芹菜素对脂多糖诱导的炎性体活化调节的实验观察

目的 探讨芹菜素对脂多糖诱导的炎症中炎性体活化是否有调节作用,并初步研究其作用机制。方法 利用药物处理急性单核白血病细胞株THP-1,设置脂多糖处理组,脂多糖+25μmol/L芹菜素处理组,脂多糖+50 μmol/L芹菜素处理组及脂多糖+Z-VAD半胱氨酸蛋白酶（caspase）抑制剂]处理组,以二甲基亚砜(DMSO)处理作为对照组。通过酶联免疫吸附试验(ELISA)检测上清中白细胞介素1β(IL-1β)的含量;通过Western印迹检测IL-1β及caspase-1的剪切;通过报告基因方法检测芹菜素对炎症转录因子核因子(NF)κB活性的影响。结果 四甲基偶氮唑盐(MTT)比色法表明芹菜素对细胞未见明显毒性。在THP-1细胞中,芹菜素对于脂多糖诱导的炎性体活化产物IL-1β的产生具有显著的抑制作用上清中IL-1β浓度：对照组为(362±64) pg/ml,脂多糖组为(1549±320) pg/ml,脂多糖+25 μmol/L芹菜素组为(397±150) pg/ml,脂多糖+50 μmol/L芹菜素组为( 268±142) pg/ml,P＜0.05]。芹菜素能够显著抑制IL-1β前体(pm-IL-1β)以及炎性体成分caspase-1前体（pro-caspase-1）的成熟过程,并且能够抑制脂多糖诱导的NF-κB的活化（NF-κB活性相对荧光值：对照组为0.6±0.1,脂多糖组为32.7±0.8,脂多糖+25μmol/L芹菜素组为12.9±1.8,脂多糖+ 50 μmol/L芹菜素组为10.0±3.2,P＜0.05）。结论 芹菜素能够通过抑制caspase-1的成熟抑制脂多糖诱导的炎性体的活化。

Apigenin regulates lipopolysaccharides-induced activation of inflammasome

Objective To evaluate whether or not apigenin regulates the activation of inflammasome and elucidate its underlying mechanism. Methods Cultured THP-1 (acute monocytic leukemia cell line) cells were treated with dimethyl sulfoxide (DMSO) alone (control group), lipopolysaccharides (LPS), LPS plus apigenin (25/50 μmol/L) or LPS plus Z-VAD (a caspase inhibitor). The supernatant was harvested and the content of secreted interleukin (IL)-1β3 was determined by enzyme-linked immunosorbent assay (ELISA). The effects of apigenin on the cleavage of pro-IL-1β and pro-caspase-1 were determined by Western blot. And the effect of apigenin on the nuclear factor (NF) -κB activation was detected by reporter gene assay. ResultsMTT assays showed that the cytotoxicity of apigenin was low. Apigenin could sigrificantly inhibit the LPS-induced secretion of IL-1β in THP-1 cells. The concentration of IL-1β was (362 ±64) pg/ml in the control group, ( 1549 ±320) pg/ml in the LPS group, (397 ± 150) pg/ml in the LPS plus 25 μmol/L apigenin group and (268 ± 142) pg/ml in the LPS plus 50 μmol/L apigenin group (P ＜0. 05).The results of Western blot indicated that apigenin inhibited the maturation of pro-IL-1β and pro-caspase-1.It could also inhibit the LPS-induced activation of NF-κB. The value of relative light unit was 0. 6 ± 0. 1 in the control group, 32. 7 ±0. 8 in the LPS group, 12. 9 ± 1.8 in the LPS plus 25 μmol/L apigenin group and 10. 0 ± 3. 2 in the LPS plus 50 μmol/L apigenin group respectively (P ＜ 0. 05 ) . Conclusion Apigenin may inhibit the LPS-induced activation of inflammasome through an inhibited muturation of caspase-1.