Naringenin prevents TNF-α-induced gut-vascular barrier disruption associated with inhibiting the NF-κB-mediated MLCK/p-MLC and NLRP3 pathways.
Naringenin prevents TNF-α-induced gut-vascular barrier disruption associated with inhibiting the NF-κB-mediated MLCK/p-MLC and NLRP3 pathways.
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DOI：
10.1039/d1fo00155h
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发表时间：
2021-03
期刊：
影响因子：
6.1
通讯作者：
Jia Zhong;Ruyang Yu;Qilyu Zhou;Ping Liu;Zhongjie Liu;Yifei Bian
中科院分区：
文献类型：
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作者：
Jia Zhong;Ruyang Yu;Qilyu Zhou;Ping Liu;Zhongjie Liu;Yifei Bian
微血管内皮精确地调节分子穿过肠血管屏障（GVB）的通道，其在肠道免疫中起重要作用。据报道，柚皮素对几种肠道疾病具有治疗潜力。然而，柚皮素对GVB破坏的影响很少被研究。本研究旨在探讨柚皮素对GVB功能的影响及其可能机制。本研究用50 ng/mL肿瘤坏死因子-α（TNF-α）诱导GVB对大鼠小肠微血管内皮细胞（RIMVEC）的体外破坏作用。通过伊文思蓝（EB）-白蛋白流出试验和跨内皮电阻（TER）测定体外GVB的完整性。同时采用酶联免疫吸附试验（ELISA）、实时定量聚合酶链反应（RT-qPCR）、western blot和免疫荧光法检测细胞紧密连接蛋白及其相关的NF-κB、MLCK/p-MLC和NLRP 3通路的表达。结果表明，柚皮素（100 μM）抑制TNF-α诱导的白细胞介素（IL）-6分泌过多，减轻GVB破坏，并减轻紧密连接蛋白表达模式的变化。柚皮素抑制MLCK/p-MLC系统和TLR 4/NF-κB/NLRP 3通路的GVB破坏相关激活。此外，柚皮素在减少TNF-α诱导的NLRP 3、p-MLC活化和继发性GVB破坏方面显示出与NF-κB抑制剂Bay 11-7082相似的作用。结果表明，柚皮素通过维持紧密连接蛋白模式，部分抑制NF-κ B介导的MLCK/p-MLC和NLRP 3通路的活化，从而明显减轻TNF-α诱导的体外GVB破坏。
The microvasculature endothelium accurately regulates the passage of molecules across the gut-vascular barrier (GVB), which plays an essential role in intestinal immunity. Naringenin is reported to have therapeutic potential against several intestinal disorders. However, the effect of naringenin on GVB disruption has been rarely studied. This study aims to investigate the effect of naringenin on GVB function and the potential mechanism. In the present study, the in vitro GVB disruption of rat intestinal microvascular endothelial cells (RIMVEC) was induced by 50 ng mL-1 of tumor necrosis factor-α (TNF-α). The integrity of the in vitro GVB was determined by Evans blue (EB)-albumin efflux assay and trans-endothelial electrical resistance (TER). Meanwhile, the expression of tight junction proteins and the related NF-κB, MLCK/p-MLC and NLRP3 pathways were determined using enzyme-linked immunosorbent assay (ELISA), real-time quantitative polymerase chain reaction (RT-qPCR), western blot analysis and immunofluorescence. The results show that naringenin (100 μM) inhibits TNF-α-induced interleukin (IL)-6 hypersecretion, alleviates GVB disruption and mitigates the change in the tight junction protein expression pattern. Naringenin inhibits the GVB-disruption-associated activation of the MLCK/p-MLC system and TLR4/NF-κB/NLRP3 pathways. Furthermore, naringenin shows a similar effect to that of NF-κB inhibitor Bay 11-7082 in reducing the TNF-α-induced activation of NLRP3, p-MLC and secondary GVB disruption. The results suggest that naringenin evidently alleviates TNF-α-induced in vitro GVB disruption via the maintenance of a tight junction protein pattern, partly with the inhibition of the NF-κB-mediated MLCK/p-MLC and NLRP3 pathway activation.