Isoforskolin Inhibits Lipopolysaccharide Activation of the TLR4/MyD-88/NF-κB Pathway in Small Airway Epithelial Cells
Isoforskolin Inhibits Lipopolysaccharide Activation of the TLR4/MyD-88/NF-κB Pathway in Small Airway Epithelial Cells
复制标题
异毛喉素抑制小气道上皮细胞中 TLR4/MyD-88/NF-κB 通路的脂多糖激活
DOI：
10.29011/2575-789x.000151
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Weimin Yang
中科院分区：
文献类型：
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作者：
Xiaohua Du;Chen Chen;Yin Huang;Peihua Peng Shibo Sun XiulinYe Ling Li;Zelan Dai Xiaoyue Xiao;Zhiying Weng;Weimin Yang
**摘要**。目的：异佛司可林（ISOF）分离自原产于中国云南的毛喉鞘蕊花植物，其生物学效应备受关注。已有研究表明它可减轻脂多糖（LPS）诱导的大鼠急性肺损伤（ALI）的炎症反应。在本研究中，我们假设ISOF可抑制LPS在人小气道上皮细胞（BEAS - 2B）中激活的一条主要信号通路：Toll样受体4（TLR4）、髓样分化因子88（MyD88）和核因子κB（NF - κB）信号级联反应。方法：在给予LPS（2 μg/mL）刺激前，用ISOF（1.0μM）、佛司可林（FSK，1.0μM）、罗氟司特（RF，1.0μM）和地塞米松（DEX，50μM）对BEAS - 2B细胞进行预处理。采用蛋白质免疫印迹法和实时荧光定量PCR检测BEAS - 2B细胞中TLR4、MyD88和NF - κB的蛋白及mRNA水平。使用MTT法检测ISOF和LPS对细胞活力的影响。结果：我们的研究结果显示，LPS可增加BEAS - 2B细胞中TLR4、MyD88和NF - κB的蛋白及mRNA水平。LPS处理后，BEAS - 2B细胞中TLR4、MyD88、NF - κB的蛋白及mRNA水平显著下降。ISOF预处理可减弱LPS的作用。此外，在6小时时，ISOF（剂量< 5.0 μM）和LPS（剂量< 20 μg/mL）对BEAS - 2B细胞活力无影响。结论：这些结果表明，ISOF通过下调BEAS - 2B细胞中TLR4、MyD88和NF - κB的mRNA和蛋白水平，抑制由LPS激活的TLR4/MyD88/NF - κB信号通路。
Abstract.Object: Isoforskolin (ISOF) is isolated from the plant Coleus forskohlii, native to Yunnan China, which has attracted much attention for its biological effects. It has been demonstrated to attenuate inflammation of rat ALI induced by Lipopolysaccharide (LPS). In this study, we hypothesize that ISOF inhibits a main pathway activated by LPS in human small airway epithelial cells (BEAS-2B): cascade of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and nuclear factor kappa-B (NF-κB) signaling..Methods: BEAS-2B cells were pretreated with ISOF (1.0μM), forskolin (FSK, 1.0μM), roflumilast (RF, 1.0μM) and dexamethasone (DEX, 50μM) prior to LPS (2 μg/mL). Protein and mRNA levels of TLR4, MyD88, and NF-κB in BEAS-2B were determined using Western blot and Real-time quantitative PCR. Effects of ISOF and LPS on cell viability were tested by using MTT method..Results: Our results showed that LPS augmented the protein and mRNA levels of TLR4, MyD88, and NF-κB in BEAS-2B cells. Following LPS treatment, protein and mRNA levels of TLR4, MyD88, NF-κB in BEAS-2B decreased significantly. Pretreatment with ISOF attenuated the effects of LPS. Moreover, ISOF (with dose < 5.0 μM) and LPS (with dose < 20 μg/mL) has no effect on cell viability of BEAS-2B at 6h..Conclusions: These results have shown that ISOF inhibits TLR4/MyD88/NF-κB signal pathway, which is activated by LPS, through down-regulation of mRNA and protein levels of TLR4, MyD88 and NF-κB in BEAS-2B.