Elsevier

Free Radical Biology and Medicine

Volume 97, August 2016, Pages 489-500
Free Radical Biology and Medicine

Mitochondrial Peroxiredoxin-3 protects against hyperglycemia induced myocardial damage in Diabetic cardiomyopathy

https://doi.org/10.1016/j.freeradbiomed.2016.06.019Get rights and content

Highlights

  • Expression of Prx-3 was decreased in high glucose treated H9c2 cells and STZ induced diabetic rats.

  • Prx-3 induction reduced oxidative stress, apoptosis, hypertrophy and fibrosis.

  • Prx-3 induction prevented contractile dysfunction in diabetic cardiomyopathy.

Abstract

Mitochondrial oxidative stress has emerged as a key contributor towards the development of diabetic cardiomyopathy. Peroxiredoxin-3 (Prx-3), a mitochondrial antioxidant, scavenges H2O2 and offers protection against ROS related pathologies. We observed a decrease in the expression of Prx-3 in the hearts of streptozotocin (STZ) induced diabetic rats, and also high glucose treated H9c2 cardiac cells, which may augment oxidative stress mediated damage. Hence we hypothesized that overexpression of Prx-3 could prevent the cardiac damage associated with diabetes. In this study we used quercetin (QUE) to achieve Prx-3 induction in vivo, while a Prx-3 overexpressing H9c2 cell line was employed for carrying out in vitro studies. Diabetes was induced in Wistar rats by a single intraperitoneal injection of STZ. Quercetin (50 mg/kg body weight) was delivered orally to hyperglycemic and age matched control rats for 2 months. Quercetin treatment induced the myocardial expression of Prx-3 but not Prx-5 both in control and STZ rats. Prx-3 induction by quercetin prevented diabetes induced oxidative stress as confirmed by decrease in expression of markers such as 4-HNE and mitochondrial uncoupling protein, UCP-3. It was also successful in reducing cardiac cell apoptosis, hypertrophy and fibrosis leading to amelioration of cardiac contractility defects. Overexpression of Prx-3 in cultured H9c2 cardiac cells could significantly diminish high glucose inflicted mitochondrial oxidative damage and apoptosis, thus strengthening our hypothesis. These results suggest that diabetes induced cardiomyopathy can be prevented by elevating Prx-3 levels thereby providing extensive protection to the diabetic heart.

Introduction

Sedentary lifestyles have increased the occurrence of obesity, thus propelling the incidence of diabetes to reach an epidemic level. Statistical projections predict that over 300 million people will develop diabetes by 2025 [1]. Diabetic cardiomyopathy (DCM) is an independent risk factor, which impairs myocardial performance and is a key manifestation of diabetic condition [2]. Pathogenesis of DCM is multifactorial, but underlying mechanism is partially understood [3], [4], [5], [6]. Among the possible molecular mechanisms responsible for the progression of diabetic cardiomyopathy, mounting evidence from curated studies have intimately connected the generation of reactive oxygen species (ROS) and incidence of apoptosis to this state [7], [8].

