Funamoto Masafumi, Sunagawa Yoichi, Gempei Mai, Shimizu Kana, Katanasaka Yasufumi, Shimizu Satoshi, Hamabe-Horiike Toshihide, Appendino Giovanni, Minassi Alberto, Koeberle Andreas, Komiyama Maki, Mori Kiyoshi, Hasegawa Koji, Morimoto Tatsuya
Division of Molecular Medicine, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
Division of Translational Research, Clinical Research Institute, National Hospital Organization Kyoto Medical Center, 1-1 Fukakusa Mukaihatacho, Fushimi-ku, Kyoto 612-8555, Japan.
Pharmaceutics. 2022 Jun 15;14(6):1269. doi: 10.3390/pharmaceutics14061269.
The intrinsic histone acetyltransferase (HAT), p300, has an important role in the development and progression of heart failure. Curcumin (CUR), a natural p300-specific HAT inhibitor, suppresses hypertrophic responses and prevents deterioration of left-ventricular systolic function in heart-failure models. However, few structure-activity relationship studies on cardiomyocyte hypertrophy using CUR have been conducted. To evaluate if prenylated pyrazolo curcumin (PPC) and curcumin pyrazole (PyrC) can suppress cardiomyocyte hypertrophy, cultured cardiomyocytes were treated with CUR, PPC, or PyrC and then stimulated with phenylephrine (PE). PE-induced cardiomyocyte hypertrophy was inhibited by PyrC but not PPC at a lower concentration than CUR. Western blotting showed that PyrC suppressed PE-induced histone acetylation. However, an in vitro HAT assay showed that PyrC did not directly inhibit p300-HAT activity. As Cdk9 phosphorylates both RNA polymerase II and p300 and increases p300-HAT activity, the effects of CUR and PyrC on the kinase activity of Cdk9 were examined. Phosphorylation of p300 by Cdk9 was suppressed by PyrC. Immunoprecipitation-WB showed that PyrC inhibits Cdk9 binding to CyclinT1 in cultured cardiomyocytes. PyrC may prevent cardiomyocyte hypertrophic responses by indirectly suppressing both p300-HAT activity and RNA polymerase II transcription elongation activity via inhibition of Cdk9 kinase activity.
内在组蛋白乙酰转移酶(HAT)p300在心力衰竭的发生和发展中起重要作用。姜黄素(CUR)是一种天然的p300特异性HAT抑制剂,可抑制肥厚反应并预防心力衰竭模型中左心室收缩功能的恶化。然而,很少有使用CUR对心肌细胞肥大进行构效关系研究。为了评估异戊烯基吡唑姜黄素(PPC)和姜黄素吡唑(PyrC)是否能抑制心肌细胞肥大,用CUR、PPC或PyrC处理培养的心肌细胞,然后用去甲肾上腺素(PE)刺激。在比CUR更低的浓度下,PyrC可抑制PE诱导的心肌细胞肥大,而PPC则不能。蛋白质印迹法显示,PyrC可抑制PE诱导的组蛋白乙酰化。然而,体外HAT分析表明,PyrC不会直接抑制p300-HAT活性。由于Cdk9可使RNA聚合酶II和p300磷酸化并增加p300-HAT活性,因此研究了CUR和PyrC对Cdk9激酶活性的影响。PyrC可抑制Cdk9对p300的磷酸化。免疫沉淀-蛋白质印迹法显示,PyrC可抑制培养心肌细胞中Cdk9与细胞周期蛋白T1的结合。PyrC可能通过抑制Cdk9激酶活性间接抑制p300-HAT活性和RNA聚合酶II转录延伸活性,从而预防心肌细胞肥大反应。