CAJ | 학술논문

目的：观察血小板因子4(PF4)是否通过核因子κB (NF-κB)上调THP-1单核源性巨噬细胞基质金属蛋白酶9( MMP-9)表达。方法佛玻酯诱导THP-1细胞分化成巨噬细胞。巨噬细胞分别在NF-κB抑制剂( PDTC)缺乏或存在情况下与溶媒或PF4(100滋g/L)孵育一定时间,RT-PCR检测MMP-9 mRNA水平；同时,巨噬细胞与PF4(25~200滋g/L)单独或结合Toll样受体4(TLR4)阻断剂(抗体 HTA125,anti-TLR4)孵育,ELISA法检测NF-κB含量。结果 PF4较对照组上调巨噬细胞MMP-9 mRNA水平。而NF-κB抑制剂抑制PF4诱导的巨噬细胞MMP-9表达上调。 PF4呈浓度依赖性增加巨噬细胞的NF-κB含量,最大效应浓度为100滋g/L。 TLR4阻断剂逆转PF4诱导的巨噬细胞NF-κB含量增加。结论 PF4可能通过NF-κB上调巨噬细胞MMP-9的表达。 TLR4可能是PF4-NF-κB通路中NF-κB的上游信号分子。
목적：관찰혈소판인자4(PF4)시부통과핵인자κB (NF-κB)상조THP-1단핵원성거서세포기질금속단백매9( MMP-9)표체。방법불파지유도THP-1세포분화성거서세포。거서세포분별재NF-κB억제제( PDTC)결핍혹존재정황하여용매혹PF4(100자g/L)부육일정시간,RT-PCR검측MMP-9 mRNA수평；동시,거서세포여PF4(25~200자g/L)단독혹결합Toll양수체4(TLR4)조단제(항체 HTA125,anti-TLR4)부육,ELISA법검측NF-κB함량。결과 PF4교대조조상조거서세포MMP-9 mRNA수평。이NF-κB억제제억제PF4유도적거서세포MMP-9표체상조。 PF4정농도의뢰성증가거서세포적NF-κB함량,최대효응농도위100자g/L。 TLR4조단제역전PF4유도적거서세포NF-κB함량증가。결론 PF4가능통과NF-κB상조거서세포MMP-9적표체。 TLR4가능시PF4-NF-κB통로중NF-κB적상유신호분자。
Objective To investigate whether PF4 modulates the MMP-9 expression of macrophages via Nuclear fac-tor kappa B(NF-κB). Methods THP-1 monocytes were differentiated into monocyte-derived macrophages by phorbol 12-myristate 13-acetate (PMA). Macrophages were incubated with PF4 (100 μg/L) in the absence or presence of NF-κB inhibitor pyrrolidine dithiocarbamate ( PDTC) ,the MMP-9 expression of macrophages was determined by Reverse-transcrip-tion polymerase chain reaction (RT-PCR). Macrophages were incubated with PF4 (25-200 μg/L) or vehicle (PBS),NF-κB content in cultured supernatant of macrophages was measured by ELISA assay. To evaluate the intracellular signal trans-duction pathways,macrophages were pretreated for 30min with the TLR4 blocker(monoclonal antibody HTA125,anti-TLR4) before the addition of PF4,NF-κB content was measured. Results Macrophages that were untreated showed a relatively low MMP-9 mRNA level;treatment with PF4 increased MMP-9 expression. However,the high levels of MMP-9 expression induced by PF4 were significantly attenuated in the presence of NF-κB inhibitor. PF4 increased concentration of NF-κB in a dose dependent manner,with the strongest increase at 100ng/mL. The increased the concentration of NF-κB induced by PF4 was significantly reduced when treated with TLR4 blocker. Conclusion PF4 may up-regulate MMP-9 expression in mac-rophages via NF-κB. TLR4 may be upstream signaling molecules of NF-κB in PF4-NF-κB signaling pathway.