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白术黄芪汤乙酸乙酯提取物对巨噬细胞NLRP3炎症小体的作用研究
Effects of Bai-Zhu Huang-Qi Decoction Extract on NLRP3 Inflammasome in Macrophages
  
DOI:
中文关键词:  关键词:炎症小体  白术黄芪汤乙酸乙酯提取物  NLRP3  caspase-1  IL-1β
英文关键词:Keywords: Inflammasome, Bai-Zhu Huang-Qi decoction ethyl acetate extract, NLRP3, caspase-1, IL-1β
基金项目:
作者单位
沈佳雯1 樊丹平2 邱雪梅1 吕爱平3 何小鹃2 耿 耘 1.西南交通大学生命科学与工程学院 成都 610031 2.中国中医科学院中医临床基础医学研究所 北京 100700 3.香港浸会大学中医药学院 香港 999077 
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中文摘要:
      摘 要:目的:观察白术黄芪汤乙酸乙酯提取物对巨噬细胞NOD样受体-3(NLRP3)炎症小体的影响。方法:用佛波酯(PMA)(10 ng?mL-1)刺激U937细胞48 h,诱导其成为巨噬细胞,观察不同剂量白术黄芪汤乙酸乙酯提取物(0,3.125,6.25,12.5,25,50,100 μg?mL-1)对细胞活力的作用,并选择合适的浓度。采用实时荧光定量(RT-PCR)和免疫印迹法(Western blot)测定细胞中NLRP3,半胱天冬酶-1(caspase-1)的含量,采用酶联免疫吸附测定(ELISA)检测细胞上清液中白细胞介素(IL)-1β 的含量。结果:细胞计数试剂盒(CCK-8)实验表明,当药物浓度低于25 μg?mL-1时,对细胞活力没有影响,当药物浓度高于50 μg?mL-1时,对细胞活力有抑制作用(P<0.05),选择25 μg?mL-1浓度进行后续实验。与空白组相比,LPS组细胞中NLRP3,caspase-1表达明显增加(P<0.01),细胞培养上清中的IL-1β浓度也明显增高(P<0.01)。白术黄芪汤乙酸乙酯提取物作用后,NLRP3,caspase-1,IL-1β表达均明显下降(P<0.05)。结论:白术黄芪汤乙酸乙酯提取物对LPS刺激的巨噬细胞NLRP3炎症小体有抑制作用。
英文摘要:
      Abstract: This study was aimed to observe the effect of Bai-Zhu Huang-Qi (BZHQ) decoction ethyl acetate extract on NOD like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome in macrophages. The U937 cells were pre-treated with phorbol 12-myristate 13-acetate (PMA, 10 ng?mL-1) for 48 hours to induce macrophages. Effects on cell viability by different doses of BZHQ decoction ethyl acetate extract (0, 3.125, 6.25, 12.5, 25, 50, 100 μg?mL-1) were observed to select the appropriate concentration. Contents of NLRP3 and caspase-1 in cells were detected by real-time PCR and western blot. The concentration of interleukin-1β (IL-1β) in cell supernatant was detected by enzyme linked immunosorbent assay (ELISA). The cell counting kit-8 (CCK-8) assay showed that when the drug concentration was lower than 25 μg?mL-1, there was no impact on cell viability; when the drug concentration was higher than 50 μg?mL-1, there was inhibition on cell viability (P < 0.05). The concentration of 25 μg?mL-1 was used to conduct the following experiment. Compared to the blank group, the expression of NLRP3 and caspase-1 in cells of the LPS group were significantly increased (P < 0.01). The concentration of IL-1β in cell supernatant was also significantly increased (P < 0.01). After treated with BZHQ decoction ethyl acetate extract, levels of NLRP3, caspase-1 and IL-1β were significantly decreased (P < 0.05). It was concluded that BZHQ decoction ethyl acetate extract can inhibit the production of NLRP3 inflammasome in LPS-stimulated macrophages.
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