摘要
目的探讨甘草查尔酮A(Lico A)对脂多糖(LPS)诱导的人急性单核细胞白血病细胞株(THP-1)巨噬细胞相关炎症因子表达的影响。方法用100μg/L佛波酯(PMA)诱导THP-1细胞48 h,使其分化为巨噬细胞后,分为空白组、LPS组和LPS+不同浓度Lico A组(20、10、5 mg/L)。用酶联免疫吸附法检测细胞培养液中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的含量,实时定量PCR检测Toll样受体4(TLR-4)和核因子κB(NF-κB)m NRA水平,采用Western blot检测TLR-4、NF-κB、IκB激酶(IKKα)、磷酸化IKB-α(p-IKB-α)、环氧合酶2(COX-2)和一氧化氮合酶(i NOS)蛋白表达水平。结果 LPS诱导THP-1巨噬细胞后,IL-1β、IL-6、TNF-α水平升高,Lico A可降低LPS诱导引起的IL-1β、IL-6和TNF-α表达水平升高。LPS刺激后TLR-4 m NRA及蛋白表达增加,NF-κB活化,Lico A可拮抗以上作用,阻止NF-κB活化。结论 Lico A可通过TLR-4/NF-κB通路抑制LPS诱导的THP-1巨噬细胞炎性反应。
Aim To investigate the effects of licochalcone A(Lico A) on the expression of inflammatory cytokines in macrophages of human acute monocytic leukemia cell line(THP-1) induced by lipopolysaccharide(LPS) in vitro.Methods 100 μg/L phorbol ester(PMA) was used to induce THP-1 cells for 48 hours,after the differentiation of THP-1 cells,which became macrophage,and cells were divided into control group,LPS group and LPS combined with concentration(20,10,5 mg/L) of Lico A. The levels of interleukin-1 beta(IL-1β),interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) were detected by ELISA. The levels of Toll like receptor-4(TLR-4) and nuclear factor kappa B(NF-κB) mRNAs were tested by real-time PCR. The levels of TLR-4,NF-κB,I kappa B kinase alpha(IKKα),phosphorylated IKB-alpha(p-IKB-α),cyclooxygenase-2(COX-2) and isoform of nitric oxide synthase(i NOS) proteins were examined by Western blot. Results The expression levels of TNF-α,IL-1 and IL-6 were up-regulated in the v macrophages after stimulated by LPS,Lico A could reduce the elevated expression levels of TNF-α,IL-1 and IL-6 induced by LPS. The expression of TLR-4 significantly increased after stimulated by LPS and NF-κB was activated. Lico A could reverse the above changes and prevent the activation of NF-κB. Conclusion Lico A could inhibit LPS-induced inflammatory response in THP-1 macrophages via TLR-4/NF-κB pathway.
出处
《中国动脉硬化杂志》
CAS
北大核心
2017年第12期1212-1218,共7页
Chinese Journal of Arteriosclerosis
基金
广西中药药效研究重点实验室开放项目(14-045-12)