TSLP 通过人单核细胞中的组蛋白修饰调节线粒体 ROS 诱导的线粒体自噬
2022/04/19
摘要
背景：胸腺基质淋巴细胞生成素(Thymic stromal lymphoopoietin, TSLP)是一种Th2样细胞因子，参与哮喘的发病。过多的活性氧(ROS)产生可导致气道炎症、高反应性和气道重塑。ROS的产生会引起线粒体自噬。线粒体自噬是自噬对线粒体的选择性降解，通常发生在有缺陷的线粒体中。
目的：在本研究中，我们旨在研究TSLP对人单核细胞ROS产生和线粒体自噬的影响，并探讨包括表观遗传调控在内的其中潜在的机制。
结果：TSLP诱导THP-1细胞产生ROS，并被抗氧化剂N-乙酰半胱氨酸(NAC)逆转。透射电子显微镜图像显示出在TSLP刺激后失去嵴超微结构的环状线粒体。TSLP刺激后线粒体膜电位降低，MTCO2表达降低，线粒体DNA释放增加。TSLP增强了线粒体复合物I和复合物II/III的活性，增加了线粒体拷贝数和复合物II SHDA基因的表达。TSLP诱导的SHDA表达受到组蛋白乙酰转移酶抑制剂阿纳卡酸(AA)和组蛋白甲基转移酶抑制剂甲基硫代腺苷(MTA)的抑制。染色质免疫沉淀检测显示，TSLP增强了SHDA启动子的H3和H4的乙酰化、H3K4和H3K36的三甲基化。共聚焦激光显微镜显示，TSLP处理增加了线粒体自噬相关蛋白PINK1、LC3、磷酸帕金和磷酸泛素的信号，并且用AA和MTA预处理减少了TSLP诱导的PINK1和LC3在线粒体中的积累。Western印迹分析显示TSLP显著增加了磷-AMPK信号强度，而该作用可以被抗氧化剂NAC所抑制。AMPK抑制剂dorsomorphin降低了线粒体自噬相关蛋白PINK1、Parkin和LC3 I/II的信号强度。TSLP可以减少M1相关细胞因子CXCL-10的产生，并增加M2相关细胞因子CCL-1和CCL-22的产生，而这种效应可以被线粒体自噬抑制剂Mdivi-1和 PINK1基因敲低所抑制。
结论：上皮来源的TSLP通过AMPK激活和组蛋白修饰可以调节ROS的产生和线粒体自噬，并可以改变人单核细胞中M1/M2趋化因子的表达。
TSLP regulates mitochondrial ROS-induced mitophagy via histone modification in human monocytes
Lin YC, Lin YC, Tsai ML, et al.
Abstract
BACKGROUNDS：Thymic stromal lymphopoietin (TSLP) is a Th2-like cytokine involved in asthma pathogenesis. Excessive reactive oxygen species (ROS) production can lead to airway inflammation, hyperresponsiveness and remodeling. Mitophagy, followed by ROS production, is the selective degradation of mitochondria by autophagy and often occurs in defective mitochondria.
OBJECTIVE：In the present study, we aimed to examine the effects of TSLP on ROS production and mitophagy in human monocytes and to investigate the underlying mechanisms, including epigenetic regulation.
RESULTS：TSLP induced ROS generation, and the effects were reversed by the antioxidant N-acetylcysteine (NAC) in THP-1 cells. Transmission electron microscopy images showed donut-shaped mitochondria that lost the cristae ultrastructure after TSLP stimulation. A decrease in mitochondrial membrane potential, decreased MTCO2 expression, and increased mitochondrial DNA release after TSLP stimulation were found. TSLP enhanced mitochondrial complex I and complex II/III activity and increased mitochondrial copy numbers and the expression of the complex II SHDA gene. TSLP-induced SHDA expression was inhibited by the histone acetyltransferase inhibitor anacardic acid (AA) and the histone methyltransferase inhibitor methylthioadenosine (MTA), and chromatin immunoprecipitation assays revealed that TSLP enhanced H3 acetylation, H4 acetylation, and H3K4 and H3K36 trimethylation in the SHDA promoter. Confocal laser microscopy showed that TSLP treatment increased the signals of the mitophagy-related proteins PINK1, LC3, phospho-parkin and phospho-ubiquitin, and pretreatment with AA and MTA reduced TSLP-induced PINK1 and LC3 accumulation in mitochondria. Western blot analysis showed that TSLP significantly increased phosphor-AMPK signal intensity, and the effects were inhibited by the antioxidant NAC. The increased signal intensities of the mitophagy-related proteins PINK1, Parkin and LC3 I/II were decreased by dorsomorphin, an AMPK inhibitor. TSLP decreased M1-related cytokine CXCL-10 production and increased M2-related cytokine CCL-1 and CCL-22 production, which was suppressed by the mitophagy inhibitor Mdivi-1 and PINK1 gene knockdown.
CONCLUSIONS：Epithelial-derived TSLP regulates ROS production and mitophagy through AMPK activation and histone modification and alters M1/M2 chemokine expression in human monocytes.
