Ministry of Education Laboratory of Combinatorial Biosynthesis and Drug Discovery, School of Pharmaceutical Science, Wuhan University, PR China.
Chem Biol Interact. 2011 Nov 15;194(2-3):127-33. doi: 10.1016/j.cbi.2011.09.005. Epub 2011 Oct 12.
Alpha-subtype protein kinase C (PKCα) is closely related to cardiovascular disease. Ritonavir (RTV), which is a human immunodeficiency virus (HIV) protease inhibitor, can induce atherosclerosis in a PKC-dependent manner. However, it remains unclear how RTV acts on PKCα to induce pathological phenotypes. In this study, we obtained mouse peritoneal macrophages from adult female Kunmin mice. The results of Oil Red O staining and immunofluorescence using confocal laser scanning microscope demonstrated that RTV could induce foam cell formation and plasma membrane translocation of PKCα like phorbol-12-myristate-13-acetate (PMA, a PKC activator). Computational modeling also exhibited similar docking of RTV and PMA to PKCα and similar patterns of hydrophobic interaction and hydrogen bond formation. Further in vitro kinase activity studies revealed that RTV could elevate PKC activity. These data provided insight into the PKC-dependent induction of atherosclerosis and useful information for more in-depth toxicity research of HIV protease inhibitor (PI). In addition, western blot analysis proved RTV also up-regulate PKCα expression, which may be related to its influence on estrogen responsiveness in target cells and needs further prove.
α 亚型蛋白激酶 C（PKCα）与心血管疾病密切相关。利托那韦（RTV）是一种人类免疫缺陷病毒（HIV）蛋白酶抑制剂，可通过依赖 PKC 的方式诱导动脉粥样硬化。然而，目前尚不清楚 RTV 如何作用于 PKCα 诱导病理表型。在本研究中，我们从小鼠腹腔巨噬细胞中获得了成年昆明雌性小鼠。油红 O 染色和共聚焦激光扫描显微镜的免疫荧光结果表明，RTV 可诱导泡沫细胞形成和 PKCα 的质膜易位，作用类似于佛波醇 12-肉豆蔻酸 13-乙酸酯（PKC 激活剂）。计算建模也显示了 RTV 和 PMA 与 PKCα 的类似对接以及疏水相互作用和氢键形成的类似模式。进一步的体外激酶活性研究表明，RTV 可提高 PKC 活性。这些数据为 PKC 依赖性动脉粥样硬化诱导提供了深入的见解，并为更深入的 HIV 蛋白酶抑制剂（PI）毒性研究提供了有用的信息。此外，Western blot 分析证明 RTV 还可上调 PKCα 的表达，这可能与其对靶细胞中雌激素反应性的影响有关，需要进一步证实。
