Inhibition of ERK 1/2 pathway downregulates YAP1/TAZ signaling in human cardiomyocytes exposed to hyperglycemic conditions
Inhibition of ERK 1/2 pathway downregulates YAP1/TAZ signaling in human cardiomyocytes exposed to hyperglycemic conditions
复制标题
抑制 ERK 1/2 通路可下调暴露于高血糖条件下的人心肌细胞中的 YAP1/TAZ 信号传导
DOI：
10.1016/j.bbrc.2023.01.014
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发表时间：
2023
影响因子：
3.1
通讯作者：
Singh, Irtisha
中科院分区：
文献类型：
--
作者：
Joddar, Binata;Loyola, Carla D.;Ramirez, Salma P.;Muruganandham, Abhinaya;Singh, Irtisha
高血糖介导的心功能不全是血管并发症发展的急性启动剂，导致心脏纤维化。为了研究高血糖介导的心肌细胞变化的影响，分别在正常血糖（5 mM或25 mM D-葡萄糖）和高血糖（5 → 50 mM或25 → 50 mM D-葡萄糖）条件下体外培养细胞。高血糖暴露24小时后，收集细胞进行RNA测序（RNA-seq）研究，以进一步研究高血糖条件下培养的样本中与炎症和纤维化相关的差异表达基因（DEG），并与正常血糖条件下培养的样本进行比较。进行蛋白质印迹以评估在高血糖诱导的应激条件下YAP 1/TAZ的蛋白质表达，因为已知其参与纤维化和血管炎症介导的条件。RNA-seq揭示了多个靶点的DEG，包括基质金属蛋白酶和炎症介质，与25 → 50 mM条件相比，5 → 50 mM条件下其表达显著改变。蛋白质印迹显示在这些条件下YAP 1/TAZ途径的蛋白质表达也显著上调（5 → 50 mM）。为了进一步探索炎性细胞外信号调节激酶（ERK 1/2）及其下游对YAP 1/TAZ表达的影响之间的关系，我们研究了在5 → 50 mM条件下抑制ERK 1/2信号级联的影响。ERK 1/2抑制剂的应用在5 → 50 mM条件下抑制了YAP 1/TAZ蛋白的表达，这种策略可能有助于预防和改善高血糖相关的心血管损伤和炎症。
Hyperglycemia-mediated cardiac dysfunction is an acute initiator in the development of vascular complications, leading to cardiac fibrosis. To investigate the effects of hyperglycemia-mediated changes in cardiomyocytes, cells were cultured in-vitro under normoglycemic (5 mM or 25 mM D-glucose) and hyperglycemic (5 → 50 mM or 25 → 50 mM D-glucose) conditions, respectively. After 24-h of hyperglycemic exposure, cells were collected for RNA-sequencing (RNA-seq) studies to further investigate the differentially expressed genes (DEG) related to inflammation and fibrosis in samples cultured under hyperglycemic-in comparison with normoglycemic-conditions. Western Blotting was done to evaluate the protein expression of YAP1/TAZ under hyperglycemia induced stress conditions, as it is known to be involved in fibrotic and vascular inflammatory-mediated conditions. RNA-seq revealed the DEG of multiple targets including matrix metalloproteinases and inflammatory mediators, whose expression was significantly altered in the 5 → 50 mM in comparison with the 25 → 50 mM condition. Western Blotting showed a significant upregulation of the protein expression of the YAP1/TAZ pathway under these conditions as well (5 → 50 mM). To further probe the relationship between the inflammatory extracellular-signal-regulated kinase (ERK 1/2) and its downstream effects on YAP1/TAZ expression we studied the effect of inhibition of the ERK 1/2 signaling cascade in the 5 → 50 mM condition. The application of an ERK 1/2 inhibitor inhibited the expression of the YAP1/TAZ protein in the 5 → 50 mM condition, and this strategy may be useful in preventing and improving hyperglycemia associated cardiovascular damage and inflammation.