LPS通过促进FGFR1磷酸化诱导肺上皮细胞炎症反应及凋亡  

LPS induces inflammatory response and apoptosis in pulmonary epithelial cells by promoting the phosphorylation of FGFR1

作　　者：吴跃明[1] 楼天正[1] 徐俊龙[1] WU Yueming;LOU Tianzheng;XU Junlong(Department of Critical Care Medicine,Lishui People's Hospital,Lishui 323000,Zhejiang,China)

机构地区：[1]丽水市人民医院重症医学科,浙江丽水323000

出　　处：《临床检验杂志》2023年第3期235-240,共6页Chinese Journal of Clinical Laboratory Science

基　　金：浙江省基础公益研究计划(LGF20H15008)。

摘　　要：目的 探讨脂多糖(LPS)通过促进成纤维细胞生长因子受体1(FGFR1)磷酸化的方式诱导肺上皮细胞炎症反应及凋亡。方法 培养肺上皮Beas-2B细胞并进行以下分组处理,以不含药物的培养基处理作为对照组,用5 mg/L LPS刺激作为LPS组,另加入不同浓度FGFR1选择性抑制剂AZD4547(2.5、5.0、10.0μmol/L)作为AZD4547组。采用CCK8法检测各组细胞的增殖活力,Tunel法检测细胞凋亡率,ELISA法检测细胞培养基中TNF-α、IL-1β、IL-6的含量,Western blot检测各组细胞总蛋白中磷酸化成纤维细胞生长因子受体1(p-FGFR1)、B淋巴细胞瘤-2基因(Bcl-2)蛋白、Bcl-2相关X蛋白(Bax)、切割型半胱氨酸天冬氨酸蛋白水解酶-3(cleaved caspase-3)以及核蛋白中p65核因子-κB(NF-κB)的表达水平。结果 LPS组Beas-2B细胞的增殖活力及Bcl-2水平均显著低于对照组(t分别为11.421、17.784,P<0.05),而Beas-2B细胞的细胞凋亡率、p-FGFR1、Bax、cleaved caspase-3、p65 NF-κB水平及培养基中TNF-α、IL-1β、IL-6含量均高于对照组(t分别为9.126、11.329、12.684、13.147、15.027、11.621、8.925、9.726,P<0.05);2.5、5.0、10.0μmol/L浓度AZD4547组Beas-2B细胞的增殖活力及Bcl-2水平均显著高于LPS组(F分别为45.012、72.286,P<0.05),而细胞凋亡率、p-FGFR1、Bax、cleaved caspase-3、 p65 NF-κB水平及培养基中TNF-α、IL-1β、IL-6含量均显著低于LPS组(F分别为43.527, 94.166, 113.028, 102.515, 41.091, 51.301, 30.280,P<0.05),且AZD4547浓度越高,上述变化趋势越显著。结论 LPS诱导肺上皮细胞炎症反应及凋亡激活的作用可能与促进FGFR1磷酸化有关。Objective To investigate whether lipopolysaccharide(LPS)may induce inflammatory response and apoptosis in pulmonary epithelial cells by promoting the phosphorylation of fibroblast growth factor receptor 1(FGFR1).Methods Lung epithelial Beas-2B cells were cultured and divided into the control group with drug-free medium,LPS group stimulated with 5 mg/L LPS,and AZD4547 groups treated with different concentrations of AZD4547(2.5,5.0,10.0μmol/L),a selective inhibitor of the FGFR1.The proliferation activity and apoptosis rate of Beas-2B cells in each group were determined by the CCK8 method and Tunel assay,respectively.The concentrations of TNF-α,IL-1βand IL-6 in culture medium were detected by ELISA.The expression levels of phosphorylated FGFR1(p-FGFR1),Bcl-2,Bax,cleaved caspase-3 and p65 NF-κB in Beas-2B cells of each group were determined by Western blot.Results The proliferation activity and Bcl-2 levels of Beas-2B cells in the LPS group were significantly lower than those in the control group(t=11.421 and 17.784,P<0.05),while the apoptosis rate,the expression levels of p-FGFR1,cleaved caspase-3,p65 NF-κB and Bax2,and the concentrations of TNF-α,IL-1βand IL-6 in culture medium were significantly higher than those in the control group(t=9.126,11.329,12.684,13.147,15.027,11.621,8.925 and 9.726,respectively,P<0.05).The proliferation activity and Bcl-2 levels of Beas-2B cells treated with different concentrations of AZD4547(2.5,5.0,10.0μmol/L)were significantly higher than those in the LPS group(F=45.012 and 72.286,P<0.05),while the apoptosis rate,the expression levels of p-FGFR1,Bax2,cleaved caspase-3 and p65 NF-κB,and the concentrations of TNF-α,IL-1βand IL-6 in culture medium were significantly lower than those in the LPS group(F=43.527,94.166,113.028,102.515,41.091,51.301 and 30.280,respectively,P<0.05).The higher the concentration of AZD4547,the more significant the above changes.Conclusion The effect of LPS inducing inflammatory response and apoptosis of pulmonary epithelial cells may be relate to the

关 键 词：成纤维细胞生长因子受体1 肺上皮细胞 细胞凋亡 炎症反应 

分 类 号：R446[医药卫生—诊断学] R563[医药卫生—临床医学]