Oxidative Medicine and Cellular Longevity

Oxidative Medicine and Cellular Longevity / 2017 / Article
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Antioxidant Phytochemicals at the Pharma-Nutrition Interface

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Research Article | Open Access

Volume 2017 |Article ID 3831750 | https://doi.org/10.1155/2017/3831750

Jung-Tung Liu, Huey-Yi Chen, Wen-Chi Chen, Kee-Ming Man, Yung-Hsiang Chen, "Red Yeast Rice Protects Circulating Bone Marrow-Derived Proangiogenic Cells against High-Glucose-Induced Senescence and Oxidative Stress: The Role of Heme Oxygenase-1", Oxidative Medicine and Cellular Longevity, vol. 2017, Article ID 3831750, 11 pages, 2017. https://doi.org/10.1155/2017/3831750

Red Yeast Rice Protects Circulating Bone Marrow-Derived Proangiogenic Cells against High-Glucose-Induced Senescence and Oxidative Stress: The Role of Heme Oxygenase-1

Academic Editor: Gian L. Russo
Received13 Jan 2017
Revised28 Mar 2017
Accepted02 Apr 2017
Published07 May 2017


The inflammation and oxidative stress of bone marrow-derived proangiogenic cells (PACs), also named endothelial progenitor cells, triggered by hyperglycemia contributes significantly to vascular dysfunction. There is supporting evidence that the consumption of red yeast rice (RYR; Monascus purpureus-fermented rice) reduces the vascular complications of diabetes; however, the underlying mechanism remains unclear. This study aimed to elucidate the effects of RYR extract in PACs, focusing particularly on the role of a potent antioxidative enzyme, heme oxygenase-1 (HO-1). We found that treatment with RYR extract induced nuclear factor erythroid-2-related factor nuclear translocation and HO-1 mRNA and protein levels in PACs. RYR extract inhibited high-glucose-induced (30 mM) PAC senescence and the development of reactive oxygen species (ROS) in a dose-dependent manner. The HO-1 inducer cobalt protoporphyrin IX also decreased high-glucose-induced cell senescence and oxidative stress, whereas the HO-1 enzyme inhibitor zinc protoporphyrin IX and HO-1 small interfering RNA significantly reversed RYR extract-caused inhibition of senescence and reduction of oxidative stress in high-glucose-treated PACs. These results suggest that RYR extract serves as alternative and complementary medicine in the treatment of these diseases, by inducing HO-1, thereby decreasing the vascular complications of diabetes.

1. Introduction

Endothelial dysfunction-related atherosclerosis is typically multifactorial. It is most often dependent on inflammatory risk factors such as hyperglycemia, hypercholesterolemia, hypertension, smoking, and obesity [1, 2]. Complications from atherosclerotic-related diseases remain the leading cause of mortality and morbidity in various industrialized countries [3].

Hyperglycemia, which is associated with endothelial dysfunction, is a primary cause of vascular complications in diabetes [4]. Evidence shows that the repair of endothelium involves bone marrow-derived proangiogenic cells (PACs), also known as endothelial progenitor cells (EPCs), in vasculogenesis [5]. The impaired function and reduced number of EPCs were found to be associated with vascular complications in both type I and type II diabetes [6, 7]. In addition, our previous studies have shown that hyperglycemia directly impairs the biological functions of angiogenesis, induces cellular aging (senescence), and produces reactive oxygen species (ROS) in EPCs [811].

The definition of EPCs has changed over the years as many studies have revealed the true face of the majority of EPC heterogeneity, which are in fact not endothelial precursors but can be described as myeloid-lineage-derived cells with proangiogenic properties. EPCs were classically described as cells that expressed a combination of endothelial and progenitor markers; however, none of these markers is fully specific [1214]. Thus, other names, such as bone-marrow-derived PACs, have been suggested for EPCs. Nevertheless, despite their history and controversy, EPCs have been applied to different cell types that play roles in the regeneration of the endothelial lining in vasculature. EPCs in all their forms remain a promising target of regenerative medicine.

