Protective effect of salidroside on cardiac apoptosis in mice with chronic intermittent hypoxia

doi:10.1016/j.ijcard.2014.04.132

International Journal of Cardiology

Volume 174, Issue 3, 1 July 2014, Pages 565-573

https://doi.org/10.1016/j.ijcard.2014.04.132 Get rights and content

Abstract

Background

The goal of this study is to determine if salidroside has protective effects on hypoxia-induced cardiac widely dispersed apoptosis in mice with severe sleep apnea model.

Methods

Sixty-four C57BL/6 J mice 5–6 months of age were divided into four groups, i.e. Control group (21% O₂, 24 h per day, 8 weeks, n = 16); Hypoxia group (Hypoxia: 7% O₂ 60 s, 20% O₂ alternating 60 s, 8 h per day, 8 weeks, n = 16); and Hypoxia + S10 and Hypoxia + S30 groups (Hypoxia for 1st 4 weeks, hypoxia pretreated 10 mg/kg and 30 mg/kg salidroside by oral gavage per day for 2nd 4 weeks, n = 16 and 16). The excised hearts from four groups were measured by the heart weight index, H&E staining, TUNEL-positive assays and Western blotting.

Results

TUNEL-positive apoptotic cells in mice heart were less in Hypoxia + S10 and Hypoxia + S30 than those in the Hypoxia group. Compared with Hypoxia, the protein levels of Fas ligand, Fas death receptors, Fas-Associated Death Domain (FADD), activated caspase 8, and activated caspase 3 (Fas pathways) were decreased in Hypoxia + S10 and Hypoxia + S30. In the mitochondria pathway, the protein levels of BcLx, Bcl2, and Bid (anti-apoptotic Bcl2 family) in Hypoxia + S10 and Hypoxia + S30 were more than those in Hypoxia. The protein levels of Bax, t-Bid, activated caspase 9, and activated caspase 3 were less in Hypoxia + S10 and Hypoxia + S30 than those in hypoxia.

Conclusions

Our findings suggest that salidroside has protective effects on chronic intermittent hypoxia-induced Fas-dependent and mitochondria-dependent apoptotic pathways in mice hearts.

Introduction

Obstructive sleep apnea (OSA), a sleep breathing disorder, is associated with nocturnal airflow disruption in humans [1]. OSA is a high risk factor of cardiovascular diseases [2] and could increase the chance of heart failure by 140%, stroke by 60%, and coronary heart diseases by 30% [3], [4]. Chronic intermittent hypoxia (CIH) led to multiple long-term cardiovascular pathophysiologic consequences similar to what we observed in OSA [2], [5]. One study showed that CIH leads to left ventrical myocardial dysfunction [6] and our previous study showed that 8 week CIH induced cardiac abnormalities and apoptosis [7].

Salidroside [2-(4-hydroxyphenyl)ethyl beta-D-glucopyranoside], active ingredients of Rhodiola rosea, was used for high mountain sickness to protect erythrocytes against oxidative stress and improve resistance to stress and fatigue [8]. Salidroside was found to reduce cell apoptosis, improved cardiomyocyte glucose uptake, and reduced ischemia/reperfusion-induced cardiomyocyte damage [9]. However, the effect of salidroside on cardiovascular health is still not totally understood.

Apoptosis, a cell death program, has long been recognized to be involved in cardiovascular diseases [10], [11]. The cardiomyocyte apoptosis is one of the predictors of cardiac diseases or heart failure [12]. Cardiac widely dispersed apoptosis was found by our laboratory in chronic cardiometabolic or stressful conditions such as obesity [13], [14], [15], hypertension [11], [16], [17], diabetes [18], [19], ovariectomy [20], long-term hypoxia [7], [21], [22], and smoke [23]. The Fas receptor-dependent apoptotic (Type I) pathway is a major pathway triggering cardiac apoptosis [10], [24] and initiates binding the Fas ligand to the Fas receptor [24], [25], [26], [27]. This binding, followed by Fas-receptor oligomerization leading to the death-inducing signal complex, starts with recruitment of the Fas-Associated Death Domain (FADD) adaptor protein [24]. The activated caspase 8 cleaves pro-caspase 3 then undergoes autocatalysis to form active caspase 3, an effector caspase of apoptosis [24], [25]. The mitochondria-dependent (Type II) apoptotic pathway starts with apoptosis-regulating protein family exemplified by Bcl-2 family, such as anti-apoptotic Bcl-2 and pro-apoptotic Bad [24], [26], [27]. Pro-apoptotic Bcl2 family will enhance cytochrome c release from mitochondria [24], [26], [27], [28]. Cytochrome c release into cytosol activates caspase-9, then caspase-3 executes the apoptotic program [24], [27]. Besides, t-Bid was regarded as a main intracellular molecule signaling mediator from Fas to mitochondrial pathway because activated caspase 8 can cleave Bid to t-Bid then release cytochrome c to activate mitochondria-dependent apoptosis [24], [25]. In our previous study, the 8-week CIH was found to activate the Fas-dependent and mitochondria-dependent apoptotic pathways in rat hearts [7]. Salidroside protects H9c2 cells from ischemia/reperfusion-induced apoptosis through reduced cytochrome c release and caspase-3 activity [29] and attenuates apoptosis in ischemic hearts [30].

