河北医科大学学报 ›› 2025, Vol. 46 ›› Issue (1): 5-13.doi: 10.3969/j.issn.1007-3205.2025.01.002

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蛋白激酶C抑制剂对苏尼替尼所致心脏毒性的保护作用

  

  1. 1.河北医科大学药理教研室，河北省新药药理毒理重点实验室，教育部血管与神经生物重点实验室，
    河北 石家庄 050017；2.河北医科大学第四医院心内科，河北 石家庄 050011

  • 出版日期:2025-01-25 发布日期:2025-01-22
  • 作者简介:邱素华(1984-)，女，河南鹿邑人，河北医科大学实验师，理学硕士，从事心血管药理毒理研究。
  • 基金资助:
    河北省自然科学基金精准医学联合基金重点项目（H2020206554）

Protective effect of protein kinase C inhibitors on sunitinib-induced cardiotoxicity

  1. 1.Department of Pharmacology, Hebei Medical University， the Key Laboratory of New Drug 
    Pharmacology and Toxicology of Hebei Province， the Key Laboratory of Neural and 
    Vascular Biology, Ministry of Education, Shijiazhuang 050017, China; 
    2.Department of Cardiology, the Fourth Hospital of Hebei Medical 
    University, Shijiazhuang 050011, China

  • Online:2025-01-25 Published:2025-01-22

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摘要： 目的 在细胞和整体动物水平探讨蛋白激酶C（protein kinase C，PKC）抑制剂对苏尼替尼（Sunitinib, SU）引发心脏毒性的保护作用。
方法 新生大鼠心肌细胞（neonatal rat ventricular myocytes，NRVMs）在不同浓度的SU(1,5,10 μmol/L)中进行孵育, 检测细胞三磷酸腺苷（adenosine triphosphate，ATP）含量、乳酸脱氢酶（lactate dehydrogenase，LDH）释放量及线粒体膜电位(mitochondrial membrane potential，MMP)。然后观察非选择性PKC抑制剂（bisindolylmaleimide 1，Bis-1）、选择性新型PKC抑制剂（Rotterlin）或抑制肽对SU引发心肌细胞毒性的影响。雄性C57小鼠随机分为4组，即正常对照组、SU组、SU+PKC抑制剂白屈菜红碱（chelerythrine,CHE）低剂量（0.375 mg·kg-1·d-1）及CHE高剂量（0.75 mg·kg-1·d-1）组。连续灌胃SU同时腹腔注射CHE，给药3周后取心脏组织，提取线粒体，检测心肌细胞线粒体呼吸链复合物Ⅳ和Ⅴ活性；电镜观察心肌细胞线粒体结构变化；Western blot检测各组心肌磷酸化PKC亚型包括PKCα、PKCε以及PKCδ的表达情况。
结果 SU孵育NRVMs 后呈浓度与时间依赖性减少ATP含量、增加LDH释放量、降低线粒体膜电位。非选择性PKC抑制剂 Bis-1可预防SU 引发的上述改变；选择性新型PKC抑制剂Rotterlin和新型PKCε抑制肽亦可预防SU引起的上述各项指标的变化。小鼠给予SU后，心肌细胞线粒体呼吸链复合物Ⅳ和Ⅴ的活性均显著下降，电镜显示，心肌细胞线粒体出现明显肿胀、嵴融合等形态改变；同时给予不同剂量的CHE后，线粒体呼吸链复合物Ⅳ和Ⅴ的活性均显著升高，电镜结果显示，CHE明显减轻SU所致线粒体形态改变。Western 检测显示， SU组心肌磷酸化PKCε表达量显著升高，而磷酸化PKCδ、PKCα表达量则无明显变化；高剂量的CHE显著抑制了磷酸化PKCε的过表达，使其恢复至对照组水平。
结论 PKC抑制剂对SU导致的心脏毒性具有显著保护作用，此作用可能源于抑制新型PKC（主要是PKCε亚型）的过度激活。


关键词: 蛋白激酶C抑制剂, 苏尼替尼, 心脏毒性

Abstract: Objective To investigate the protective effects of protein kinase C(PKC)inhibitors on the cardiotoxicity induced by sunitinib (SU) at the cellular and whole animal levels. 
Methods The neonatal rat ventricular myocytes (NRVMs) were cultured and incubated with different concentrations of SU (1, 5, 10 μmol/L). The intracellular adenosine triphosphate (ATP) level, lactate dehydrogenase (LDH) release and the mitochondrial membrane potential (MMP) were measured. Then the effects of non-selective PKC inhibitor bisindolylmaleimide 1 (Bis-1), selective novel PKC inhibitor Rotterlin, or inhibitory peptides on SU-induced cardiotoxicity were observed. Male C57 mice were randomly divided into four groups, including control group, SU group, SU combined with PKC inhibitor chelerythrine (CHE) at low dose (i.p. 0.375 mg·kg-1·d-1) and high dose (i.p. 0.75 mg·kg-1·d-1) groups. SU was administered continuously by gavage and intraperitoneal injection of CHE was given. At 3 weeks after administration, the heart tissues were harvested and mitochondria from ventricular myocardium were extracted to measure the activity of mitochondrial complexes Ⅳ and Ⅴ. The transmission electron microscopy (TEM) was used to observe the mitochondria microstructures. Western blot analysis was performed to detect the expression of phosphorylated PKC subtypes, including p-PKCε, p-PKCδ and p-PKCα in each group. 
Results SU significantly reduced ATP level, increased LDH release and decreased MMP in NRVMs in a concentration and time-dependent manner. TEM showed that SU-induced visible changes could be prevented by non-selective PKC inhibitor Bis-1, selective novel PKC inhibitor Rotterlin and selective PKCε inhibitory peptide. Administration of SU significantly reduced the activity of mitochondrial complexes Ⅳ and Ⅴ in mouse myocardium, and TEM showed significant morphological changes such as swelling and cristae fusion in myocardial cell mitochondria. Co-administration of different dose of CHE prevented the decrease of mitochondrial complexes Ⅳ and Ⅴ activities. Meanwhile, TEM showed that CHE significantly alleviated the mitochondrial morphological changes induced by SU. Western blot analysis showed that the expression level of p-PKCε was significantly increased in SU group compared with the control group, but p-PKCδ or p-PKCα had no significant change. High-dose CHE significantly inhibited the overexpression of p-PKCε, restoring it to the control group level. 
Conclusion PKC inhibitors have a significant protective effect against SU-induced cardiotoxicity, which may be due to the inhibition of excessive activation of novel PKC (mainly PKCε subtype). 


Key words: protein kinase C inhibitor, sunitinib, cardiotoxicity

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