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目的：探讨非肌肉肌球蛋白ⅡA(non-muscle myosinⅡA, NMⅡA)在高糖（high glucose,HG）诱导的脑微血管内皮细胞损伤中的作用机制。方法：采用高糖培养基培养小鼠脑微血管内皮细胞bEnd.3，建立体外糖尿病脑血管病变模型。利用siRNA技术敲低NMⅡA表达，检测细胞活力、炎症因子、氧化应激产物水平及细胞表征改变。通过CCK-8法、细胞划痕实验、原子力显微镜检测等方法，检测细胞增殖、迁移、侵袭及力学特性变化。通过Western blot和RT-qPCR检测相关蛋白及基因表达。结果：成功构建bEnd.3高糖模型。敲低NMⅡA后，炎症因子和氧化应激产物水平明显升高，细胞活力和迁移能力降低，细胞高度、表面黏附力和杨氏模量降低（P<0.05）。结论：NMⅡA参与细胞收缩、迁移、胞质分裂及维持细胞张力等关键过程。在高糖环境下，敲低NMⅡA会造成炎症因子等指标加重，细胞功能活力减弱（P<0.05）。说明NMⅡA或是维持高糖状态下脑微血管内皮细胞正常功能的关键因素之一，可能为治疗糖尿病脑血管病变寻找到新的潜在靶点。
Abstract：Objective: To investigate the mechanism of non-muscle myosin ⅡA(NMⅡA) in high glucose(HG)-induced injury of brain microvascular endothelial cells bEnd.3. Methods: An in vitro diabetic cerebrovascular injury model was established by culturing bEnd.3 cells in HG medium. NMⅡA expression was knocked down using siRNA, followed by assessments of cell viability, inflammatory factors, oxidative stress markers, and cellular morphological changes. Functional assays(CCK-8, wound healing, atomic force microscopy) were performed to evaluate proliferation, migration, invasion, and mechanical properties. Protein and gene expression were analyzed by Western blot and RT-qPCR. Results: The HG-induced bEnd.3 model was successfully established. NMⅡA knockdown exacerbated inflammation and oxidative stress, reduced cell viability and migration, and decreased cell height, adhesion, and Young's modulus(P<0.05). Conclusion: NM Ⅱ A regulates critical processes including cell contraction, migration, and tension maintenance. Under HG conditions, NMⅡA deficiency worsens inflammation and impairs endothelial function, suggesting its pivotal role in preserving cerebrovascular homeostasis(P<0.05). Targeting NMⅡA may offer a novel therapeutic strategy for diabetic cerebrovascular complications.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20250926.001
中图分类号:R587.2;R743
引用信息:
[1]赵玉洁,谭越雯,薛晋垣,等.非肌肉肌球蛋白ⅡA维持高糖诱导的脑微血管内皮细胞功能[J].海南医科大学学报,2026,32(08):561-569.DOI:10.13210/j.cnki.jhmu.20250926.001.
基金信息:
国家自然科学基金（32000551）~~
2025-09-26
2025-09-26
2025-09-26