PDF(Pubmed)
Abstract：
UNASSIGNED: Doxorubicin (Dox) can induce cardiotoxicity, thereby restricting the utility of this potent drug. Herein, the study ascertained the mechanism of the N6-methyladenosine (m6A) demethylase fat mass and obesity-associated protein (FTO) in pyroptosis and inflammation during Dox-induced heart failure (HF).
UNASSIGNED: Serum samples were collected from HF patients for detection of the expression of FTO and toll-like receptor 4 (TLR4). Dox-treated H9C2 cardiomyocytes were chosen for in vitro HF modeling, followed by measurement of FTO and TLR4 expression. Cardiomyocytes were detected for viability, apoptosis, spatial distribution of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3), and the levels of lactic dehydrogenase, inflammatory factors, oxidative stress markers, and pyroptosis-related proteins. The m6A levels of mRNA were examined. RNA immunoprecipitation (RIP) and mRNA stability measurement were used to determine mRNA and protein expression, and RNA m6A dot blot and methylated-RIP assay were performed to detect m6A methylation levels. The expression of p-NF-κB p65 and p-IκB-α was measured by western blotting.
UNASSIGNED: In the serum of HF patients, FTO was elevated while TLR4 was decreased. Dox treatment reduced FTO expression and increased m6A methylation levels and TLR4 expression in H9C2 cells. Overexpression of FTO and knockdown of TLR4 reduced apoptosis, cytotoxicity, inflammation, pyroptosis, oxidative stress, NLRP3 co-localization, and fluorescence intensity in Dox-induced H9C2 cells. Mechanistically, FTO resulted in reduced binding activity of YTHDF1 to TLR4 mRNA via m6A demethylation of TLR4, thus declining TLR4, p-NF-κB p65, and p-IκB-α expression. TLR4 knockdown counteracted the effects of FTO knockdown on Dox-induced H9C2 cells.
UNASSIGNED: FTO alleviated Dox-induced HF by blocking the TLR4/NF-κB pathway.
UNASSIGNED: Serum samples were collected from HF patients for detection of the expression of FTO and toll-like receptor 4 (TLR4). Dox-treated H9C2 cardiomyocytes were chosen for in vitro HF modeling, followed by measurement of FTO and TLR4 expression. Cardiomyocytes were detected for viability, apoptosis, spatial distribution of NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3), and the levels of lactic dehydrogenase, inflammatory factors, oxidative stress markers, and pyroptosis-related proteins. The m6A levels of mRNA were examined. RNA immunoprecipitation (RIP) and mRNA stability measurement were used to determine mRNA and protein expression, and RNA m6A dot blot and methylated-RIP assay were performed to detect m6A methylation levels. The expression of p-NF-κB p65 and p-IκB-α was measured by western blotting.
UNASSIGNED: In the serum of HF patients, FTO was elevated while TLR4 was decreased. Dox treatment reduced FTO expression and increased m6A methylation levels and TLR4 expression in H9C2 cells. Overexpression of FTO and knockdown of TLR4 reduced apoptosis, cytotoxicity, inflammation, pyroptosis, oxidative stress, NLRP3 co-localization, and fluorescence intensity in Dox-induced H9C2 cells. Mechanistically, FTO resulted in reduced binding activity of YTHDF1 to TLR4 mRNA via m6A demethylation of TLR4, thus declining TLR4, p-NF-κB p65, and p-IκB-α expression. TLR4 knockdown counteracted the effects of FTO knockdown on Dox-induced H9C2 cells.
UNASSIGNED: FTO alleviated Dox-induced HF by blocking the TLR4/NF-κB pathway.
摘要：
■阿霉素（Dox）可以诱导心脏毒性，从而限制了这种强效药物的效用。在这里，该研究确定了N6-甲基腺苷(m6A)脱甲基酶脂肪量和肥胖相关蛋白(FTO)在Dox诱导的心力衰竭(HF)期间的焦亡和炎症中的作用机制.
从HF患者收集血清样品用于检测FTO和toll样受体4(TLR4)的表达。选择Dox处理的H9C2心肌细胞进行体外HF建模，然后测量FTO和TLR4的表达。检测心肌细胞的活力，凋亡，NOD-的空间分布，含LRR和Pyrin结构域的蛋白3（NLRP3），和乳酸脱氢酶的水平，炎症因子，氧化应激标志物，和焦亡相关蛋白。检测mRNA的m6A水平。RNA免疫沉淀(RIP)和mRNA稳定性测量用于确定mRNA和蛋白质表达,和RNAm6A斑点印迹和甲基化RIP测定以检测m6A甲基化水平。免疫印迹法检测p-NF-κBp65和p-IκB-α的表达。
■在HF患者的血清中，FTO升高而TLR4降低。Dox处理降低了FTO表达并增加了H9C2细胞中的m6A甲基化水平和TLR4表达。FTO的过表达和TLR4的敲除减少细胞凋亡，细胞毒性，炎症,焦亡,氧化应激，NLRP3共同本地化，和Dox诱导的H9C2细胞中的荧光强度。机械上,FTO通过TLR4的m6A去甲基化导致YTHDF1与TLR4mRNA的结合活性降低，从而降低了TLR4，p-NF-κBp65和p-IκB-α的表达。TLR4敲低抵消FTO敲低对Dox诱导的H9C2细胞的影响。
■FTO通过阻断TLR4/NF-κB途径减轻Dox诱导的HF。
从HF患者收集血清样品用于检测FTO和toll样受体4(TLR4)的表达。选择Dox处理的H9C2心肌细胞进行体外HF建模，然后测量FTO和TLR4的表达。检测心肌细胞的活力，凋亡，NOD-的空间分布，含LRR和Pyrin结构域的蛋白3（NLRP3），和乳酸脱氢酶的水平，炎症因子，氧化应激标志物，和焦亡相关蛋白。检测mRNA的m6A水平。RNA免疫沉淀(RIP)和mRNA稳定性测量用于确定mRNA和蛋白质表达,和RNAm6A斑点印迹和甲基化RIP测定以检测m6A甲基化水平。免疫印迹法检测p-NF-κBp65和p-IκB-α的表达。
■在HF患者的血清中，FTO升高而TLR4降低。Dox处理降低了FTO表达并增加了H9C2细胞中的m6A甲基化水平和TLR4表达。FTO的过表达和TLR4的敲除减少细胞凋亡，细胞毒性，炎症,焦亡,氧化应激，NLRP3共同本地化，和Dox诱导的H9C2细胞中的荧光强度。机械上,FTO通过TLR4的m6A去甲基化导致YTHDF1与TLR4mRNA的结合活性降低，从而降低了TLR4，p-NF-κBp65和p-IκB-α的表达。TLR4敲低抵消FTO敲低对Dox诱导的H9C2细胞的影响。
■FTO通过阻断TLR4/NF-κB途径减轻Dox诱导的HF。
