Synergistic effect of schizandrin A and DNase I knockdown on high glucose induced beta cell apoptosis by decreasing intracellular calcium concentration.
Zhu Bin, Y U Ning, Wang Lei, Tian Yue, W U Mingfen, Zhao Zhigang
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引用次数: 0
Abstract
Objective: To explore the synergistic effect of deoxyribonuclease I (DNase I) knockdown combined with Schizandrin A (Sch A) in protecting islet beta-cells (β-cells) from apoptosis under high-glucose (HG) conditions.
Methods: The concentration of Sch A was detected by Cell Counting Kit-8 (CCK-8). High glucose-cultured rat insulinoma beta cell line (RIN-M5F) cells were treated with Sch A and transfected with DNase I small interfering RNA (siRNA). Cell apoptosis rate and apoptosis-related protein level were examined by flow cytometry and Western blot method respectively. In addition, Na-K-adenosine triphosphatease (Na-K-ATPase) and Ca-Mg-ATPase activity, cell membrane potential, and intracellular Ca concentration was also examined respectively.
Results: Our study revealed that HG stimulation can cause a significant increase in DNase I level and cell apoptosis rate. However, Sch A combined with DNase I knockdown can significantly decrease the cell apoptosis rate and apoptosis-related protein levels such as BAX ( 0.05) and Caspase-3 ( 0.01). In addition, we also found that the combination of Sch A and DNase I knockdown can dramatically increase cell membrane potential level, Na-K-ATPase, and Ca-Mg-ATPase activity. Meanwhile, intracellular Ca concentration was also found to be significantly decreased by the synergistic effect of Sch A and DNase I knockdown.
Conclusion: Overall, our study reveals a synergistic effect of Sch A and DNase I knockdown in protecting β-cells from HG-induced apoptosis.
目的探讨在高葡萄糖（HG）条件下，脱氧核糖核酸酶 I（DNase I）敲除与五味子甲（Sch A）联合使用对保护胰岛β细胞（β细胞）免于凋亡的协同作用：方法：用细胞计数试剂盒-8（CCK-8）检测 Sch A 的浓度。用 Sch A 处理高糖培养的大鼠胰岛素瘤 beta 细胞系（RIN-M5F）细胞，并用 DNase I 小干扰 RNA（siRNA）转染。分别用流式细胞术和 Western 印迹法检测细胞凋亡率和凋亡相关蛋白水平。此外，还分别检测了Na-K-腺苷三磷酸酶（Na-K-ATPase）和Ca-Mg-ATPase活性、细胞膜电位和细胞内Ca浓度：结果：我们的研究发现，HG 刺激可导致 DNase I 水平和细胞凋亡率显著增加。结果：我们的研究发现，HG 刺激可导致 DNase I 水平和细胞凋亡率显著升高，但 Sch A 与 DNase I 基因敲除结合使用可显著降低细胞凋亡率和凋亡相关蛋白水平，如 BAX（0.05）和 Caspase-3 （0.01）。此外，我们还发现 Sch A 和 DNase I 的联合敲除可大幅提高细胞膜电位水平、Na-K-ATPase 和 Ca-Mg-ATPase 活性。同时，在 Sch A 和 DNase I 基因敲除的协同作用下，细胞内 Ca 浓度也显著降低：总之，我们的研究揭示了 Sch A 和 DNase I 基因敲除在保护β细胞免受 HG 诱导的细胞凋亡方面的协同作用。
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