MKP-1/ERK通路参与雌二醇减轻中性粒细胞迁移的机制研究  

The study of mechanism of MKP-1/ERK pathway involved in estradiol reducing neutrophil migration

作　　者：金乾衡 徐磊 金光哲[1] 巨积辉 侯瑞兴 Jin Qianheng;Xu Lei;Jin Guangzhe;Ju Jihui;Hou Ruixing(Department of Hand Surgery,Suzhou Ruihua Orthopedic Hospital,Suzhou 215104,China;Suzhou Medicine College,Suzhou University,Suzhou 215123,China;Medical College,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]苏州瑞华骨科医院手外科,苏州215104 [2]苏州大学苏州医学院,苏州215123 [3]扬州大学医学院,扬州225009

出　　处：《中华手外科杂志》2023年第3期271-275,共5页Chinese Journal of Hand Surgery

基　　金：苏州市重点学科(SZXK202127);苏州市科技计划项目(SYSD2019054);苏州市科教兴卫项目(KJXW2019073)。

摘　　要：目的研究雌二醇对f-MLP诱导的体外中性粒细胞活化的影响,并探讨这种作用与ERK通路及其上游蛋白MKP-1的关系.方法将HL-60诱导分化成类中性粒细胞(dHL-60).用MTT实验检测不同浓度梯度的E2、f-MLP、PD98059或混合液对dHL-60的影响.通过Transwell实验和超氧化物检测E2、PD98059对f-MLP诱导的dHL-60迁移和超氧化的抑制作用.Western blot检测ERK、p-ERK、MKP-1的表达水平.结果E2、f-MLP、PD98059单独或联合使用对dHL-60细胞的存活率没有影响.f-MLP能显著诱导中性粒细胞迁移和过度氧化.生理、药理学剂量的E2和PD98059可以有效抑制f-MLP的这种作用.Western blot检测结果显示f-MLP诱导ERK磷酸化,而E2和PD98059有效抑制f-MLP的这种作用.另外,雌二醇能有效地增加MKP-1的表达.结论雌二醇可能通过增加MKP-1蛋白降低ERK磷酸化,从而抑制f-MLP诱导的体外中性粒细胞迁移及超氧化.Objective To study the effect of estradiol on the activation of neutrophils induced by f-MLP in vitro,and to explore the relationship between this effect and ERK pathway and its upstream protein MKP-1.Methods HL-60 was induced to differentiate into neutrophil-like cells(dHL-60).The effects of E2,f-MLP,PD98059 or mixture of different concentration gradients on dHL-60 were detected by MTT test.The inhibitory effect of E2 and PD98059 on f-MLP induced dHL-60 migration and superoxide was detected by Transwell experiment and superoxide test.The expression levels of ERK,p-ERK and MKP-1 were detected by Western blot.Results E2,f-MLP and PD98059 alone or in combination had no effect on the survival rate of dHL-60 cells.The f-MLP can significantly induce neutrophil migration and over oxidation.The physiological and pharmacological doses of E2 and PD98059 can effectively inhibit this effect of f-MLP.The results of Western blot showed that f-MLP induced ERK phosphorylation,and E2 and PD98059 effectively inhibited this effect of f-MLP.In addition,estradiol can effectively increase the expression of MKP-1.Conclusion Estradiol may decrease ERK phosphorylation by increasing MKP-1 protein,thereby inhibiting f-MLP induced neutrophil migration and superoxidation in vitro.

关 键 词：雌二醇 超氧化物类 中性粒细胞迁移 MKP-1/ERK 类风湿性关节炎 

分 类 号：R96[医药卫生—药理学]