ROCK-dependent ATP5D modulation contributes to the protection of notoginsenoside NR1 against ischemia-reperfusion-induced myocardial injury

Abstract

Cardiac ischemia-reperfusion (I/R) injury remains a challenge for clinicians, which initiates with energy metabolism disorder. The present study was designed to investigate the protective effect of notoginsenoside R1 (NR1) on I/R-induced cardiac injury and underlying mechanism. Male Sprague-Dawley rats were subjected to 30-min occlusion of the left coronary anterior descending artery followed by reperfusion with or without NR1 pretreatment (5 mg·kg(-1)·h(-1)). In vitro, H9c2 cells were cultured under oxygen and glucose deprivation/reoxygenation conditions after NR1 (0.1 mM), Rho kinase (ROCK) inhibitor Y-27632 (10 μM), or RhoA/ROCK activator U-46619 (10 nM) administration. Myocardial infarct size, myocardial histology, and cardiac function were evaluated. Myofibril and mitochondria morphology were observed by transmission electron microscopy. F-actin and apoptosis were determined by immunofluorescence and TUNEL staining. ATP and AMP content were assessed by ELISA. Phosphorylated-AMP-activated protein kinase, ATP synthase subunits, apoptosis-related molecules, and the level and activity of ROCK were determined by Western blot analysis. We found that NR1 pretreatment ameliorated myocardial infarction, histological injury, and cardiac function induced by I/R. Furthermore, similar to the effect of Y-27632, NR1 improved H9c2 cell viability, maintained actin skeleton and mitochondria morphology, and attenuated apoptosis induced by oxygen and glucose deprivation/reoxygenation. Importantly, NR1 prevented energy abnormity, inhibited the expression and activation of ROCK, and restored the expression of the mitochondrial ATP synthase δ-subunit both in vivo and in vitro, whereas U-46619 suppressed the effect of NR1. These results prove NR1 as an agent able to prevent I/R-induced energy metabolism disorder via inhibiting ROCK and enhancing mitochondrial ATP synthase δ-subunits, which at least partially contributes to its protection against cardiac I/R injury.

Panax notoginseng saponins provide neuroprotection by regulating NgR1/RhoA/ROCK2 pathway expression, in vitro and in vivo

“…The combination of PNS and NEP1-40 showed no obvious synergistic effect. A recent study showed that one effective constituent of PNS, notoginsenoside NR1, prevents I/R-induced myocardial injury by inhibiting ROCK (He et al, 2014), suggesting that the effective constituent against ROCK2 in ischemic stroke might be notoginsenoside NR1. However, further studies are needed to confirm this hypothesis, as well as to determine the mechanisms of action of PNS on NgR1 and RhoA expression following cerebral ischemia injury.…”

Ginsenoside Rb1 protects against ischemia/reperfusion-induced myocardial injury via energy metabolism regulation mediated by RhoA signaling pathway

“…ATPase is a multisubunit complex consisting of 15 subunits including ATP 5D in mammals, which integrate into two domains, F1 and Fo. ATPase is reported to respond to ischemia and reperfusion by changing the expression of subunit protein (s)45 leading to ATPase dysfunction. ATPase dysfunction may impose dual deleterious impacts on the tissue underwent I/R, which, on one hand, results in ATP depletion thus affects all the energy consuming processes in the cell, such as cell contraction and ion pumps, as well as triggers the production of reactive oxidative species, and disrupts the mitochondrial membrane potential (ΔΨm), on the other hand, is implicated in the progressing of apoptosis6.…”

“…QiShenYiQi Pills ® (a compound Chinese medicine containing Rb1 as a major component) has proven able to regulate energy metabolism deficiency and expression of ATP 5D in cardiac I/R injury1213. Our previous study reported that notoginsenoside R1 (NR1) ameliorated I/R induced myocardial injury by regulating levels of ROCK1 and ATP5D, implying a likely link between RhoA signaling pathway and energy metabolism regulation4. However, it is so far unclear whether Rb1 can improve I/R-induced cardiac energy deficiency and, if yes, what is the role of RhoA signaling pathway in the process.…”

Myocardial Ischemia-Reperfusion Injury: The Perioperative Application and Cardiac Protective Potential of Ginsenoside Preparations

“…Notoginsenoside R1 markedly ameliorated myocardial infarction, improved cardiac function and increased cell viability. More importantly, notoginsenoside R1 prevented energy metabolism abnormity both in in vivo and in vitro models, mainly by down-regulating the expression of Rho kinase (ROCK) and up-regulating the expression of the mitochondrial ATP synthase δ-subunit (63). All these results indicate that a variety of ginsenosides can improve energy metabolism and alleviate the degree of reperfusion injury.…”