Histone-deacetylase-inhibitory effects of periodontopathic-bacterial metabolites induce human gingival epithelial Ca9-22 cell death
Histone-deacetylase-inhibitory effects of periodontopathic-bacterial metabolites induce human gingival epithelial Ca9-22 cell death
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DOI：
10.1007/s10266-022-00775-9
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发表时间：
2022-12
期刊：
影响因子：
2.5
通讯作者：
Kazuki Uemichi;Yoshikazu Mikami;Takayasu Watanabe;K. Shinozuka;M. Tonogi;Hiromasa Tsuda
中科院分区：
文献类型：
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作者：
Kazuki Uemichi;Yoshikazu Mikami;Takayasu Watanabe;K. Shinozuka;M. Tonogi;Hiromasa Tsuda
牙菌斑细菌产生高浓度的短链脂肪酸（SCFAs），作为细菌代谢产物。scfa处理的牙龈上皮细胞发生细胞死亡。我们之前的报告表明，丁酸盐诱导的细胞死亡依赖于自噬和活性氧（ROS）。然而，scfa诱导的牙龈上皮细胞死亡的确切机制尚不清楚。丁酸盐是一种强的组蛋白去乙酰化酶（HDAC）抑制剂。因此，我们确定了HDAC抑制活性在scfa诱导的牙龈上皮细胞中的作用。Ca9-22细胞被用作牙龈上皮细胞的体外对应物。在P300组蛋白乙酰转移酶（HAT）抑制剂C646存在或不存在的情况下，用HDAC抑制剂处理Ca9-22细胞，并比较使用SYTOX Green染料测量的死亡细胞数量。western blotting检测组蛋白H3乙酰化水平。利用RNA测序分析检测丁酸盐和C646处理期间转录组的变化。Ca9-22细胞的丁酸盐或丙酸处理诱导组蛋白H3的乙酰化，而C646处理强烈降低升高的乙酰化水平。因此，C646处理能抑制丁酸盐或丙酸盐诱导的细胞死亡。当使用其他HDAC抑制剂时，获得了类似的结果。全转录组分析显示，丁酸盐诱导的组蛋白乙酰化改变了许多基因的表达。此外，在改变基因中发现的一些自噬和ros相关基因可能诱导细胞死亡。本研究提示，细菌代谢产物需要hdac抑制活性来诱导细胞死亡，其作用可能会增强自噬和ROS的产生。
Dental plaque bacteria produce high concentrations of short-chain fatty acids (SCFAs), as bacterial metabolites. SCFA-treated gingival epithelial cells undergo cell death. Our previous reports demonstrated that butyrate-induced cell death depends on autophagy and reactive oxygen species (ROS). However, the precise mechanisms underlying SCFA-induced gingival epithelial cell death is poorly understood. Butyrate is a strong histone deacetylase (HDAC) inhibitor. Therefore, we determined the involvement of HDAC inhibitory activity in SCFA-induced gingival epithelial cells. Ca9-22 cells were used as an in vitro counterpart of gingival epithelial cells. Ca9-22 cells were treated with HDAC inhibitors in the presence or absence of C646, a P300 histone acetyltransferase (HAT) inhibitor, and compared the number of dead cells, which are measured using SYTOX Green dye. Acetylation levels of histone H3 were examined using western blotting. Changes in transcriptomes during the butyrate and C646 treatment were examined using RNA sequencing analysis. The butyrate or propionate-treatment of Ca9-22 cells induced acetylation of histone H3, while the C646 treatment strongly reduced the elevated acetylation levels. Accordingly, butyrate or propionate-induced cell death was inhibited by the C646 treatment. Similar results were obtained when other HDAC inhibitors were used. Whole transcriptome analysis revealed that the expression of numerous genes was altered by butyrate-induced histone acetylation. Moreover, some autophagy and ROS-related genes found in the altered genes might induce cell death. This study suggests the need for HDAC-inhibitory activity of bacterial metabolites to induce cell death, and the effects might enhance autophagy and ROS production.