The GEO database and RNA sequencing were used to predict downstream gene. TTC staining was used to visualize the myocardial infarction area. Masson staining was used to assess the level of fibrosis. IF was used to examine the expression of SHP2, CTGF, ROS. RT-PCR analysis was used to quantify the expression of SHP2 mRNA. Western blot was used to detect the protein expression levels of SHP2, p-PERK, MFN1, NLRP3, NOX2, and NOX3.
GEO and transcriptomic data revealed low expression of SHP2 in the heart tissues I/R mice. In the I/R mouse model, TTC staining result showed that allicin can reduce the area of myocardial infarction; Masson staining results indicated that allicin can reduce fibrosis; Macrophage transcriptome sequencing found SHP2 is a target gene of allicin; Immunofluorescence showed allicin can increase SHP2; qPCR results showed allicin can raise SHP2 mRNA level; Immunofluorescence indicated that allicin can inhibit ROS in myocardial infarction tissue, but the specific SHP2-KD eliminates changes in ROS. Western blot analysis demonstrated allicin can increase SHP2 protein and reduce the expression of p-PERK, MFN1, NLRP3, NOX2, and NOX3; SHP2-KD eliminates the expression differences in p-PERK, MFN1, NLRP3, NOX2, and NOX3.
Allicin can modulate p-PERK activation by enhancing the expression of SHP2, thereby inhibiting myocardial ischemia-reperfusion-induced oxidative stress in mice.
方法:使用GEO数据库和RNA测序来预测下游基因。用TTC染色观察心肌梗死面积。Masson染色用于评估纤维化水平。IF用于检测SHP2,CTGF的表达,ROS。RT-PCR分析用于定量SHP2mRNA的表达。Westernblot检测SHP2、p-PERK、MFN1、NLRP3、NOX2和NOX3。
结果:GEO和转录组数据显示SHP2在I/R小鼠心脏组织中的低表达。在I/R鼠标模型中,TTC染色结果显示大蒜素可以减少心肌梗死面积;Masson染色结果显示大蒜素可以减少纤维化;巨噬细胞转录组测序发现SHP2是大蒜素的靶基因;免疫荧光显示大蒜素可以增加SHP2;qPCR结果显示大蒜素可以提高SHP2的mRNA水平;免疫荧光显示大蒜素可以抑制心肌梗死组织中的ROS。但是特定的SHP2-KD消除了ROS的变化。Westernblot分析表明,大蒜素能增加SHP2蛋白的表达,降低p-PERK的表达,MFN1、NLRP3、NOX2和NOX3;SHP2-KD消除了p-PERK的表达差异,MFN1、NLRP3、NOX2和NOX3。
结论:大蒜素可以通过增强SHP2的表达来调节p-PERK的激活,从而抑制小鼠心肌缺血再灌注诱导的氧化应激。