Allicin protects against myocardial I/R by accelerating angiogenesis via the miR-19a-3p/PI3K/AKT axis

Abstract

Objectives: Allicin is an allyl 2-propenethiosulfinate or diallyl thiosulfinate acid with cardioprotective effects in myocardial ischemia/reperfusion (MI/R) injury. This study aims to examine the underlying mechanism by which Allicin protects against MI/R. Methods: C57BL6 mice were subjected to either sham or MI/R surgery, and mice in the Allicin group were injected with Allicin (5 mg/ml) before the induction of ischemia. The cardiac function and histopathology of experimental mice were evaluated by ultrasound quantification and Masson staining. We next measured the capillary angiogenesis of the peri-infarct area by Masson staining and immunohistochemical staining. The miRNA microarray was carried out to examine the expressed miRNAs in MI/R tissues and corresponding normal tissues. Real-time quantitative polymerase chain reaction (q-PCR) was performed to validate the selected miRNA-19α-3p gene expression. Besides, we evaluated the myocardial lactate dehydrogenase and COX-2 by immunofluorescence staining. The western blot analysis was used to evaluate the protein levels of p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein in the Allicin and Model group. In vitro study, LPS stimulated Tie2 expressing macrophages were cultured in an ischemic buffer. We evaluated the accumulation of VEGF by fura-2/AM fluorescence. Besides, Western blotting was performed to examine the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2. The PI3K inhibitor was applied to investigate whether Allicin-induced myocardial ischemia-reperfusion injury protection is mediated via the PI3K/AKT pathway. And the miR-19α-3p mimic/inhibitor were transfected to promote/inhibit the expression of miR-19a-3p for verifying the regulation of miR-19a-3p on PI3K pathway. Results: Allicin pretreatment significantly improved I/R-induced cardiac function damage. Furthermore, Allicin could repress cardiac fibrosis, as evidenced by reduced areas of cardiac fibrosis. Allicin’s effect on the MI/R was associated with increased capillary angiogenesis. Microarray analysis exposed that miR-19a-3p down-regulated PIK3CA (PI3K) expression by directly targeting the PIK3CA gene. The regulation of the angiogenesis pathway and gene miRNA-19a-3p might affect the Allicin-induced MI/R protection. Immunofluorescence staining revealed that COX-2 and myocardial lactate dehydrogenase were significantly increased after Allicin treatment. Furthermore, western blot analysis demonstrated that p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein levels were also increased in the Allicin group. In vitro study, the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2 were significantly increased in the Allicin-treated Tie2 expressing macrophages. These effects were partially reversed by PI3K inhibitor (Wortmannin) treatment. MiR-19α-3p plays an important role in myocardial I/R injury. It could regulate the activity of the PI3K-AKT pathway. And inhibition of miR-19a-3p promoted angiogenesis by regulating P...

Blockage of Fibronectin 1 Ameliorates Myocardial Ischemia/Reperfusion Injury in Association with Activation of AMP-LKB1-AMPK Signaling Pathway

“…Furthermore, inflammatory cytokines can stimulate myofibroblast activation and cause excessive collagen accumulation resulting in cardiac fibrosis [ 36 , 37 ]. Interestingly, several studies show that short-time I/R (2-24 h) can significantly increase expression of collagen I/III (1.5-fold) and fibrotic area (about 15-25-fold) in the myocardial tissues of rats or mice compared with sham control [ 38 – 40 ], suggesting that increased collagen production is an early event during myocardial I/RI. Consistent with these data, our results further confirmed that I/R for 24 and 72 h also markedly increased expression of cardiac collagen I, collagen III, and ɑ-SMA and fibrotic area compared with sham control, and this effects were greatly inhibited in RGDS-treated hearts (Figures 4(d) and 4(e) ).…”

Cellular Mechanisms Underlying the Cardioprotective Role of Allicin on Cardiovascular Diseases

“…Moreover, the antioxidant effects of allicin could contribute to increasing the half-life of the vasoactive metabolites such as NO, and therefore to the maintenance of the vasodilatory actions. Additionally, the cardioprotective effects of allicin have been evidenced in the prevention of apoptosis of ischemic areas, as well as the induction of angiogenesis, which could be useful for the treatment of myocardial infarction, one of the first death causes worldwide [ 2 , 3 , 75 , 77 , 79 , 81 ]. The antiapoptotic and angiogenic activities of allicin may be useful to restore blood flow inducing recovery of injured areas and the effects on the modulation of expression and function of ion channels open the possibility of the use of allicin as an antiarrhythmic agent and pro-angiogenic.…”

“…Other studies reported that allicin improved cardiac function, and reduced the myocardial infarct area, fibrosis, degeneration, necrosis and inflammatory cell infiltration in myocardial tissue as shown in experimental models of ischemia-reperfusion injury [ 77 , 79 , 80 , 81 ]. The cardioprotection induced by allicin was mediated by restoring the levels of SOD, CAT, GPx, and MDA in the myocardial tissue of rats [ 79 ].…”

“…On the other hand, allicin down-regulated the expression of iNOS, p-eNOS/eNOS, Cyt-c, caspase-3, and caspase-9, Bax, phosphorylated Jun-amino-terminal kinase (p-JNK), phosphatidylinositol-4; 5-bisphosphate 3-Kinase catalytic subunit alpha PI3KCA(PI3K), and the phosphorylated G protein-coupled receptor kinase 2 (p-GRK2), Ca 2+ /calmodulin-dependent protein kinases (p-CaMKII), phospholipase C-gamma (p-PLC-γ), and inositol 1,4,5-triphosphate receptor (p-IP3R) [ 77 , 79 ]. Other studies in MI/R in mice showed that allicin, through modulation of the miR-19a-3p/PI3K/AKT pathway, can stimulate angiogenesis and thus revascularization of infarcted areas [ 81 ]. Moreover, it has been demonstrated that allicin improved coronary vascular reactivity in MI/R in rats through the production of hydrogen sulfide (H 2 S) and the regulation of Ca 2+ homeostasis [ 80 ].…”

Dihydroartemisinin Attenuates Hypoxia-Induced Pulmonary Hypertension Through the ELAVL2/miR-503/PI3K/AKT Axis

“…Activation of the PI3K/AKT signaling pathway has been shown to contribute to cell proliferation in several diseases. 23,24 In addition, it has also been reported that miR-503 can inhibit MCT-induced PH in rats. 20 Therefore, we explored the potential interactions between miR-503 and PI3K.…”