| 乐翊飞,汪俊成,唐利华,等.天名精根提取物调控TLRs信号抑制RAW264.7细胞炎症的研究[J].浙江中医药大学学报,2019,43(4):360-367. |
| 天名精根提取物调控TLRs信号抑制RAW264.7细胞炎症的研究 |
| Root Extracts from Carpesium Abrotanoides Inhibits RAW264.7 Cell Inflammation by Regulating TLRs Signaling Pathway |
| DOI：10.16466/j.issn1005-5509.2019.04.016 |
| 中文关键词: 天名精 RAW264.7 脂多糖 金黄色葡萄球菌 中药提取物 炎症 |
| 英文关键词: Carpesium abrotanoides Linn. RAW264.7 lipopolysaccharide Staphylococcus aureus extract of traditional Chinese medicine inflammation |
| 基金项目:浙江省大学生科技创新活动计划(新苗人才计划)项目(772214F00802) |
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| 中文摘要: |
| [目的]阐明天名精根提取物对RAW264.7细胞炎症反应的影响及其机制。[方法]采集野生天名精全株，处理得天名精根粉末，制备天名精根乙醇和石油醚提取物，并以高效液相色谱法(high performance liquid chromatography，HPLC)检测其中主要成分；采用脂多糖(lipopolysaccharide，LPS)、灭活的金黄色葡萄球菌(heat-inactivated preparations of Staphylococcus aureus pathogens，SAC)刺激RAW264.7细胞，构建体外炎症模型。以MTT法检测细胞增殖，qPCR检测白细胞介素-1β(interleukin-1β，IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、Toll样受体-2(Toll-like receptor-2，TLR-2)、TLR-4、髓样分化因子88(myeloid differentiation primary response 88，MyD88)、核转录因子-κB(nuclear factor-κB，NF-κB)mRNA表达，Western blot检测NF-κB p65蛋白表达。[结果]与正常对照组比较，LPS或SAC刺激后RAW264.7细胞IL-1β、IL-6、TNF-α mRNA表达增加(P＜0.01)，NF-κB蛋白与mRNA表达增加(P＜0.01)；LPS刺激后TLR-4、MyD88 mRNA表达增加(P＜0.01，P＜0.05)，SAC刺激后TLR-2、MyD88 mRNA表达增加(P＜0.01)。天名精根提取物干预后，与LPS组或SAC组比较，RAW264.7细胞IL-1β、IL-6、TNF-α mRNA表达减低(P＜0.01)，同时NF-κB mRNA和NF-κB p65蛋白表达也减低(P＜0.01)；TLR-4和MyD88 mRNA表达明显低于LPS组(P＜0.01，P＜0.05)，TLR-2和MyD88 mRNA表达表达明显低于SAC组(P＜0.01，P＜0.05)。[结论]天名精根提取物可以抑制由LPS和灭活的金黄色葡萄球菌引起的RAW264.7细胞炎症反应，其机制可能与抑制TLRs-NF-κB信号通路有关。 |
| 英文摘要: |
| [Objective] To elucidate the effect and mechanism of the root extract of Carpesium abrotanoides Linn.(CA) on the inflammatory response of RAW264.7 cells.[Methods] The effective components of CA were separated, and the constituents were detected by high performance liquid chromatography(HPLC).In vitro inflammatory model was constructed by using lipopolysaccharide(LPS) and heat-inactivated preparations of Staphylococcus aureus pathogens(SAC) to stimulate RAW264.7 cells. MTT assay was used for cell proliferation detection; mRNA expression of interleukin-1β(IL-1β), IL-6, TNF-α, Toll-like receptor-2(TLR-2), TLR-4, myeloid differentiation primary response 88 (MyD88), nuclear factor-κB(NF-κB) was detected by qPCR; protein expression of NF-κB p65 was detected by Western blot. [Results] Compared with normal control group, the expression of IL-1β, IL-6, TNF-α mRNA and NF-κB protein and mRNA increased(P＜0.01), after treated with LPS or SAC. After treated with LPS, the expression of TLR-4, MyD88 mRNA increased (P＜0.01，P＜0.05); and the expression of TLR-2, MyD88 mRNA increased after treated with SAC(P＜0.01). Compared with LPS group or SAC group, the expression of IL-1β, IL-6, TNF-α mRNA and NF-κB protein and mRNA decreased after treated with the root extract of CA(P＜0.01).The expression of TLR-4 and MyD88 mRNA were lower than LPS group(P＜0.01，P＜0.05), and the expression of TLR-2 and MyD88 mRNA were lower than SAC group(P＜0.01，P＜0.05). [Conclusion]The root extract of CA can inhibit the in vitro inflammatory response of RAW264.7 cells caused by LPS and SAC, which may be due to inhibition of TLRs-NF-κB signaling pathway. |
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