摘 要: 目的 探讨高尔基体磷蛋白3(golgiosomal phosphoprotein 3,GOLPH3)调控JAK2/STAT3信号通路在镍精炼烟尘致人支气管上皮细胞(Beas-2B)炎性损伤中的作用。方法 采用不同浓度镍精炼烟尘(0.00、12.50、25.00、50.00 μg/ml)处理Beas-2B细胞24 h,使用CCK8法检测细胞存活率;采用蛋白免疫印迹(Western blot,WB)法检测GOLPH3及JAK2/STAT3信号通路蛋白JAK2、p-JAK2、STAT3、p-STAT3和炎性效应蛋白NLRP3、白细胞介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)表达水平。使用siRNA转染沉默GOLPH3,采用WB法检测GOLPH3、JAK2、p-JAK2、STAT3、p-STAT3、NLRP3、IL-18、TNF-α蛋白表达水平;使用JAK2/STAT3通路抑制剂AG490抑制该信号通路,采用WB法检测JAK2、p-JAK2、STAT3、p-STAT3及NLRP3、IL-18、TNF-α蛋白表达水平。结果 随着镍精炼烟尘染毒浓度增加,Beas-2B细胞存活率显著降低(P<0.05);GOLPH3、p-JAK2、p-STAT3、NLRP3、IL-18、TNF-α蛋白表达水平呈逐渐升高趋势(P<0.05);与si-对照组相比,si-GOLPH3组Beas-2B细胞中GOLPH3、p-JAK2、p-STAT3和NLRP3、IL-18、TNF-α表达均降低(P<0.05);与二甲基亚砜(dimethyl sulfoxide,DMSO)组相比较,AG490组Beas-2B细胞中p-JAK2、p-STAT3和NLRP3、IL-18、TNF-α表达均降低(P<0.05)。结论 镍精炼烟尘可通过激活GOLPH3调控JAK2/STAT3信号通路,诱导Beas-2B细胞发生炎性损伤。 |
关键词: 镍精炼烟尘 炎症 JAK2/STAT3 细胞毒性 |
中图分类号: R994.3
文献标识码: A
|
基金项目: 国家自然科学基金资助项目(编号:81973010) |
|
Role of Golgisome phosphoprotein 3 in inflammatory damage of human bronchial epithelial cells induced by nickel refining fumes |
ZHANG Dan,GAO Ying,JU Liyan,ZHANG Tong,CHEN Yangyang,YAO Wenxue,YANG Shikuan,LI Lina,WU Yonghui
|
School of Public Health,Harbin Medical University,Harbin,Heilongjiang 150081,China
|
Abstract: Objective To explore the role of Golgisome phosphoprotein 3(GOLPH3)in regulating the JAK2/STAT3 signaling pathway in inflammatory injury of human bronchial epithelial cells(Beas-2B)induced by nickel refining fumes. Methods The survival rate of Beas-2B cells was detected by CCK8 method after treating with different concentrations of nickel refining fumes(0.00,12.50,25.00 and 50.00 μg/ml);Western blot(WB)method was used to detect the expression levels of the GOLPH3,JAK2/STAT3 pathway proteins JAK2、p-JAK2、STAT3、p-STAT3 and the inflammation-related proteins NLRP3、IL-18 and TNF-α;silencing GOLPH3 with small interfering RNA(siRNA)transfection and WB method was used to detect the expression levels of GOLPH3、JAK2、p-JAK2、STAT3、p-STAT3、NLRP3、IL-18 and TNF-α;AG490 was used to inhibit JAK2/STAT3 signaling pathway and WB method was used to detect the expression levels of GOLPH3、JAK2、p-JAK2、STAT3、p-STAT3、NLRP3、IL-18 and TNF-α. Results The results showed that the survival rates of Beas-2B cells decreased significantly with the increase of nickel refining fumes exposure concentration(P<0.05);the protein expression levels of GOLPH3、p-JAK2、p-STAT3、NLRP3、IL-18 and TNF-α increased significantly(P<0.05);while compared with the si-control group,the protein expression levels of GOLPH3、p-JAK2、p-STAT3、NLRP3、IL-18 and TNF-α were decreased in si-GOLPH3 group(P<0.05),compared with the dimethyl sulfoxide(DMSO)group,the protein expression levels of GOLPH3、p-JAK2、p-STAT3、NLRP3、IL-18 and TNF-α were also decreased in AG490 group(P<0.05). Conclusion The results suggested that the nickel refining fumes could induce inflammatory injury of Beas-2B cells by activating GOLPH3 to regulate JAK2/STAT3 signaling pathway. |
Keywords: nickel refining fumes inflammation JAK2/STAT3 cytotoxicity |