Effects of mTOR activation on anti-β2GPI/β2GPI-stimulated tissue factor expression in THP-1 cells
10.13602/j.cnki.jcls.2017.03.17
- VernacularTitle:mTOR活化对抗β2GPI抗体/β2GPI复合物诱导THP-1细胞组织因子表达的影响
- Author:
Longfei XIA
;
Hongmei JU
;
Hong ZHOU
;
Ting WANG
- Keywords:
antiphospholipid syndrome;
mammalian target of rapamycin(mTOR);
anti-β2GPI/β2GPI complex;
tissue factor;
rapamycin
- From:
Chinese Journal of Clinical Laboratory Science
2017;35(3):215-220
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of mammalian target of rapamycin(mTOR) in the expression of tissue factor(TF) from THP-1 cells induced by β2GPI/anti-β2GPIcomplex.Methods The THP-1 cells were treated with both β2GPI/anti-β2GPI and β2GPI/IgG-APS(β2GPI/IgG from APS patients) complexes.Rapamycin(100 nmol/L),the mTOR inhibitor,was used to exert the intervention experiment.The total RNA and proteins of the THP-1 cells were collected for detection.The mRNA expression level and activity of TF in THP-1 cells were detected by real-time quatitative PCR(RT-qPCR) and TF activity kit respectively.western blotwas used to determine the levels of mTOR and phosphorylated-mTOR(p-mTOR),and p38,p-p38,ERK1/2,p-ERK1/2,JNK,p-JNK,NF-κB p65 and p-NF-κB p65 in THP-1 cells were determined simultaneously.Results Both β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes chould significantly upregulate the mRNA expression and activity of TF,and the phosphorylation levels of mTOR in THP-1 cells(P ＜ 0.05).Rapamycin markedly attenuated the mRNA expression and activity of TF and mTOR phosphorylation induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P ＜ 0.05),and also inhibited the phosphorylation levels of p38,ERK1/2 and NF-κB p65 in THP-1 cells induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P ＜ 0.05),but did not showed effects on the phosphorylation of c-Jun NH2-terminal protein kinase (JNK) (P ＞ 0.05).Conclusion mTOR could be activated by β2GPI/antiβ2GPI complexes in THP-1 cells and play a crucial role for β2GPI/anti-β2GPI-induced TF expression in THP-1 cells.
