目的 探讨基于Kelch样环氧氯丙烷相关蛋白1（Keap1）/核因子E2相关因子2（Nrf2）/抗氧化反应元件（ARE）信号通路灯盏花素（Bre）对川崎病（KD）小鼠冠脉损伤的影响。 方法 除Control组外，构建KD小鼠并随机分为KD、L-Bre、M-Bre、H-Bre组（腹腔注射20、50、100 mg/kg的Bre）、Bre+ML385（腹腔注射100 mg/kg的Bre+30 mg/kg Nrf2抑制剂ML385）。HE染色观察冠状动脉组织病理变化；TUNEL染色观察冠状动脉细胞凋亡；ELISA检测血液心肌损伤因子[肌红蛋白（Mb）、肌酸激酶（CK）、肌酸激酶同工酶（CK-MB）]、氧化应激指标[活性氧（ROS）、超氧化物歧化酶（SOD）]、炎症因子[肿瘤坏死因子（TNF-α）、白细胞介素-1β（IL-1β）]的表达；Western blot检测冠脉组织Keap1、Nrf2、HO-1、NQO1蛋白表达。 结果 KD组Mb、CK、CK-MB、凋亡率、TNF-α、IL-1β、ROS、Keap1表达高于Control组，SOD、Nrf2、HO-1、NQO1表达低于Control组（P<0.05）；L-Bre、M-Bre、H-Bre组Mb、CK、CK-MB、凋亡率、TNF-α、IL-1β、ROS、Keap1低于KD组，SOD、Nrf2、HO-1、NQO1表达高于KD组（P<0.05）；Bre+ML385组上述指标变化趋势与H-Bre组相反（P<0.05）。 结论 Bre可能经Keap1/Nrf2/ARE信号通路抑制KD小鼠的冠脉损伤。
Abstract
Objective To investigate the effects of breviscapine (Bre) on coronary artery injury in Kawasaki disease (KD) mice based on the Kelch-like epichlorohydrin related protein 1 (Keap1)/nuclear factor erythroid 2 related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. Methods KD mice were constructed and randomly divided into the KD , the L-Bre, the M-Bre, the H-Bre groups (intraperitoneal injection of 20, 50, and 100 mg/kg of Bre), and the Bre + ML385 group (intraperitoneal injection of 100 mg/kg of Bre and 30 mg/kg Nrf2 inhibitor ML385). HE staining was applied to observe pathological changes in coronary artery tissue. TUNEL staining was used to observe apoptosis of artery tissue cells. ELISA was used to detect the expressions of cardiac injury factors in the blood [myoglobin (Mb), creatine kinase (CK), creatine kinase isoenzyme (CK-MB)], oxidative stress indicators [reactive oxygen species (ROS), superoxide dismutase (SOD)], and inflammatory factors [tumor necrosis factor (TNF-α), interleukin-1β (IL-1β)]. Western blot was applied to detect the expression of Keap1, Nrf2, HO-1, and NQO1 proteins in artery tissue. Results The Mb, CK, CK-MB, apoptosis rate, expression of TNF-α, IL-1β, ROS, and Keap1 in the KD group were higher than that in Control group, while the expression of SOD, Nrf2, HO-1 and NQO1 was lower than that in Control group (P<0.05). The Mb, CK, CK-MB, apoptosis rate, expression of TNF-α, IL-1β, ROS, and Keap1 in the L-Bre, M-Bre, and H-Bre groups were lower than that in KD group, while the expression of SOD, Nrf2, HO-1, and NQO1 was higher than that in KD group (P<0.05). The changes in the above indicators in the Bre+ML385 group were in the opposite direction compared to those in H-Bre group. Conclusions Bre can inhibit coronary artery injury in KD mice, which may be achieved by regulating the Keap1/Nrf2/ARE signaling pathway.
关键词
Kelch样环氧氯丙烷相关蛋白1 /
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核因子E2相关因子2 /
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抗氧化反应元件 /
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灯盏花素 /
川崎病 /
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冠脉损伤
Key words
Kelch-like epichlorohydrin related protein 1 /
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Nuclear factor erythroid 2 related factor 2 /
Antioxidant response element /
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Breviscapine /
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Kawasaki disease /
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Coronary artery injury
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基金
扬州市基础研究计划（联合专项）卫生健康类项目（2025-2-09）；江苏省妇幼保健科研项目（F202218）；扬州大学心脏大血管病研究院开放课题（YIHV0008）；扬州市医学重点学科（YZYXZDXK-15）
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