Red yeast rice (RYR; Monascus purpureus Went-fermented rice) has been used for many centuries to make rice wine in China, to maintain food taste and color, and for its medicinal properties. Biological and epidemiological evidence support that the intake of RYR may reduce the incidence of atherosclerosis. RYR contains naturally occurring statins that have serum lipid-modulating effects. Thus, RYR has a lipid-lowering effect in subjects with hyperlipidemia. Pharmacological RYR-related products are marketed in China, Taiwan, and in the United States. RYR has also been shown to have free radical scavenging abilities and can protect the function of endothelium through antioxidative and anti-inflammatory mechanisms [15]. Moreover, a previous study showed that RYR inhibited homocysteine-induced endothelial adhesion via intracellular ROS reduction [16].

Heme oxygenase-1 (HO-1) is a member of the heat shock protein family. The expression of HO-1 is triggered by various stressors, including oxidative stress, heavy metals, UV radiation, and hypoxia [17]. HO-1 expression is mediated through accumulation of the nuclear factor erythroid-2-related factor (Nrf2) in the nucleus [18]. HO-1 was found to be a pivotal antioxidative, anti-inflammatory, and antiapoptotic molecule [19]. Various medicinal plant-derived chemical substances may induce HO-1 activation and can maximize the intrinsic antioxidative abilities [20].

In this study, we explored the potency of RYR extract as an HO-1 inducer in PACs and investigated whether it contributed to the beneficial effects against PAC senescence and oxidative stress.

2. Materials and Methods

2.1. Materials

Glucose, mannitol, and other chemicals were obtained from Sigma Chemical Co. (MO, USA). RYR (LipoCol Forte) was obtained from NatureWise Biotech & Medicals Corporation (Taipei, Taiwan) and extracted at room temperature [15, 16]. Final concentration of solvents in following studies was always less than 0.5% to avoid potential interference. The cobalt protoporphyrin IX (CoPPIX) and zinc protoporphyrin IX (ZnPPIX) used (10 μM) did not significantly influence cell viability (>90%).

2.2. PAC Isolation, Cultivation, and Identification

The protocol conforms to the Helsinki declaration. China Medical University (Taichung, Taiwan) Institutional Review Board approved the study by expedited review. Peripheral blood mononuclear cells (MNCs) were isolated (gradient centrifugation) from volunteers by Histopaq-1077 (Sigma, USA). Isolated MNCs were plated in endothelial growth medium (EGM-2 MV; Cambrex, USA), with supplements (hydrocortisone, R3-insulin-like growth factor 1, human epidermal growth factor, VEGF, human fibroblast growth factor, gentamicin, amphotericin B, vitamin C, and 20% fetal bovine serum) on fibronectin-coated plates. After culturing, medium was replaced and nonadherent cells were removed. Culture medium was changed every 3 days, and a number of cells can continue to grow into late outgrowth cells. Late outgrowth PACs under passage 3 were used for the study [8].

PACs were further characterized by immunofluorescence for CD34, kinase insert domain receptor (KDR, also named vascular endothelial growth factor receptor 2), and CD31 (platelet endothelial cell adhesion molecule; PECAM-1) (Santa Cruz, USA) expressions [8].

2.3. Western Blot Analysis

Cell lysates were prepared in lysis buffer (20 mM Tris-HCl, 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 1% Triton, 2.5 mM sodium pyrophosphate, 1 mM glycerophosphate, 1 mM Na3VO4, 1 μg/ml leupeptin, and 1 mM phenylmethylsulfonyl fluoride, pH 7.5). The concentration of protein was determined by the Bio-Rad Protein Assay reagent.

Nuclear protein extracts were prepared as previously described [8]. In brief, after being washed with ice-cold PBS, cells were scraped off the plates with a cell scraper in 1 mL of ice-cold buffer A (10 mmol/l HEPES/NaOH, pH 7.9; 10 mmol/l KCl; 1.5 mmol/l MgCl2; 1 mmol/l DTT; l0.5 mmol/l PMSF; 2 μg/ml aprotinin; 2 μg/ml pepstatin; and 2 μg/ml leupeptin). After centrifugation at 300 for 10 minutes at 4°C, cells were resuspended in 80 μl of buffer B (buffer A containing 0.1% Triton X-100) by gentle pipetting. Cell lysates were allowed to stand on ice for 10 minutes and then centrifuged at 12,000 for 10 minutes at 4°C. Nuclear pellets were resuspended in 70 μL of ice-cold buffer C (20 mmol/l HEPES/NaOH, pH 7.9; 1.5 mmol/ MgCl2; 1 mmol/l DTT; 0.2 mmol/l EDTA; 420 mmol/l NaCl; 25% glycerol; 0.5 mmol/l PMSF; 2 μg/ml aprotinin; 2 μg/ml pepstatin; and 2 μg/ml leupeptin), incubated on ice for 30 minutes with intermittent mixing, and then centrifuged at 15,000 for 30 minutes at 4°C. Nuclear protein extracts prepared as described above were determined by protein assay.

Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membrane. The membranes were probed with goat anti-HO-1 antibody (R&D Systems, MN, USA) or rabbit anti-Nrf2 antibody (Abcam, Cambridge, MA, USA) and then incubated with horseradish peroxidase-conjugated secondary antibodies, and the proteins were visualized with a chemiluminescence detection kit (Amersham Biosciences, NJ, USA). Mouse anti-β-actin (Labvision/NeoMarkers, CA, USA) or anti-lamin B (Abcam, Cambridge, MA, USA) antibodies were used as loading controls. Protein expression levels were quantified using ImageQuant (USA) software [20].

2.4. PAC Viability

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT, Sigma, USA) assay was used for cell viability. Briefly, PACs were grown in plates and incubated with various concentrations of agents. Medium containing MTT (0.5 mg/ml) was added. Finally, dimethyl sulfoxide was added to each well and the absorbance of blue formazan read at 540 nm using a microplate reader (Multiskan Ex, Thermo Lab systems, USA). Cells incubated in control medium were considered 100% viable [8].

2.5. PAC Senescence Assay

The senescence of PACs was determined by the senescent cell staining kit (Sigma, USA). Briefly, PACs were fixed for 6 min in 2% formaldehyde and 0.2% glutaraldehyde in phosphate-buffered saline and then incubated for 12 h at 37°C with fresh X-gal staining solution (1 mg/ml X-gal, 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, and 2 mM MgCl2; pH 6). Then, blue-stained and total cells were counted for calculating the β-galactosidase-positive cell percentage [8].

2.6. ROS Production

Effect of RYR extract on ROS production was determined by a fluorometric assay by probe dichloro-dihydro-fluorescein diacetate (DCFH-DA). Confluent cells in 48-well plates were pretreated with RYR extract. HBSS containing 10 μM DCFH-DA was added, and the cells were incubated with it for 30 min. The relative fluorescence unit was measured at 485 nm excitation and 530 nm emission by a fluorescence microplate reader [21].

2.7. Measurement of Glutathione (GSH), Glutathione Reductase (GR), and Thiobarbituric Acid Reactive Substances (TBARS)

GSH levels were measured by a colorimetric assay (Bioxytech GSH-400; OxisResearch, Portland, OR, USA). Metaphosphoric acid (5%) was added to the cells and then scraped off it. The mixture was centrifuged at 3000 for 5 min at 4°C, and the supernatant was measured at 400 nm after a chemical reaction with reagent R1 (4-chloro-1-methyl-7-trifluromethyl-quinolinium methylsulfate) and reagent R2 (30% NaOH) with a GSH standard curve [22].

The GR activity was determined with a protocol described previously [23]. In brief, GR activity was expressed as a rate of decrease in absorbance at 340 nm/min due to the NADPH oxidation by GR, and the enzyme activity was normalized with mg protein.

Lipid peroxidation was quantified by TBARS determination by spectrophotometric assay (Beckman Coulter, DU 640 spectrophotometer, Germany). The lipid peroxide levels, expressed as nmol malondialdehyde/mg protein, were calculated from the absorbance at 532 nm by external standard tetraethoxypropane [22].

2.8. RNA Extraction and Real-Time PCR

Total RNA was isolated from lung cancer tissues and adjacent normal lung tissues of the NSCLC patients and, subsequently, analyzed by real-time PCR. The following primers were designed using Primer Express software (RealQuant, Roche) based on published sequences: human HO-1 sense primer: 5′-TTC TTC ACC TTC CCC AAC TA-3′; HO-1 antisense primer 5′-GCA TAA AGC CCT ACA GCA AC-3′. Human GAPDH sense primer: 5′-AGC CAC ATC GCT CAG ACA-3′; GAPDH antisense primer 5′-GCC CAA TAC GAC CAA ATC C-3′. Fluorescence data were acquired after the final extension step. A melt analysis was conducted for all products to determine the specificity of the amplification [24].

