目的 基于核因子κB抑制物激酶β（IKKβ）/核转录因子-κB（NF-κB）信号通路探讨藏红花素减轻高糖诱导的肾小球内皮细胞损伤的作用机制。 方法 以0、12.5、25、50、75 μmol/L藏红花素处理高糖诱导的人肾小球内皮细胞，通过CCK-8实验筛选藏红花素最佳作用浓度。肾小球内皮细胞随机分为正常组、渗透压对照组、高糖组、藏红花素组、佛波酯（PMA）组、藏红花素+PMA组，除正常组、渗透压对照组外其余各组以高糖诱导，与此同时用藏红花素和IKKβ/NF-κB信号通路激活剂PMA分组处理48 h。通过免疫荧光染色观察细胞骨架；通过CCK-8实验、Transwell实验、原位末端标记法（TUNEL）染色分别检测细胞活性、通透性与凋亡；测定细胞氧化应激与炎症相关因子水平；免疫印迹（WB）检测凋亡、紧密连接与IKKβ/NF-κB信号通路蛋白表达。 结果 与正常组、渗透压对照组相比，高糖组肾小球内皮细胞活性、超氧化物歧化酶（SOD）与谷胱甘肽过氧化物酶（GSH-Px）活性、occludin蛋白相对表达、闭锁蛋白1（ZO-1）与Bcl-2蛋白相对表达降低（P < 0.05），通透性、凋亡率、丙二醛（MDA）含量、白细胞介素（IL）-6与IL-1β水平、Bax蛋白相对表达、p-IKKβ/IKKβ、p-IκB-α/IκB-α、p-NF-κB p65/NF-κB p65升高（P < 0.05）。与高糖组相比，藏红花素组肾小球内皮细胞活性、SOD与GSH-Px活性、occludin蛋白相对表达、ZO-1与Bcl-2蛋白相对表达升高（P < 0.05），通透性、凋亡率、MDA含量、IL-6与IL-1β水平、Bax蛋白相对表达、p-IKKβ/IKKβ、p-IκB-α/IκB-α、p-NF-κB p65/NF-κB p65降低（P < 0.05）；PMA组各指标变化趋势与藏红花素组相反（P < 0.05）。与藏红花素组相比，藏红花素+PMA组肾小球内皮细胞活性、SOD与GSH-Px活性、occludin蛋白相对表达、ZO-1与Bcl-2蛋白相对表达降低（P < 0.05），通透性、凋亡率、MDA含量、IL-6与IL-1β水平、Bax蛋白相对表达、p-IKKβ/IKKβ、p-IκB-α/IκB-α、p-NF-κB p65/NF-κB p65升高（P < 0.05）。 结论 藏红花素通过抑制IKKβ/NF-κB信号通路激活减轻高糖诱导的肾小球内皮细胞损伤。

Objective To investigate the mechanism by which crocin alleviates high glucose-induced glomerular endothelial cell damage based on the inhibitor of nuclear factor kappa B kinase β （IKKβ）/nuclear transcription factor-κB （NF-κB） signaling pathway. Methods High glucose-induced human glomerular endothelial cells were treated with crocin at concentrations of 0， 12.5， 25， 50， and 75 μmol/L， and the optimal concentration of crocin was screened via the CCK-8 experiment. Glomerular endothelial cells were randomly assigned to the normal group， osmotic pressure control group， high glucose group， crocin group， phorbol ester （PMA） group， and crocin + PMA group. Except for the normal group and osmotic pressure control group， all the other groups were induced with high glucose and treated with crocin and the IKKβ/NF-κB signaling pathway activator PMA according to different groups for 48 hours. Immunofluorescence staining was employed to observe the cytoskeleton of the cells. The CCK-8 experiment， Transwell experiment， and TdT-mediated dUTP nick end labeling （TUNEL） staining were carried out to detect cell viability， permeability， and apoptosis， respectively. The oxidative stress and inflammation-related factors in the cells were measured. Western blot （WB） was utilized to detect the expression of proteins related to apoptosis， tight junctions， and the IKKβ/NF-κB signaling pathway. Results Compared with the normal group and the osmotic pressure control group， the high-glucose group exhibited a decrease in the activity of glomerular endothelial cells， the activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px）， the relative expression of occludin protein， and the relative expressions of zonula occludens-1 （ZO-1） and Bcl-2 protein （P < 0.05）. Meanwhile， it showed an increase in permeability， apoptosis rate， malondialdehyde （MDA） content， interleukin （IL）-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. Compared with the high-glucose group， the crocin group showed an increase in the activity of glomerular endothelial cells， the activities of SOD and GSH-Px， the relative expression of occludin protein， and the relative expressions of ZO-1 and Bcl-2 protein （P < 0.05）. In addition， it had a decrease in permeability， apoptosis rate， MDA content， IL-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. The trend of changes in various indicators in the PMA group was opposite to that in the crocin group （P < 0.05）. Compared with the crocin group， the crocin + PMA group showed a decrease in the activity of glomerular endothelial cells， the activities of SOD and GSH-Px， the relative expression of occludin protein， and the relative expressions of ZO-1 and Bcl-2 protein （P < 0.05）. Moreover， it presented an increase in permeability， apoptosis rate， MDA content， IL-6 and IL-1β levels， the relative expression of Bax protein， and the ratios of p-IKKβ/IKKβ， p-IκB-α/IκB-α， and p-NF-κB p65/NF-κB p65 （P < 0.05）. Conclusion Crocin alleviates the damage to glomerular endothelial cells induced by high glucose by inhibiting the activation of the IKKβ/NF-κB signaling pathway.