NF-κB通过TRPM7通道蛋白调控心脏纤维化的机制研究

Mechanism of NF-κB Regulating Cardiac Fibrosis through TRPM7 Channel Protein

探索NF-κB和TRPM7之间的相互关系,及其在心脏纤维化过程中的具体调控机制。本研究从细胞和动物层面分别构建了纤维化模型(TGF-β处理MEF细胞、ISO处理小鼠),并利用siRNA干扰和黄芪甲苷靶向抑制TRPM7,通过RT-qPCR和Western Blot分别检测纤维化因子(MMP9和α-SMA)以及TRPM7在mRNA和蛋白水平的表达。同时采用ELISA法测定细胞上清、小鼠和心衰患者血清中的NF-κB的浓度。进一步采用生物信息学方法分析TRPM7启动子区域转录调控信息,预测该区域与NF-κB信号通路的相关性。无论在细胞还是动物模型中,与对照组相比,纤维化组TRPM7的表达显著增加(P<0.05),NF-κB的浓度也显著升高(P<0.05),当TRPM7的表达被抑制后,心脏纤维化程度得到明显的缓解。生物信息学分析表明NF-κB的主要组分NF-κB p65可能通过与TRPM7启动子特定区域结合从而调节TRPM7的表达,最终起到调控心脏纤维化的作用。TRPM7通道蛋白在心脏纤维化过程中发挥着关键作用,NF-κB的上调与纤维化因子的激活关系密切,NF-κB可能是TRPM7通道蛋白的直接转录调控因子,其通过促进TRPM7表达调控心脏纤维化。

To explore the relationship between NF-κB and TRPM7 and the specific regulatory mechanism in cardiac fibrosis,in this study,fibrosis models(TGF-βtreated MEF cells and ISO-treated mice)were constructed at the cellular and animal levels,respectively.TRPM7 was inhibited by siRNA interference and astragaloside IV targeting.The mRNA and protein expressions of fibrosis factor(MMP9 andα-SMA)and TRPM7 were detected by RT-qPCR and Western Blot,respectively.The concentration of NF-κB in cell supernatant,serum of mice and patients with heart failure was determined by ELISA.The transcriptional regulation information of TRPM7 promoter region was analyzed by bioinformatics method,and the correlation between this region and NF-κB signaling pathway was predicted.In both cellular and animal models,TRPM7 expression was significantly increased in the fibrosis group compared to the control group(P<0.05),the concentration of NF-κB was also significantly increased(P<0.05),when the expression of TRPM7 was inhibited,the degree of cardiac fibrosis was significantly relieved.Bioinformatics analysis indicated that NF-κB p65,a major component of NF-κB,may regulate the expression of TRPM7 by binding to the specific region of TRPM7 promoter,and ultimately play a role in regulating cardiac fibrosis.TRPM7 channel protein plays a key role in the process of cardiac fibrosis.The upregulation of NF-κB is closely related to the activation of fibrosis factor.NF-κB may be a direct transcriptional regulator of TRPM7 channel protein,and it regulates cardiac fibrosis by promoting the expression of TRPM7.