2.9. Small Interfering RNA

A specific double-stranded 21-nucleotide RNA sequence homologous (small interfering RNA (siRNA)) to the target gene was used to silence HO-1 expression. The computer software and Silencer™ siRNA construction kit from Ambion (Austin, TX, USA) designed and synthesized siRNA for HO-1 (sequences of the ribonucleotides were 5′-rGAC UGC GUU CCU GCU CAA CdTdT-3′ and 5′-rGUU GAG CAG GAA CGC AGU CdTdT-3′) and negative control number 1 siRNA. HO-1 protein inhibition was assessed by immunoblot analysis following transfection of cells with HO-1-siRNA. Briefly, cells were transiently transfected with 20 nM siRNA using 8 μl of siPORT Amine (Ambion) [24, 25].

2.10. Statistical Analyses

Data were expressed as means ± standard deviation (SD). Statistical evaluation was performed using Student’s t-test or one-way analysis of variance, followed by Dunnett’s test. A value of <0.05 was considered significant.

3. Results

3.1. Isolation and Characterization of Circulating Bone Marrow-Derived PACs

Cells originated from peripheral blood MNCs of healthy volunteers. MNCs initially seeded on fibronectin-coated wells were round-shaped. Late outgrowth PACs with cobblestone-like morphology were grown to confluence (Figure 1).

Cell characterization was performed by fluorescent stain. CD34, KDR, and CD31 may be considered markers of late outgrowth PACs. CD34 and KDR double positive may be important markers of these cells in vitro (Figure 1) [8, 26]. Endothelial marker CD31 was also used for characterization of the outgrowth cells.

3.2. RYR Extract Is Toxic Only in High Concentrations

Incubation of PACs with 0–50 μg/ml RYR extract for 24 h and 48 h did not result in cellular toxicity; however, high doses of RYR extract (≥200 μg/ml for 24 h and ≥100 μg/ml for 48 h) significantly reduced cell viability (Figure 2). These data indicate that the significant cytotoxic effects of RYR extract on PACs were found in high doses. Thus, the noncytotoxic doses of RYR extract (≤50 μg/ml) in the following experiments were used to avoid potential interference of cell survival.

3.3. RYR Extract Induces Nrf-2 Activation and HO-1 Expression in PACs

We further tested the effects of RYR extract on Nrf2 signaling pathway and HO-1 expression in PACs. 50 μg/ml RYR extract time-dependently induced Nrf2 nuclear translocation in PAC cells (Figure 3(a)). In addition, RYR extract (12.5, 25, and 50 μg/ml) was added to culture medium with PACs, and real-time PCR and Western blot were performed for HO-1 mRNA (12 h) and protein (48 h) expressions, respectively. As shown in Figures 3(b) and 3(c), RYR extract increased HO-1 mRNA and protein expression in a dose-dependent manner. Moreover, 50 μg/ml of RYR extract increased HO-1 protein expression in PACs in a time-dependent manner (12, 24, and 48 h) (Figure 3(d)).

3.4. RYR Extract Inhibits High-Glucose-Induced Senescence and Oxidative Stress

Our previous study demonstrated that high-glucose-caused (30 mM) senescence and oxidative stress in PACs as compared with the control (5 mM of glucose) or osmotic control (extra 25 mM of mannitol) groups [8]. To investigate whether RYR extract inhibited senescence of PACs induced by high glucose, PACs were coincubated with high glucose (30 mM) and RYR extract (12.5–50 μg/ml) for 48 h, and a β-galactosidase assay was performed. RYR extract had a dose-dependent effect to reduce senescence in high-glucose-treated PACs (Figure 4(a)).

In addition, to directly determine the effect of RYR extract on ROS generation, we analyzed the ROS level in high-glucose-treated PACs. As shown in Figure 4(b), treatment with high glucose for 48 h caused a higher increase of fluorescence compared with the control and mannitol groups. Coincubation of PACs with RYR extract inhibited high-glucose-induced ROS generation in a dose-dependent manner.

In addition, Table 1 shows that RYR extract treatment caused a significant increase of GSH content and GR activity but a significant decrease of thiobarbituric acid reactive substance (TBARS) content relative to high-glucose-treated PACs.

