK+Proglumide).RT-PCR and Western blot were used to detect the expression of genes.MTT assay was applied to assess cell viability.ELISA was performed to determine the levels of IL-1β,IL-6 and TNF-αin the cell supernatants.Results Eighty common DEGs were obtained by screening,among which CCKAR was the most significantly downregulated gene.Compared to Control group,CCKAR expression declined(P<0.01),cell viability was markedly decreased(P<0.01)and inflammatory factors IL-1β,IL-6 and TNF-αlevels were elevated in LPS group(all P<0.01).Moreover,CCK treatment elevated the expression of CCKAR in LPS-induced BEAS-2B cells(P<0.01).However,CCKAR or CCK overexpression reversed the effect of LPS on cell viability and inflammation levels(both P<0.05).Further,the effects of CCK on cell viability and inflammatory factors were counteracted by Proglumide.Additionally,expression levels of p-p65/p65 and p-IκB/IκB were upregulated in LPS group compared with the control group(both P<0.01).However,expression levels of p-p65/p65 and p-IκB/IκB were downregulated after CCKAR or CCK overexpression(both P<0.01),and Proglumide reversed the impacts of CCK on p-p65/p65 and p-IκB/IκB expression.Conclusion CCK inhibits LPS-induced cell injury and inflammation in BEAS-2B cells through CCKAR,which may function by regulating the NF-κB signaling pathway.

关 键 词：胆囊收缩素 脂多糖 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]