Inhibition of Calcineurin and Glycogen Synthase Kinase‐3β by Ricinoleic Acid Derived from Castor Oil
Inhibition of Calcineurin and Glycogen Synthase Kinase‐3β by Ricinoleic Acid Derived from Castor Oil
复制标题
蓖麻油中的蓖麻油酸对钙调神经磷酸酶和糖原合酶激酶 3β 的抑制作用
DOI：
10.1002/lipd.12208
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发表时间：
2020
期刊：
影响因子：
1.9
通讯作者：
Kimura Ken‐ichi
中科院分区：
文献类型：
--
作者：
Yoshida Jun;Okawa Yusuke;Oyama Takuya;Shimoda Nozomu;Uesugi Shota;Takagi Hiroshi;Ito Yoshiaki;Kimura Ken‐ichi
蓖麻油酸（RA）是蓖麻油的主要脂肪酸成分，在基于酵母的药物筛选试验中发现其可抑制Ca 2+信号转导途径介导的细胞周期调节。预计RA具有抗糖尿病、抗过敏和/或抗癌特性，但其靶分子未知。为了确定RA的新药理作用，我们研究了其在Ca 2+信号转导途径中的靶分子。在使用thersp 5A 401 E突变体的基于酵母的CN抑制剂筛选试验和使用重组人CN的磷酸酶试验中检查了RA对钙调磷酸酶（CN）的抑制。在使用thersp 5A 401 E酵母菌株的CN抑制剂筛选试验中，RA在5 μg/斑点下显示出生长恢复活性。此外，在Ki = 33.7 μM时，它以底物竞争性方式直接抑制CN而不含亲免素。通过测量RBL-2 H3细胞中β-己糖胺酶（β-HEX）的释放，进一步评价了RA对哺乳动物细胞中CN的影响。50 μM RA抑制RBL-2 H3细胞释放β-HEX。此外，通过使用重组人GSK-3β进行的激酶测定，发现该化合物可抑制糖原合成酶激酶-3 β（GSK-3β）。RA在Ki = 1.43 μM时以肽底物竞争性方式抑制GSK-3β。通过测量H4 IIE大鼠肝癌细胞中葡萄糖产生的抑制作用，在哺乳动物细胞中进一步评估了该分子对GSK-3β的抑制作用。25 μM RA抑制这些细胞中葡萄糖的产生。这些发现表明RA和/或蓖麻油可能是治疗过敏或2型糖尿病的有用功能性脂肪酸。
Ricinoleic acid (RA) is the main fatty acid component of castor oil and was found to inhibit Ca2+‐signal transduction pathway‐mediated cell cycle regulation in a yeast‐based drug screening assay. RA is expected to have antidiabetic, antiallergy, and/or anticancer properties but its target molecule is unknown. To identify a novel pharmacological effect of RA, we investigated its target molecule in the Ca2+‐signal transduction pathway. RA inhibition of calcineurin (CN) was examined in a yeast‐based CN inhibitor screening assay using thersp5A401Emutant and in a phosphatase assay using recombinant human CN. RA showed growth‐restoration activity at 5 μg/spot in the CN inhibitor screening assay with thersp5A401Eyeast strain. Furthermore, it directly inhibited CN without immunophilins atKi= 33.7 μM in a substrate‐competitive manner. The effects of RA on CN in mammalian cells were further evaluated by measuring β‐hexosaminidase (β‐HEX) release in RBL‐2H3 cells. RA at 50 μM suppressed the release of β‐HEX from RBL‐2H3 cells. Moreover, this compound was found to inhibit glycogen synthase kinase‐3β (GSK‐3β), as determined by a kinase assay using recombinant human GSK‐3β. RA inhibited GSK‐3β atKi= 1.43 μM in a peptide substrate‐competitive manner. The inhibition of GSK‐3β by this molecule was further assessed in mammalian cells by measuring the inhibition of glucose production in H4IIE rat hepatoma cells. RA at 25 μM suppressed glucose production in these cells. These findings indicate that RA and/or castor oil could be a useful functional fatty acid to treat allergy or type 2 diabetes.