摘要
目的探讨阿司匹林(aspirin)影响脂多糖(LPS)致猪肺泡巨噬细胞(AMs)环氧合酶-2(COX-2) mRNA转录及前列腺素E2(PGE2)表达的作用机制。方法AMs经阿司匹林单独或联合应用Calphostin C及过氧钒酸钠(POV)预处理后,分别以LPS刺激,RT-PCR方法测定AMs胞浆COX-2 mR-NA转录水平,ELISA方法测定AMs细胞培养上清PGE2表达水平。结果LPS(10μg/mL)刺激后,AMs胞浆COX-2 mRNA在1h即可检测到(0.70±0.16),并于4h达到峰值(1.86±0.40);AMs细胞培养上清PGE2水平在0.5h起开始上升,峰值出现在LPS刺激后12h左右。3mM阿司匹林可抑制LPS引起的AMs胞浆COX-2mRNA转录及细胞培养上清PGE2表达水平。蛋白酪氨酸磷酸酶(PTP)抑制剂POV可增强LPS刺激后AMsCOX-2 mRNA及PGE2表达水平,阿司匹林能减低该效应;与之相反,蛋白激酶C(PKC)抑制剂Calphostin C可减低LPS刺激后AMsCOX-2 mRNA及PGE2表达水平,阿司匹林能增强Calphostin C的作用。结论阿司匹林可能通过影响猪AMs蛋白激酶-磷酸酶系统平衡,抑制LPS致AMsIκBα磷酸化降解及NF-κB活化,进一步影响AMsCOX-2 mRNA表达及PGE2合成水平,减轻LPS引发急性肺损伤(ALI)炎性反应程度。
[Objective] To invdstigate the mechanism of Aspirin on the transcription of Cyclooxygenase-2 （COX- 2） mRNA and the expression of Prostaglandin E2 （PGE,） induced with Lipopolysaeeharide （LPS） in porcine alveolar maerophages （AMs）. [Methods] LPS was used to stimulate the AMs with or without the pretreatment of Aspirin （ASA）, Calphostin C （CAL） and Peroxovanadium（POV）, then the transcription of COX-2 mRNA and the expression of PGE2 was assayed by RT-PCR and ELISA respectively. [Results] The transcription of COX-2 mRNA in AMs increased at 1 hour after being stimulated with LPS（10μg/mL） and get peak value at 4 hours, while the expression of PGE2 in supematant of AMs increased at 30 minutes after being stimulated with LPS（10μg/mL） and get peak value at 12 hours later after the treatment of LPS. Aspirin（3 mM） can inhibit the transcription of COX-2 mRNA and the expression of PGE2 significantly in AMs induced with LPS（10μg/mL）. Aspirin can weaken the up-regulation effects of protein Tyrosine Phosphatase （FFP） inhibitor POV（20μM） on the transcription of COX-2 mRNA and the expression of PGE2 induced with LPS in AMs. On the contrary, Aspirin can enhance the down-regulation effects of the anspecific protein kinase C（PKC） inhibitor Calphostin C（0.5 μM） on the transcription of COX-2 mRNA and the expression of PGE2 indtlced by LPS in AMs respectively. [ Conclusions] The results in this paper and our previous study suggests that Aspirin can interfere the protein kinase and protein phosphatase balance system and NF-κB pathway to regulate the expression of PGE2 and inflammatory response of ALl induced with LPS in porcine alveolar macrophages.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第13期1804-1809,共6页
China Journal of Modern Medicine
基金
国家自然科学基金项目资助(No:39800064)