ControlMannitolGlucoseGlucose + RYR extract

GSH (nmol/mg protein)52.6 ± 8.346.3 ± 3.929.3 ± 2.648.5 ± 5.5#
GR (unit/mg protein)1.8 ± 0.21.6 ± 0.21.1 ± 0.11.6 ± 0.1#
TBARS (nmol/mg protein)2.8 ± 0.53.2 ± 0.68.2 ± 1.13.6 ± 1.9#

compared to that of the control group; compared to that of the glucose group.
3.5. RYR Extract Inhibits High-Glucose-Induced Senescence and Oxidative Stress via HO-1

HO-1 siRNA was used to confirm the effects of RYR extract mediated through HO-1. Figure 5 shows a reduction of RYR extract-induced HO-1 by HO-1 siRNA. We further explored the effect of HO-1 on high-glucose-induced PAC senescence and oxidative stress. As shown in Figures 6(a) and 6(b), we found that the HO-1 inducer cobalt protoporphyrin (CoPPIX) also significantly decreased high-glucose-induced PAC senescence and oxidative stress, whereas the HO-1 enzyme inhibitor, zinc protoporphyrin IX (ZnPPIX), and HO-1 siRNA significantly reversed RYR extract-caused inhibition in high-glucose-treated PACs. Oxidative stimulator H2O2 (50 μM) was also used to confirm the antioxidative effect of RYR extract.

4. Discussion

The present study showed, for the first time, that RYR extract attenuated high-glucose-induced senescence and oxidative stress of PACs. Our data also suggest that HO-1 activation may play a pivotal role in the anticellular senescence and antioxidative effects of RYR extract on PACs.

As the incidence of myocardial infarction and stroke increases as the population ages, there must be an increased focus on the fundamental processes and mechanisms of vascular aging. An advanced understanding of the molecular pathways leading to vascular aging may contribute to the design of therapeutic strategies to prevent vascular senescence. More recently, Paneni et al. reviewed the advances in the pathology of age-related vascular dysfunction including dysregulation of epigenetic modifications, inflammatory genes, and mechanisms of vascular calcification [27]. Oxidative stress contributes to the progression of endothelial-dysfunction-related clinical diseases through luminal narrowing in the brain (ischemic stroke), heart, and peripheral vessels [28]. Epidemiological studies indicate that RYR consumption is associated with reduced coronary heart disease risk [29, 30]. In vitro investigations have indicated that RYR can inhibit several key events of the atherogenic process, such as vascular smooth muscle and endothelial cell dysfunction by redox-sensitive mechanisms [15, 16, 31]. RYR contains chemicals that are similar to prescription statin medications including monacolin K (the same structure as the drug lovastatin). Statin-mediated HO-1 induction has been shown to occur in vascular smooth muscle cells [32, 33], endothelial cells [3440], macrophages [41, 42], neurons [4346], liver cells [47, 48], and pulmonary cells [49, 50]. In this study, we showed, for the first time, that RYR extract induces HO-1 activation in bone-marrow-derived PACs. HO-1 expression is mediated through accumulation of Nrf2 in the nucleus. We also found that RYR extract induces Nrf2 nuclear translocation in PACs. Moreover, the antioxidant properties of RYR extract were further reported to protect against cellular senescence by inhibiting high-glucose-induced oxidative stress in PACs via HO-1 induction. Thus, our results elucidated the relationship with hyperglycemia, oxidative stress, and endothelial dysfunction, regulation of atheroprotective genes HO-1, and how the regulation of these activities by RYR can lead to the prevention of diabetes-related vascular complications.

During atherogenesis, hyperglycemia-mediated chronic oxidative stress plays an important role in PAC dysfunction [51]. In the present study, RYR extract induced HO-1 expression in PACs in a dose- and time-dependent manner. Consistent with the present results, various vascular protective agents such as atorvastatin [52], estradiol [53], and oleuropein/oleacein (phenolic compounds from olive oil) [54] also increase HO-1 and display anti-inflammatory effects in EPCs. All these data suggest that RYR is a strong inducer of HO-1, and such induction may be independent of various vascular protective agents.

