δ阿片受体激动剂对LPS诱导的巨噬细胞凋亡及坏死的影响

Effect of delta-opioid receptor agonist on LPS induced apoptosis and necrosis of macrophages

目的研究δ阿片受体激动剂DADLE对LPS诱导的巨噬细胞凋亡和坏死的影响。方法用LPS(100μg/L)刺激大鼠腹腔巨噬细胞,在LPS后立即或4 h后加入DADLE(10-6M)。流式细胞仪和共聚焦成像检测巨噬细胞凋亡及坏死,ELISA法检测巨噬细胞细胞核NF-κB的活化水平,Western blot法检测p38 MAPK活化水平,ELISA法检测细胞培养上清TNF-α、IL-1β和HMGB1水平。结果 DADLE明显抑制了LPS诱导的巨噬细胞凋亡及坏死,降低了细胞培养上清中TNF-α、IL-1β和HMGB1的水平,而且对巨噬细胞NF-κB及p38 MAPK的活化均有抑制作用。结论 DADLE通过抑制NF-κB及p38 MAPK的活化,减少了LPS诱导的巨噬细胞凋亡、坏死及细胞因子的释放。

Objective To study the effect of deha-opioid receptor agonist （DADLE） on LPS induced apoptosis and necrosis of maerophages. Methods Peritoneal macrophages isolated from rats were challenged with LPS. DADLE at 104 M was administrated concurrently or 4h after LPS. Apoptosis and necrosis of macrophages were determined by flow cytometry analysis and confocal imaging. NF-KB activity in macrophages was determined by ELISA. Levels of activated p38 MAPK were measured by western blot. Levels of TNF-α,IL-1β and HMGB1 in culture supernatant were determined by ELISA. Results TNF-α,IL-1β and HMGB1 in supernatant were also suppressed by DADLE. LPS-induced apoptosis and necrosis of maerophages were significantly suppressed by DADLE, DADLE also inhibited the p38 MAPK and NF-KB pathways. Conclusion DADLE attenuates LPS induced apoptosis and necrosis of macrophages and cytokine release by suppressing the p38 MAPK and NF-KB pathways.