背景：胸腺基质淋巴细胞生成素(Thymic stromal lymphoopoietin, TSLP)是一种Th2样细胞因子，参与哮喘的发病。过多的活性氧(ROS)产生可导致气道炎症、高反应性和气道重塑。ROS的产生会引起线粒体自噬。线粒体自噬是自噬对线粒体的选择性降解，通常发生在有缺陷的线粒体中。
目的：在本研究中，我们旨在研究TSLP对人单核细胞ROS产生和线粒体自噬的影响，并探讨包括表观遗传调控在内的其中潜在的机制。
结果：TSLP诱导THP-1细胞产生ROS，并被抗氧化剂N-乙酰半胱氨酸(NAC)逆转。透射电子显微镜图像显示出在TSLP刺激后失去嵴超微结构的环状线粒体。TSLP刺激后线粒体膜电位降低，MTCO2表达降低，线粒体DNA释放增加。TSLP增强了线粒体复合物I和复合物II/III的活性，增加了线粒体拷贝数和复合物II SHDA基因的表达。TSLP诱导的SHDA表达受到组蛋白乙酰转移酶抑制剂阿纳卡酸(AA)和组蛋白甲基转移酶抑制剂甲基硫代腺苷(MTA)的抑制。染色质免疫沉淀检测显示，TSLP增强了SHDA启动子的H3和H4的乙酰化、H3K4和H3K36的三甲基化。共聚焦激光显微镜显示，TSLP处理增加了线粒体自噬相关蛋白PINK1、LC3、磷酸帕金和磷酸泛素的信号，并且用AA和MTA预处理减少了TSLP诱导的PINK1和LC3在线粒体中的积累。Western印迹分析显示TSLP显著增加了磷-AMPK信号强度，而该作用可以被抗氧化剂NAC所抑制。AMPK抑制剂dorsomorphin降低了线粒体自噬相关蛋白PINK1、Parkin和LC3 I/II的信号强度。TSLP可以减少M1相关细胞因子CXCL-10的产生，并增加M2相关细胞因子CCL-1和CCL-22的产生，而这种效应可以被线粒体自噬抑制剂Mdivi-1和 PINK1基因敲低所抑制。
结论：上皮来源的TSLP通过AMPK激活和组蛋白修饰可以调节ROS的产生和线粒体自噬，并可以改变人单核细胞中M1/M2趋化因子的表达。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（Cell and Bioscience, 2022, 12(1). DOI:10.1186/s13578-022-00767-w）
（Cell and Bioscience, 2022, 12(1). DOI:10.1186/s13578-022-00767-w）
TSLP regulates mitochondrial ROS-induced mitophagy via histone modification in human monocytes
Lin YC, Lin YC, Tsai ML, et al.
Abstract
BACKGROUNDS：Thymic stromal lymphopoietin (TSLP) is a Th2-like cytokine involved in asthma pathogenesis. Excessive reactive oxygen species (ROS) production can lead to airway inflammation, hyperresponsiveness and remodeling. Mitophagy, followed by ROS production, is the selective degradation of mitochondria by autophagy and often occurs in defective mitochondria.
OBJECTIVE：In the present study, we aimed to examine the effects of TSLP on ROS production and mitophagy in human monocytes and to investigate the underlying mechanisms, including epigenetic regulation.
RESULTS：TSLP induced ROS generation, and the effects were reversed by the antioxidant N-acetylcysteine (NAC) in THP-1 cells. Transmission electron microscopy images showed donut-shaped mitochondria that lost the cristae ultrastructure after TSLP stimulation. A decrease in mitochondrial membrane potential, decreased MTCO2 expression, and increased mitochondrial DNA release after TSLP stimulation were found. TSLP enhanced mitochondrial complex I and complex II/III activity and increased mitochondrial copy numbers and the expression of the complex II SHDA gene. TSLP-induced SHDA expression was inhibited by the histone acetyltransferase inhibitor anacardic acid (AA) and the histone methyltransferase inhibitor methylthioadenosine (MTA), and chromatin immunoprecipitation assays revealed that TSLP enhanced H3 acetylation, H4 acetylation, and H3K4 and H3K36 trimethylation in the SHDA promoter. Confocal laser microscopy showed that TSLP treatment increased the signals of the mitophagy-related proteins PINK1, LC3, phospho-parkin and phospho-ubiquitin, and pretreatment with AA and MTA reduced TSLP-induced PINK1 and LC3 accumulation in mitochondria. Western blot analysis showed that TSLP significantly increased phosphor-AMPK signal intensity, and the effects were inhibited by the antioxidant NAC. The increased signal intensities of the mitophagy-related proteins PINK1, Parkin and LC3 I/II were decreased by dorsomorphin, an AMPK inhibitor. TSLP decreased M1-related cytokine CXCL-10 production and increased M2-related cytokine CCL-1 and CCL-22 production, which was suppressed by the mitophagy inhibitor Mdivi-1 and PINK1 gene knockdown.
CONCLUSIONS：Epithelial-derived TSLP regulates ROS production and mitophagy through AMPK activation and histone modification and alters M1/M2 chemokine expression in human monocytes.
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