ROS have been implicated in the pathogenesis of most stages of atherosclerosis [11, 55, 56]. ROS, especially hydrogen peroxide and superoxide, are important intracellular signaling molecules in cells. ROS participate in the growth and death of PACs; these events play crucial roles in cardiovascular diseases, suggesting that the sources of ROS and the intracellular signaling pathways may be important therapeutic targets [57]. Evidence has shown that ROS influences cellular processes in vascular remodeling by activating various intracellular signaling cascades [57]. Our previous in vitro study demonstrated the activity of RYR extract on the radical-scavenging abilities of the probe-based ultraweak chemiluminescence technique and showed that RYR exhibited major radical-scavenging abilities on superoxide and hydroxyl radicals [16]. The present study further provides direct evidence that RYR extract maintained GSH amounts and upregulated GR activity in high-glucose-stressed PACs resulting in decreased TBARS, suggesting that RYR could maintain the intracellular antioxidant concentrations in biological systems. It should be further examined whether RYR upregulates other GSH-related enzymes, such as glutamate cysteine ligase and glutathione peroxidase, which catalyze GSH biosynthesis.

Our study has limitations. The composition of the various compounds from the RYR extract, in particular those that might be responsible for the protective effects in the RYR mixture and may specifically induce HO-1, is not clearly defined in this manuscript; only the crude extract of RYR was studied. Moreover, the molecular mechanism underlying HO-1 induction by RYR is unclear. For example, it is unknown if nuclear factor erythroid 2-related factor 2, a major transcriptional regulator of HO-1 expression [58], or the HO products carbon monoxide and bilirubin [59] were involved in the protective effects of RYR on PACs. Finally, the measurement of some direct inflammation markers could provide more evidence on the anti-inflammatory role of HO-1 after RYR treatment. It is important to explore the effective compounds and mechanisms of RYR for further direction in the field of agricultural product research.

5. Conclusions

The present study demonstrated that RYR extract induced HO-1 expression in PACs in a dose- and time-dependent manner. RYR extract inhibited high-glucose-induced β-galactosidase activation and reduced high-glucose-induced oxidative stress in PACs in a dose-dependent manner. HO-1 expression might play a pivotal role in the atheroprotective effects of RYR on PACs. Thus, RYR may fulfill the definition of a pharmacological preconditioning agent for preventing cerebrovascular and cardiovascular diseases.

Conflicts of Interest

The authors declare that they have no conflicts of interest.


This work was supported in part by China Medical University (CMU104-S-37), Chung Shan Medical University Hospital (CS08110), Taiwan Ministry of Science and Technology (MOST 104-2320-B-039-016-MY3), Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW106-TDU-B-212-113004), and CMU under the Aim for Top University Plan of the Ministry of Education, Taiwan.