The effect of salidroside on CIH-induced cardiac apoptosis in mice hearts is not understood. We hypothesized that salidroside may prevent CIH-activated Fas-mediated and mitochondria-mediated cardiac apoptosis in mice hearts.

Section snippets

Animal model and salidroside

The studies were performed on sixty-four C57BL/6 J 5–6 month old male mice. Sixty-four C57BL/6 J mice 5–6 months of age were divided into four groups, the Control group (21% O₂, 24 h per day, 8 weeks, n = 16); Hypoxia group (Hypoxia: 7% O₂ 60 s, 20% O₂ alternating 60 s, 8 h per day, 8 weeks, n = 16); and Hypoxia + S10 group and Hypoxia + S30 group (Hypoxia for the first 4 weeks, Hypoxia pretreated 10 mg/kg and 30 mg/kg salidroside by oral gavage per day for the second 4 weeks, n = 16 and n = 16). Salidroside in the

Body weight and cardiac characteristics

Body weight (BW), whole heart weight normalized by body weight (WHW/BW), left ventricular weight normalized by body weight (LVW/BW), left ventricular weight normalized by whole heart weight (LVW/WHW), whole heart weight normalized by tibia length (WHW/Tibia), and left ventricular weight normalized by tibia length (LVW/Tibia) among the four groups, i.e. Control, Hypoxia, Hypoxia + S10 and Hypoxia + S30, were not different (Table 1). Interventricular septum at diastole (IVSd), left ventricular

Discussion

Our study identified the protective effects of salidroside treatment on heart disease associated with chronic intermittent hypoxia. The summarized findings are: (1) Cardiac fractional shortening was decreased after chronic intermittent hypoxia even though heart weight indices were of no difference. The treatment with salidroside showed improvement of cardiac fractional shortening subsequent to chronic intermittent hypoxia. (2) The activity of the cardiac Fas receptor-dependent apoptotic pathway

Acknowledgment

The study is supported by the National Science Council (NSC 98-2622-B-039-004-CC3; NSC 102-2410-H-468-029) and China Medical University (CMU99-COL-21), Taiwan. This study is also supported in part by the Taiwan Department of Health Clinical Trial and Research Center of Excellence (DOH102-TD-B-111-004). Research was supported by The Program for Professor of Special Appointment (Eastern Scholar, Honorary Chair Professor) at Shanghai Institute of Higher Education (No 2012-47).

References (38)

A. Bradford et al.
Does episodic hypoxia affect upper airway dilator muscle function? Implications for the pathophysiology of obstructive sleep apnoea
Respir Physiol Neurobiol
(2005)
S.D. Lee et al.
Effects of long-term intermittent hypoxia on mitochondrial and Fas death receptor dependent apoptotic pathways in rat hearts
Int J Cardiol
(2007)
T. Wu et al.
Cardioprotection of salidroside from ischemia/reperfusion injury by increasing N-acetylglucosamine linkage to cellular proteins
Eur J Pharmacol
(2009)
S.D. Lee et al.
Effects of exercise training on cardiac apoptosis in obese rats
Nutr Metab Cardiovasc Dis
(2013)
S.M. Cheng et al.
Exercise training enhances cardiac IGFI-R/PI3K/Akt and Bcl-2 family associated pro-survival pathways in streptozotocin-induced diabetic rats
Int J Cardiol
(2013)
S.D. Lee et al.
Cardiac Fas-dependent and mitochondria-dependent apoptosis in ovariectomized rats
Maturitas
(2008)
S.D. Lee et al.
Effects of short- and long-term hypobaric hypoxia on Bcl2 family in rat heart
Int J Cardiol
(2006)
N.H. Bishopric et al.
Molecular mechanisms of apoptosis in the cardiac myocyte
Curr Opin Pharmacol
(2001)
B.C. Barnhart et al.
The CD95 type I/type II model
Semin Immunol
(2003)
L. Sun et al.
Salidroside and tyrosol from Rhodiola protect H9c2 cells from ischemia/reperfusion-induced apoptosis
Life Sci
(2012)

H. Zhong et al.

Salidroside attenuates apoptosis in ischemic cardiomyocytes: a mechanism through a mitochondria-dependent pathway

J Pharmacol Sci

(2010)

K.Y. Zheng et al.

