Sinomenine regulates CD14/TLR4, JAK2/STAT3 pathway and calcium signal via α7nAChR to inhibit inflammation in LPS stimulated macrophages
Sinomenine regulates CD14/TLR4, JAK2/STAT3 pathway and calcium signal via α7nAChR to inhibit inflammation in LPS stimulated macrophages
复制标题
青藤碱通过 α7nAChR 调节 CD14/TLR4、JAK2/STAT3 通路和钙信号，抑制 LPS 刺激的巨噬细胞炎症
DOI：
10.1080/08923973.2019.1568451
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发表时间：
2019
影响因子：
3.3
通讯作者：
Yan Dong
中科院分区：
文献类型：
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作者：
Rui-li Zhu;Ying-kun Zhi;Lang Yi;Jin-fang Luo;Jing Li;Sha-sha Bai;Liang Liu;Pei-xun Wang;Hua Zhou;Yan Dong
目的：探讨青藤碱(SIN)通过α7烟碱型乙酰胆碱受体(α7nAChR)抑制脂多糖诱导的巨噬细胞炎症的细胞机制。用α7nAChR的选择性拮抗剂α银环蛇毒素(Btx)阻断α7nAChR。用AG490抑制JAK2的激活。采用双抗体夹心ELISA法检测血清肿瘤坏死因子-α和单核细胞趋化蛋白-1水平。免疫印迹法检测MIF、MMP9、CD14、TLR4、STAT3、p-STAT3的表达。结果：SIN可抑制RAW 264.7细胞产生肿瘤坏死因子-α、单核细胞趋化蛋白-1、巨噬细胞间黏附分子和基质金属蛋白酶-9，减少CD14和TLR4的表达，抑制细胞内钙离子的释放。JAK特异性抑制剂AG490可减弱SIN对肿瘤坏死因子-α的抑制作用。SIN使STAT3的磷酸化水平升高。α7nAChR拮抗剂可减弱SIN的上述作用。结论：SIN可通过α7nAChR降低巨噬细胞CD14TLR4表达和细胞内游离钙水平，激活JAK2/STAT3通路，从而抑制炎症反应。
Objective:To investigate the cellular mechanism that sinomenine (SIN) inhibits inflammation in macrophages induced by LPS through α7 nicotinic acetylcholine receptor (α7nAChR).Materials and methods:RAW264.7 cells were stimulated with LPS and treated by SIN or nicotine (Nic). A selective antagonist of α7nAChR, α-bungarotoxin (BTX) was used to block α7nAChR. AG490 was used to inhibit JAK2 activation. ELISA was performed to detect the levels of TNF-α and MCP-1. Western blotting was used to analyze the expression of MIF, MMP-9, CD14, TLR4, STAT3 and p-STAT3. Intracellular-free calcium level was measured by Fluorescent probe fluo-3/AMResults:SIN inhibited the production of TNF-α, MCP-1, MIF, and MMP-9, decreased the expression of CD14 and TLR4, and inhibited the release of intracellular-free calcium from intracellular stores in RAW 264.7 cells stimulated by LPS. JAK-specific inhibitor AG490 attenuated the inhibitory effect of SIN on TNF-α. SIN increased the phosphorylation of STAT3. And the above effects of SIN were attenuated by antagonist of α7nAChR.Conclusions:SIN can decrease the expression of CD14/TLR4 and intracellular free calcium level, activate JAK2/STAT3 pathway to inhibit inflammatory response through α7nAChR in macrophages.