  1. E. Profumo, B. Buttari, D. D'Arcangelo et al., “The nutraceutical dehydrozingerone and its dimer counteract inflammation- and oxidative stress-induced dysfunction of in vitro cultured human endothelial cells: a novel perspective for the prevention and therapy of atherosclerosis,” Oxidative Medicine and Cellular Longevity, vol. 2016, Article ID 1246485, p. 12, 2016. View at: Publisher Site | Google Scholar
  2. L. Xiao, J. H. Dong, S. Jin et al., “Hydrogen sulfide improves endothelial dysfunction via downregulating BMP4/COX-2 pathway in rats with hypertension,” Oxidative Medicine and Cellular Longevity, vol. 2016, Article ID 8128957, p. 10, 2016. View at: Publisher Site | Google Scholar
  3. K. E. Watson, E. H. Stocker, D. S. Jacoby, and P. A. McCullough, “Advanced lipid testing: when, why, and in whom?” Reviews in Cardiovascular Medicine, vol. 15, no. 4, pp. 310–317, 2014. View at: Google Scholar
  4. C. L. Jao, C. C. Hung, Y. S. Tung, P. Y. Lin, M. C. Chen, and K. C. Hsu, “The development of bioactive peptides from dietary proteins as a dipeptidyl peptidase IV inhibitor for the management of type 2 diabetes,” Biomedicine (Taipei), vol. 5, no. 3, p. 14, 2015. View at: Publisher Site | Google Scholar
  5. K. Y. Howangyin and J. S. Silvestre, “Diabetes mellitus and ischemic diseases: molecular mechanisms of vascular repair dysfunction,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 34, no. 6, pp. 1126–1135, 2014. View at: Publisher Site | Google Scholar
  6. N. Lois, R. V. McCarter, C. O'Neill, R. J. Medina, and A. W. Stitt, “Endothelial progenitor cells in diabetic retinopathy,” Frontiers in Endocrinology, vol. 5, p. 44, 2014. View at: Publisher Site | Google Scholar
  7. K. H. Yiu and H. F. Tse, “Specific role of impaired glucose metabolism and diabetes mellitus in endothelial progenitor cell characteristics and function,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 34, no. 6, pp. 1136–1143, 2014. View at: Publisher Site | Google Scholar
  8. Y. H. Chen, S. J. Lin, F. Y. Lin et al., “High glucose impairs early and late endothelial progenitor cells by modifying nitric oxide-related but not oxidative stress-mediated mechanisms,” Diabetes, vol. 56, no. 6, pp. 1559–1568, 2007. View at: Publisher Site | Google Scholar
  9. P. H. Huang, H. Y. Tsai, C. H. Wang et al., “Moderate intake of red wine improves ischemia-induced neovascularization in diabetic mice—roles of endothelial progenitor cells and nitric oxide,” Atherosclerosis, vol. 212, no. 2, pp. 426–435, 2010. View at: Publisher Site | Google Scholar
  10. P. H. Huang, J. S. Chen, H. Y. Tsai et al., “Globular adiponectin improves high glucose-suppressed endothelial progenitor cell function through endothelial nitric oxide synthase dependent mechanisms,” Journal of Molecular and Cellular Cardiology, vol. 51, no. 1, pp. 109–119, 2011. View at: Publisher Site | Google Scholar
  11. C. P. Lin, F. Y. Lin, P. H. Huang et al., “Endothelial progenitor cell dysfunction in cardiovascular diseases: role of reactive oxygen species and inflammation,” BioMed Research International, vol. 2013, Article ID 845037, p. 10, 2013. View at: Publisher Site | Google Scholar
  12. C. L. Barber and M. L. Iruela-Arispe, “The ever-elusive endothelial progenitor cell: identities, functions and clinical implications,” Pediatric Research, vol. 59, Part 2, no. 4, pp. 26R–32R, 2006. View at: Publisher Site | Google Scholar
  13. J. Case, L. E. Mead, W. K. Bessler et al., “Human CD34+AC133+VEGFR-2+ cells are not endothelial progenitor cells but distinct, primitive hematopoietic progenitors,” Experimental Hematology, vol. 35, no. 7, pp. 1109–1118, 2007. View at: Publisher Site | Google Scholar
  14. F. Timmermans, J. Plum, M. C. Yoder, D. A. Ingram, B. Vandekerckhove, and J. Case, “Endothelial progenitor cells: identity defined?” Journal of Cellular and Molecular Medicine, vol. 13, no. 1, pp. 87–102, 2009. View at: Publisher Site | Google Scholar
  15. C. P. Lin, P. H. Huang, H. S. Tsai et al., “Monascus purpureus-fermented rice inhibits tumor necrosis factor-alpha-induced upregulation of matrix metalloproteinase 2 and 9 in human aortic smooth muscle cells,” Journal of Pharmacy and Pharmacology, vol. 63, no. 12, pp. 1587–1594, 2011. View at: Publisher Site | Google Scholar
  16. C. P. Lin, Y. H. Chen, J. W. Chen et al., “Cholestin (Monascus purpureus rice) inhibits homocysteine-induced reactive oxygen species generation, nuclear factor-kappaB activation, and vascular cell adhesion molecule-1 expression in human aortic endothelial cells,” Journal of Biomedical Science, vol. 15, no. 2, pp. 183–196, 2008. View at: Publisher Site | Google Scholar
