Statins inhibit protein kinase D (PKD) activation in intestinal cells and prevent PKD1-induced growth of murine enteroids.
Statins inhibit protein kinase D (PKD) activation in intestinal cells and prevent PKD1-induced growth of murine enteroids.
复制标题
他汀类药物可抑制肠细胞中蛋白激酶 D (PKD) 的激活，并阻止 PKD1 诱导的小鼠肠样细胞的生长。
DOI：
10.1152/ajpcell.00286.2022
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Rozengurt,Enrique
中科院分区：
文献类型：
--
作者：
Sinnett-Smith,James;Torres-Marquez,MEugenia;Chang,Jen-Kuan;Shimizu,Yuki;Hao,Fang;Martin,MartinG;Rozengurt,Enrique
我们检测了他汀类药物对G蛋白偶联受体激动剂激活蛋白激酶D(PKD)的影响。西立伐他汀可剂量依赖性地抑制血管紧张素Ⅱ(Ang II)或加压素诱导的肠上皮细胞系IEC-18细胞Ser916蛋白的蛋白脱氢酶(PKD)自磷酸化，在0.2µM时抑制作用减半最大。其他亲脂性的他汀类药物，包括辛伐他汀、阿托伐他汀和氟伐他汀，也以剂量依赖的方式阻止PKD的激活。使用标记了增强型绿色荧光蛋白(EGFP)的表达PKD1的IEC-18细胞株，西立伐他汀或辛伐他汀阻断了GPCR介导的PKD1-EGFP转位到质膜及其随后的核聚集。在表达PKD3-EGFP的IEC-18细胞中也得到了类似的结果。在机制上，他汀类药物抑制依赖激动剂的PKD激活，而不是直接作用于PKD的催化活性，因为暴露于西伐他汀或辛伐他汀不会损害PKD的自磷酸化或佛波二丁酸酯反应的PKD1-EGFP膜转位，后者绕过GPCRs，直接刺激PKC和PKD。此外，西立伐他汀不抑制通过体外激酶试验确定的重组PKD活性。使用来自PKD1转基因小鼠的肠隐窝上皮细胞生成的肠样细胞作为肠道再生的模型，我们发现他汀类药物对抗PKD1介导的肠样面积、复杂性(隐窝样芽的数量)和DNA合成的增加。我们的结果显示，他汀类药物对受体介导的PKD激活以及在对抗PKD1对肠上皮细胞的生长促进作用方面具有先前未被认识的抑制作用。
We examined the impact of statins on protein kinase D (PKD) activation by G protein-coupled receptor (GPCR) agonists. Treatment of intestinal IEC-18 cells with cerivastatin inhibited PKD autophosphorylation at Ser916induced by angiotensin II (ANG II) or vasopressin in a dose-dependent manner with half-maximal inhibition at 0.2 µM. Cerivastatin treatment inhibited PKD activation stimulated by these agonists for different times (5–60 min) and blunted HDAC5 phosphorylation, a substrate of PKD. Other lipophilic statins, including simvastatin, atorvastatin, and fluvastatin also prevented PKD activation in a dose-dependent manner. Using IEC-18 cell lines expressing PKD1 tagged with EGFP (enhanced green fluorescent protein), cerivastatin or simvastatin blocked GPCR-mediated PKD1-EGFP translocation to the plasma membrane and its subsequent nuclear accumulation. Similar results were obtained in IEC-18 cells expressing PKD3-EGFP. Mechanistically, statins inhibited agonist-dependent PKD activation rather than acting directly on PKD catalytic activity since exposure to cerivastatin or simvastatin did not impair PKD autophosphorylation or PKD1-EGFP membrane translocation in response to phorbol dibutyrate, which bypasses GPCRs and directly stimulates PKC and PKD. Furthermore, cerivastatin did not inhibit recombinant PKD activity determined via an in vitro kinase assay. Using enteroids generated from intestinal crypt-derived epithelial cells from PKD1 transgenic mice as a model of intestinal regeneration, we show that statins oppose PKD1-mediated increase in enteroid area, complexity (number of crypt-like buds), and DNA synthesis. Our results revealed a previously unappreciated inhibitory effect of statins on receptor-mediated PKD activation and in opposing the growth-promoting effects of PKD1 on intestinal epithelial cells.