Mitochondria are principle source of ROS under hyperglycemic conditions [9]. Most of the superoxide (O2∙−), produced at electron transport chain (ETC) [10], is dismutated under physiological conditions by mitochondrial Manganese superoxide dismutase (MnSOD) to form hydrogen peroxide (H2O2). Even though, MnSOD relieves mitochondrial oxidative stress caused by O2∙− [11] it further enhances a different type of oxidative stress. H2O2 can damage cellular macromolecules such as proteins, lipids, and nucleic acids, especially after its conversion to hydroxyl radical (OH∙) by Haber-Weiss reaction. Recently, our laboratory has demonstrated that Monoamine Oxidase-A (MAO-A), present in the outer mitochondrial membrane is also an important source of H2O2 and involved in development of DCM [12]. Based on these results, mitochondrial antioxidants are projected to be the first line-of-defense mechanism against ROS generation in the mitochondria and thus, may improve the myocardial performance in diabetes mellitus. Mitochondrial H2O2 can be decomposed by Glutathione peroxidases (GPxs) 1, 4 and Peroxiredoxins (Prxs) 3, 5. However, an absence of catalase in mitochondria of myocytes [13] and reduced scavenging of H2O2 by GPx1 [14] highlights the importance of Peroxiredoxins in removal of mitochondrial H2O2. Prxs are thiol-dependent antioxidants which reduce H2O2 at its cysteine residues containing active sites. Prxs are present in six isoforms Prx-1 to -6. Of these, Prx-3 contains mitochondrial localization sequence and exclusively found in mitochondria. Prx-5 is also linked with mitochondria in addition to nucleus and peroxisomes. The ability of Prx-3 and Prx-5 as active antioxidants depends on their recycling by the mitochondrial electron donor Thioredoxin-2 (Trx-2) complex [15], [16]. The reduced form of Trx-2 is then regenerated by Thioredoxin reductase-2 (TrxR2) at the expense of NADPH [17], [18], [19]. Trx-2 and TrxR2 reside in the mitochondrial matrix and operate independently from the cytosolic Trx network. According to kinetic studies, Prx-3 has emerged as a principle scavenger of H2O2 in mitochondria [20]. The greater efficiency of mitochondrial Prx-3 and -5 together with Trx-2, TrxR2 and NADPH, may attribute to protection against high glucose induced oxidative stress. Infact, overexpression of Prx-3 has been reported to prevent the left ventricular remodeling after myocardial infarction in transgenic mice [21], and Prx-3 also has crucial role in contractile function of skeletal muscle by regulating mitochondrial homeostasis [22]. Prx-5 overexpression protects mitochondrial DNA damage induced by H2O2 [23] and also human tendon cells against apoptosis and loss of cellular function during oxidative stress [24]. However, the role of mitochondrial Prx in DCM prevention has not been explored to its full potential.

In the current study, we found that Prx-3 but not Prx-5 expression was significantly reduced in the heart of diabetic rats. Therefore, the aim of present study was to examine the effect of Prx-3 induction on oxidative stress induced myocardial damage in diabetic condition.

Section snippets

In vivo animal model

Male Wistar rats were bred and maintained at animal house facility of National Centre for Cell Science. All animal experiments were duly approved by the Institutional Animal Ethics Committee (IAEC) under reference number IAEC/2012/B-195 dated on 8/9/12 of National Centre for Cell Science and were performed in full compliance of the extant guidelines and principles. Food and water were available ad libitum. Diabetes was induced at 6–8 weeks of age by single intraperitoneal (IP) injection of

Prx-3 expression is down regulated in cardiac cells under hyperglycemic condition

Excessive oxidative stress, induced by ROS and RNS generation is associated with diabetic cardiomyopathy [31]. To determine whether mitochondrial antioxidants Prx-3, Prx-5 and their electron donor Trx-2 are involved in the regulation of hyperglycemia induced oxidative stress, we examined their expression levels in both in vitro and in vivo models. H9c2 cells, at 60% confluency, were subjected to normal and high glucose for 24, 48, 72 and 96 h. Expression of Prx-3 and Trx-2 were found to be

Discussion

The current study demonstrates that overexpression of the mitochondrial antioxidant; Prx-3, offers enhanced protection against diabetes-induced cardiac injury. We observed that Prx-3 induction by quercetin treatment prevented a multitude of cardiac complications such as contractility defects, hypertrophy, myocardial fibrosis and apoptosis in STZ induced diabetic rats. Under in vitro conditions too, beneficial effects of Prx-3 gene overexpression were found to be associated with an attenuation

Authors’ contributions

S.A. and S.L.S. contributed to the design of the study, data analysis, and interpretation of results. S.A., P.U., S.S., contributed to data acquisition. S.A., P.U., S.S., and S.L.S. contributed to drafting of the manuscript.

Conflicts of interests

The authors declare that they have no conflicts of interest associated with this manuscript.