Salidroside stimulates the accumulation of HIF-1alpha protein resulted in the induction of EPO expression: a signaling via blocking the degradation pathway in kidney and liver cells

Eur J Pharmacol

(2012)

J. D'Armiento

Matrix metalloproteinase disruption of the extracellular matrix and cardiac dysfunction

Trends Cardiovasc Med

(2002)

L. Zhang et al.

Salidroside protects PC12 cells from MPP(+)-induced apoptosis via activation of the PI3K/Akt pathway

Food Chem Toxicol

(2012)

R.G.G. Tamisier et al.

A new model of chronic intermittent hypoxia in humans: effect on ventilation, sleep, and blood pressure

J Appl Physiol

(2009)

A.A. Chiang

Obstructive sleep apnea and chronic intermittent hypoxia: a review

Chin J Physiol

(2006)

E. Shahar et al.

Sleep-disordered breathing and cardiovascular disease: cross-sectional results of the Sleep Heart Health Study

Am J Respir Crit Care Med

(2001)

X. Yin et al.

Cardiac response to chronic intermittent hypoxia with a transition from adaptation to maladaptation: the role of hydrogen peroxide

Oxid Med Cell Longev

(2012)

L. Chen et al.

Oxidative stress and left ventricular function with chronic intermittent hypoxia in rats

Am J Respir Crit Care Med

(2005)

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Osteoarthritis (OA) was induced in rat knees by intra-articular injection of MIA; simultaneously salidroside was administered by intravenous injection. Pain behaviors were evaluated by knee-bend test, hind limb weight-bearing asymmetry and hind paw mechanical withdrawal threshold. The joint swelling was determined by the difference of knee joint diameter. Inflammatory exudates in synovial fluid were evaluated by leukocyte counting and protein content. Cytokines, chemokines, reactive oxygen species (ROS) and reactive nitrogen species (RNS) markers were determined by Enzyme-linked immunosorbent assay (ELISA) and colorimetric assay in synovial fluid. Pro-inflammatory gene expressions in synovial tissue were detected by quantitative real time RT-PCR (qRT-PCR). Nuclear factor kappa-B (NF-κB) DNA binding assay and western blot were used to determine NF-κB activation and ROS marker protein expression in synovial tissue. Glycosaminoglycan (GAG) content in the cartilage was measured by dimethylmethylene blue method. Hematoxylin and eosin (H&E), Safranin O-fast green and a modified Mankin grading system were used to evaluate the histology of articular cartilage.
Salidroside could alleviate pain and joint swelling in the early acute stage of OA in rat model, reduced the number of leukocytes, total protein content, proinflammatory mediators and ROS/RNS markers in synovial fluid, down regulated the expression of proinflammatory genes in synovium, inhibited the activation of NF- κ B and oxidative stress response in synovium, promoted the synthesis of cartilage GAG, prevented the loss of proteoglycan and chondrocyte degeneration.
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In China, Sal is widely used for the clinical prevention and treatment of acute mountain sickness and cardio-cerebrovascular diseases. Further studies have found that Sal could antagonize ischemic hypoxic cytotoxic effects by participating in several signaling pathways, including regulating HIF-1α, PI3K, MAPK, NF-κB signaling pathway [6–9], repressing caspase-dependent apoptosis, increasing EPO [9], VEGF [10], BcL-2 expression [3,11], and restoring the function of sodium and potassium channels [12]. However, the comprehensive protective mechanisms of Sal in mimic high aititude hypoxia remain obscure.
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¹: Chih-Yang Huang and Shin-Da Lee share equal contribution.

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Protective effect of salidroside on cardiac apoptosis in mice with chronic intermittent hypoxia

Abstract

Background

Methods

Results

Conclusions

Introduction

Section snippets

Animal model and salidroside

Body weight and cardiac characteristics

Discussion

Acknowledgment

Respir Physiol Neurobiol

Int J Cardiol

Eur J Pharmacol

Nutr Metab Cardiovasc Dis

Int J Cardiol

Maturitas

Int J Cardiol

Curr Opin Pharmacol

Semin Immunol

Life Sci

J Pharmacol Sci

Eur J Pharmacol

Trends Cardiovasc Med

Food Chem Toxicol

A new model of chronic intermittent hypoxia in humans: effect on ventilation, sleep, and blood pressure

J Appl Physiol

Obstructive sleep apnea and chronic intermittent hypoxia: a review

Chin J Physiol

Sleep-disordered breathing and cardiovascular disease: cross-sectional results of the Sleep Heart Health Study

Am J Respir Crit Care Med

Cardiac response to chronic intermittent hypoxia with a transition from adaptation to maladaptation: the role of hydrogen peroxide

Oxid Med Cell Longev

Oxidative stress and left ventricular function with chronic intermittent hypoxia in rats

Am J Respir Crit Care Med