目的:本研究拟探讨PDTC和姜黄素对抗β2GPI抗体诱导小鼠组织因子(Tissue factor,TF)表达的影响。方法:BALB/c小鼠先用PDTC[100 mg/(kg·d)]或/和姜黄素[50 mg/(kg·d)]预处理2 h,再进行腹腔注射anti-β2GPI(500μg/只),取小鼠腹腔巨噬细胞、双侧颈动脉和胸主动脉,超声裂解,收集细胞总RNA和蛋白,实时定量PCR(Real-time q PCR)检测细胞TF mRNA水平,TF活性试剂盒检测TF活性,Western blot方法检测NF-κB p65、磷酸化NF-κB p65、c-Jun和磷酸化pc-Jun的表达情况。结果:抗β2GPI抗体能够诱导小鼠腹腔巨噬细胞TF的表达及NF-κB p65和c-Jun/AP-1的磷酸化,与对照相比差异有统计学意义(P〈0.05 vs NR-Ig G);PDTC和姜黄素均能抑制抗β2GPI抗体诱导小鼠TF的表达以及NF-κB p65和cJun/AP-1的磷酸化效应(P〈0.05 vs anti-β2GPI),但是二者没有协同作用。结论:PDTC和姜黄素均能影响抗β2GPI抗体诱导小鼠TF的表达,表明其具有防治血栓形成的潜力。
Objective: To investigate whether PDTC or cureumin had effect on anti-β2GPI-indueed tissue factor (TF) expression in mice. Methods:BALB/c mice were pretreated with PDTC (100 mg/kg, once a day) by intraperitoneal injection (i. p. ) or/and cureumin (50 mg/kg,onee a day) by oral gavage for 3 consecutive clays at 2 h before 500 ttg of anti-[32GPI injections in subsequent experiments. Mouse peritoneal macrophages and aorta were collected, homogenized by sonication. The total RNA and protein were collected from each animal, TF expression was detected by Real-time quatitative PCR and TF activity kit. The phosphorylation of NF-KB 1365 and e-Jun/AP-1 was determined by Western blot. Results: Anti-β2GPI cloud significantly upregulate TF expression and phosphorylation of NF-KB 1365 and c-Jun/AP-1 in mouse peritoneal macrophages and aorta,compared with NR-IgG treated mice (P〈 0. 05 ). PDTC or/and curcumin could markedly attenuate anti-β2GPI-induced TF expression, also inhibit activation of NF-KB p65 and c- Jun/AP-1 in the aorta and peritoneal macrophages respectively ( P 〈 0. 05 ), but combination of two inhibitors had no synergistic effect. Conclusion:Both PDTC and curcUmin could affect the expression of TF induced by anti-β2GPI in mice, indicating that PDTC and curcumin has the potential to prevent thrombosis in APS.