Sources of funding

This work was supported by intramural funding of National Centre for Cell Science, Department of Biotechnology, India. SA (Sr. No. 2061030780) PU (Sr. No. 2061030917) and SS (Sr. No. 2061030712) and are recipients of University Grants Commission (UGC) Senior Research Fellowship.

Acknowledgments

We thank Dr. S.C. Mande, Director, National Centre for Cell Science (Pune, India) for encouragement and support. We express thanks to Aparajita Dasgupta for proofreading the manuscript. We also acknowledge help from the staff of the experimental animal facility at the National Centre for Cell Science.

References (43)

  • B. Kalyanaraman et al.

    Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations

    Free Radic. Biol. Med.

    (2012)
  • H.J. Forman et al.

    Even free radicals should follow some rules: a guide to free radical research terminology and methodology

    Free Radic. Biol. Med.

    (2015)
  • S.G. Rhee et al.

    Peroxiredoxins: a historical overview and speculative preview of novel mechanisms and emerging concepts in cell signaling

    Free Radic. Biol. Med.

    (2005)
  • E. Schroder et al.

    Hydrogen peroxide as an endogenous mediator and exogenous tool in cardiovascular research: issues and considerations

    Curr. Opin. Pharmacol.

    (2008)
  • H. King et al.

    Global burden of diabetes, 1995–2025: prevalence, numerical estimates, and projections

    Diabetes Care

    (1998)
  • S. Rubler et al.

    Noninvasive estimation of myocardial performance in patients with diabetes. Effect of alcohol administration

    Diabetes

    (1978)
  • R.B. Devereux et al.

    Impact of diabetes on cardiac structure and function: the strong heart study

    Circulation

    (2000)
  • G.S. Francis

    Diabetic cardiomyopathy: fact or fiction?

    Heart

    (2001)
  • L. Cai et al.

    Oxidative stress and diabetic cardiomyopathy: a brief review

    Cardiovasc. Toxicol.

    (2001)
  • G.Z. Feuerstein et al.

    Apoptosis in cardiac diseases: stress- and mitogen-activated signaling pathways

    Cardiovasc. Res.

    (2000)
  • T. Nishikawa et al.

    Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage

    Nature

    (2000)
  • Cited by (41)

    • The m6A reader YTHDF3-mediated PRDX3 translation alleviates liver fibrosis

      2022, Redox Biology
      Citation Excerpt :

      Recent studies have shown that PRDX3 protects cells from excess mitochondrial ROS accumulation by eliminating approximately 90% of mitochondrial H2O27. For instance, the overexpression of PRDX3 reportedly prevents mitochondrial oxidative stress-induced damage in a variety of diseases, such as chronic kidney injury and diabetes [8,9]. Moreover, PRDX3 exerts hepatoprotective effects on nonalcoholic fatty liver disease, alcoholic liver injury and acetaminophen-induced liver injury [10–12].

    • Therapeutic potential of targeting oxidative stress in diabetic cardiomyopathy

      2021, Free Radical Biology and Medicine
      Citation Excerpt :

      Interestingly, induction of PRX3 expression by the flavonoid, quercetin, prevented T1D-induced oxidative stress and cardiac dysfunction [181]. Furthermore, HG-induced mitochondrial oxidative damage was reduced following overexpression PRX3 in cultured cardiomyocytes [181]. When insufficient amounts of reduced PRX are available to deal with peroxides, hyperoxidation of PRX may occur (Prx-SO2/3H), causing inactivation of the peroxidase activity [188].

    • Therapeutic approaches to diabetic cardiomyopathy: Targeting the antioxidant pathway

      2020, Prostaglandins and Other Lipid Mediators
      Citation Excerpt :

      Prx-3 levels were reduced in rat H9C2 cardiac cells maintained under hyperglycaemic conditions, and also in STZ-induced diabetic rats. Induction of Prx-3 with Quercetin prevented diabetes-induced oxidative stress [259]. Rutin: Rutin is a glycoside of quercetin, that exhibits anti-oxidant and anti-inflammatory properties.

    View all citing articles on Scopus
    1

    Contributed equally to this work.

    View full text