Wen Shi-Fang, Tang Zhi-Yuan, Shen Xian-Juan, Chen Tao, Zhao Jian-Mei
Department of Pediatrics, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2025 Jul 15;27(7):842-849. doi: 10.7499/j.issn.1008-8830.2412085.
To investigate the role and mechanism of copper overload-mediated endoplasmic reticulum stress (ERS) in vascular endothelial injury in Kawasaki disease (KD).
Four-week-old male C57BL/6 mice were randomly divided into four groups: control, KD, KD plus copper chelator tetrathiomolybdate (TTM), and KD plus ERS inhibitor AMG PERK 44 (AMG) (=20 per group). A KD mouse model was established using Candida albicans extract. Human umbilical vein endothelial cells (HUVECs) were divided into control (intervention with healthy children's serum), KD (intervention with KD patients' serum), and KD+TTM (intervention with KD patients' serum plus 20 µmol/L TTM). Copper deposition in mouse heart tissue was assessed using rubeanic acid staining. Vascular pathological changes were observed using hematoxylin-eosin staining and measurement of abdominal aortic diameter and area. ERS activation was detected by transmission electron microscopy and immunofluorescence. HUVEC viability, apoptosis, and functional changes were evaluated using CCK8, flow cytometry, cell scratch assay, and angiogenesis experiments. ERS marker protein expression levels were measured by Western blot.
Compared to the KD group, the KD+TTM and KD+AMG groups showed reduced copper deposition in the vascular wall, decreased swelling of coronary endothelial cells and endoplasmic reticulum, reduced inflammatory cell infiltration, and less abdominal aortic lesion expansion. The abdominal aortic diameter and area, and the fluorescence intensity of ERS marker proteins (GRP78 and CHOP) were significantly lower (<0.05). Compared to the KD group, the KD+TTM group exhibited increased cell viability, tube number, and scratch healing rate, along with decreased apoptosis rate and expression of ERS marker proteins (GRP78, CHOP, ATF6, and p-PERK) (<0.05).
Copper overload aggravates vascular endothelial injury in KD by activating the ERS pathway. TTM can exert protective effects on the endothelium by regulating copper metabolism and inhibiting the ERS pathway.
探讨铜过载介导的内质网应激（ERS）在川崎病（KD）血管内皮损伤中的作用及机制。
将4周龄雄性C57BL/6小鼠随机分为四组：对照组、KD组、KD加铜螯合剂四硫代钼酸盐（TTM）组和KD加ERS抑制剂AMG PERK 44（AMG）组（每组n = 20）。用白色念珠菌提取物建立KD小鼠模型。人脐静脉内皮细胞（HUVECs）分为对照组（用健康儿童血清干预）、KD组（用KD患者血清干预）和KD + TTM组（用KD患者血清加20 μmol/L TTM干预）。用赤藓红酸染色评估小鼠心脏组织中的铜沉积。用苏木精-伊红染色观察血管病理变化，并测量腹主动脉直径和面积。通过透射电子显微镜和免疫荧光检测ERS激活情况。用CCK8、流式细胞术、细胞划痕试验和血管生成实验评估HUVEC活力、凋亡和功能变化。通过蛋白质免疫印迹法检测ERS标记蛋白表达水平。
与KD组相比，KD + TTM组和KD + AMG组血管壁铜沉积减少，冠状动脉内皮细胞和内质网肿胀减轻，炎症细胞浸润减少，腹主动脉病变扩张减轻。腹主动脉直径和面积以及ERS标记蛋白（GRP78和CHOP）的荧光强度显著降低（P < 0.05）。与KD组相比，KD + TTM组细胞活力、管腔数量和划痕愈合率增加，凋亡率和ERS标记蛋白（GRP78、CHOP、ATF6和p - PERK）表达降低（P < 0.05）。
铜过载通过激活ERS途径加重KD血管内皮损伤。TTM可通过调节铜代谢和抑制ERS途径对内皮发挥保护作用。
