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目的 探讨纺锤体和着丝粒相关蛋白复合物3(SKA3)调控低氧诱导的心肌凋亡的作用机制。方法 采用CoCl2·6H2O处理的人AC16心肌细胞作为细胞低氧的体外模型，体内实验使用C57BL/6J小鼠低氧环境4周构建体内低氧模型，小鼠共6只，均分为常氧组、低氧组。采用CCK-8评估细胞活力，通过比色法检测乳酸脱氢酶(LDH)以及天冬氨酸氨基转移酶(AST)水平以反应心肌细胞损伤。采用苏木精-伊红染色(HE)和免疫组化(IHC)观察小鼠心肺形态学变化，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)检测细胞凋亡，通过RT-qPCR和Westernblot检测缺氧诱导因子-1α(HIF-1α)、SKA3、凋亡相关因子如B细胞淋巴瘤-2(Bcl-2、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase3)mRNA和蛋白相对表达量。通过siRNA干扰SKA3表达，探究其在低氧诱导心肌细胞凋亡中的作用机制。结果 600μmol/LCoCl2·6H2O处理24h后细胞活力显著降低，HIF-1α表达水平升高。低氧组AC16细胞及小鼠心脏组织中AST与LDH表达水平显著升高，HE及IHC呈现典型的低氧性心肺损伤形态学改变。低氧模型中，SKA3表达显著升高，凋亡相关因子Bax、Caspase3表达升高、Bcl-2表达降低；siRNA干扰SKA3可显著逆转上述凋亡相关指标变化(均P<0.05)。结论 SKA3可通过低氧诱导因子HIF-1α调控低氧诱导的心肌细胞凋亡。
Abstract：Objective To explore the mechanism by which spindle- and kinetochore-associated complex subunit 3 (SKA3) regulates hypoxia-induced cardiomyocyte apoptosis. Methods Human AC16 cardiomyocytes treated with CoCl₂·6H₂O were used as an in-vitro cellular hypoxia model. For in-vivo experiments， C57BL/6J mice were exposed to hypoxic conditions for 4 weeks to establish an in-vivo hypoxia model. A total of 6 mice were equally divided into normoxia group (n = 3) and hypoxia group (n = 3). Cell viability was assessed by the CCK-8 assay. Lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) levels were measured by colorimetric assay to reflect cardiomyocyte injury. Hematoxylin-eosin (HE) staining and immunohistochemistry (IHC) were performed to observe morphological changes in mouse heart and lung tissues. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was applied to detect cell apoptosis. The relative mRNA and protein expression levels of hypoxia-inducible factor-1α (HIF-1α)， SKA3， and apoptosis-related factors including B-cell lymphoma-2 (Bcl-2)， Bcl-2-associated X protein (Bax)， and cysteinyl aspartate-specific protease-3 (Caspase-3) were determined by RT-qPCR and Western blot. siRNA-mediated SKA3 knockdown was used to investigate its functional mechanism in hypoxia-triggered cardiomyocyte apoptosis. Results Treatment with 600 μmol/L CoCl₂·6H₂O for 24 h markedly reduced cell viability and up-regulated HIF-1α expression. AST and LDH levels were significantly elevated in hypoxic AC16 cells and cardiac tissues from hypoxic mice. HE staining and IHC revealed typical morphological features of hypoxic heart-lung injury. In hypoxic models， SKA3 expression was notably increased， accompanied by up-regulation of Bax and Caspase-3 as well as down-regulation of Bcl-2. siRNA-mediated SKA3 silencing significantly reversed these alterations in apoptosis-related markers (all P < 0.05). Conclusion SKA3 modulates hypoxia-induced cardiomyocyte apoptosis via hypoxia-inducible factor signaling.
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基本信息:
中图分类号:R594.3
引用信息:
[1]毛亚琪,王士伟,靳子冲,等.SKA3在高原低氧致心肌凋亡的作用机制研究[J].安徽医科大学学报().
基金信息:
国家自然科学基金项目（编号：82460333、U20A20395）; 西藏自治区科技计划项目（编号：XZ202401JD0031）
2026-09-11
2026-09-11
2026-09-11