Kallikrein/Kinin Protects against Myocardial Apoptosis after Ischemia/Reperfusion via Akt-Glycogen Synthase Kinase-3 and Akt-Bad·14-3-3 Signaling Pathways

Abstract

Our previous study has shown that human tissue kallikrein protected against ischemia/reperfusion-induced myocardial injury. In the present study, we investigated the protective role of local kallikrein gene delivery in ischemia/reperfusion-induced cardiomyocyte apoptosis and its signaling mechanisms in promoting cardiomyocyte survival. Adenovirus carrying the human tissue kallikrein gene was delivered locally into the heart using a catheter-based technique. Expression and localization of recombinant human kallikrein in rat myocardium after gene transfer were determined immunohistochemically. Kallikrein gene delivery markedly reduced reperfusion-induced cardiomyocyte apoptosis identified by both in situ nick end-labeling and DNA fragmentation. Delivery of the kallikrein gene increased phosphorylation of Src, Akt, glycogen synthase kinase (GSK)-3␤, and Bad(Ser-136) but reduced caspase-3 activation in rat myocardium after reperfusion. The protective effect of kallikrein on apoptosis and its signaling mediators was blocked by icatibant and dominant-negative Akt, indicating a kinin B2 receptor-Akt-mediated event. Similarly, kinin or transduction of kallikrein in cultured cardiomyocytes promoted cell viability and attenuated apoptosis induced by hypoxia/reoxygenation. The effect of kallikrein on cardiomyocyte survival was blocked by dominant-negative Akt and a constitutively active mutant of GSK-3␤, but it was facilitated by constitutively active Akt, catalytically inactive GSK-3␤, lithium, and caspase-3 inhibitor. Moreover, kallikrein promoted Bad⅐14-3-3 complex formation and inhibited Akt-GSK-3␤-dependent activation of caspase-3, whereas caspase-3 administration caused reduction of the Bad⅐14-3-3 complex, indicating an interaction between Akt-GSK-caspase-3 and Akt-Bad⅐14-3-3 signaling pathways. In conclusion, kallikrein/kinin protects against cardiomyocyte apoptosis in vivo and in vitro via Akt-Bad⅐14-3-3 and Akt-GSK-3␤-caspase-3 signaling pathways.

Tissue kallikrein infusion prevents cardiomyocyte apoptosis, inflammation and ventricular remodeling after myocardial infarction

“…Our previous study showed that local injection of adenovirus carrying the human tissue kallikrein into the heart protects against cardiomyocyte apoptosis after acute myocardial ischemia/reperfusion [19]. However, kallikrein protein infusion has advantages over adenovirus-mediated delivery.…”

Glycogen synthase kinase-3β plays a pro-apoptotic role in β-adrenergic receptor-stimulated apoptosis in adult rat ventricular myocytes: Role of β1 integrins

“…Transgenic mice expressing wild-type GSK-3β in the heart exhibit dramatic impairment of normal post-natal cardiomyocyte growth as well as abnormal cardiac contractile function [16]. The protective effects of Kallikrein/kinin in ischemia/reperfusion-induced apoptosis are suggested to involve Akt-mediated inactivation of GSK-3β [17]. Inhibition of GSK-3β is also shown to inhibit hypoxia/reperfusion-induced cardiac myocyte apoptosis [18].…”

Recombinant adeno-associated virus-mediated human kallikrein gene therapy protects against hypertensive target organ injuries through inhibiting cell apoptosis

“…The beneficial effects of angiotensin-converting enzyme inhibition in hypertension, cardiovascular and renal disease can be partially attributed to kinin accumulation resulting from the activation of the B2 receptor [16,17] . Previous animal studies suggested that kallikrein/kinin protects against myocardial apoptosis after ischemia/reperfusion injury [18,19] . In our study, delivery of the kallikrein gene protected against cardiomyocyte apoptosis in SHRs; therefore, this gene could potentially be used to treat hypertension.…”