  17. M. D. Hjortso and M. H. Andersen, “The expression, function and targeting of haem oxygenase-1 in cancer,” Current Cancer Drug Targets, vol. 14, no. 4, pp. 337–347, 2014. View at: Publisher Site | Google Scholar
  18. N. Li, J. Alam, M. I. Venkatesan et al., “Nrf2 is a key transcription factor that regulates antioxidant defense in macrophages and epithelial cells: protecting against the proinflammatory and oxidizing effects of diesel exhaust chemicals,” Journal of Immunology, vol. 173, no. 5, pp. 3467–3481, 2004. View at: Publisher Site | Google Scholar
  19. Y. M. Kim, H. O. Pae, J. E. Park et al., “Heme oxygenase in the regulation of vascular biology: from molecular mechanisms to therapeutic opportunities,” Antioxidants & Redox Signaling, vol. 14, no. 1, pp. 137–167, 2011. View at: Publisher Site | Google Scholar
  20. P. L. Liu, J. T. Liu, H. F. Kuo, I. W. Chong, and C. C. Hsieh, “Epigallocatechin gallate attenuates proliferation and oxidative stress in human vascular smooth muscle cells induced by interleukin-1beta via heme oxygenase-1,” Mediators of Inflammation, vol. 2014, Article ID 523684, p. 8, 2014. View at: Publisher Site | Google Scholar
  21. J. W. Chen, Y. H. Chen, F. Y. Lin, Y. L. Chen, and S. J. Lin, “Ginkgo biloba extract inhibits tumor necrosis factor-alpha-induced reactive oxygen species generation, transcription factor activation, and cell adhesion molecule expression in human aortic endothelial cells,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 23, no. 9, pp. 1559–1566, 2003. View at: Publisher Site | Google Scholar
  22. Y. H. Chen, F. Y. Lin, P. L. Liu et al., “Antioxidative and hepatoprotective effects of magnolol on acetaminophen-induced liver damage in rats,” Archives of Pharmacal Research, vol. 32, no. 2, pp. 221–228, 2009. View at: Publisher Site | Google Scholar
  23. S. K. Jain and D. Micinski, “Vitamin D upregulates glutamate cysteine ligase and glutathione reductase, and GSH formation, and decreases ROS and MCP-1 and IL-8 secretion in high-glucose exposed U937 monocytes,” Biochemical and Biophysical Research Communications, vol. 437, no. 1, pp. 7–11, 2013. View at: Publisher Site | Google Scholar
  24. J. R. Tsai, H. M. Wang, P. L. Liu et al., “High expression of heme oxygenase-1 is associated with tumor invasiveness and poor clinical outcome in non-small cell lung cancer patients,” Cellular Oncology (Dordrecht), vol. 35, no. 6, pp. 461–471, 2012. View at: Publisher Site | Google Scholar
  25. P. L. Liu, J. R. Tsai, A. L. Charles et al., “Resveratrol inhibits human lung adenocarcinoma cell metastasis by suppressing heme oxygenase 1-mediated nuclear factor-kappaB pathway and subsequently downregulating expression of matrix metalloproteinases,” Molecular Nutrition & Food Research, vol. 54, Supplement 2, pp. S196–S204, 2010. View at: Publisher Site | Google Scholar
  26. C. H. Yoon, J. Hur, K. W. Park et al., “Synergistic neovascularization by mixed transplantation of early endothelial progenitor cells and late outgrowth endothelial cells: the role of angiogenic cytokines and matrix metalloproteinases,” Circulation, vol. 112, no. 11, pp. 1618–1627, 2005. View at: Publisher Site | Google Scholar
  27. F. Paneni, S. Costantino, and F. Cosentino, “Molecular pathways of arterial aging,” Clinical Science (London, England: 1979), vol. 128, no. 2, pp. 69–79, 2015. View at: Publisher Site | Google Scholar
  28. P. Pantcheva, M. Elias, K. Duncan, C. V. Borlongan, N. Tajiri, and Y. Kaneko, “The role of DJ-1 in the oxidative stress cell death cascade after stroke,” Neural Regeneration Research, vol. 9, no. 15, pp. 1430–1433, 2014. View at: Publisher Site | Google Scholar
  29. Y. Feng, H. Xu, and K. Chen, “Natural polypill Xuezhikang: its clinical benefit and potential multicomponent synergistic mechanisms of action in cardiovascular disease and other chronic conditions,” Journal of Alternative and Complementary Medicine, vol. 18, no. 4, pp. 318–328, 2012. View at: Publisher Site | Google Scholar
  30. C. W. Yang and S. A. Mousa, “The effect of red yeast rice (Monascus purpureus) in dyslipidemia and other disorders,” Complementary Therapies in Medicine, vol. 20, no. 6, pp. 466–474, 2012. View at: Publisher Site | Google Scholar
  31. C. P. Lin, Y. L. Lin, P. H. Huang, H. S. Tsai, and Y. H. Chen, “Inhibition of endothelial adhesion molecule expression by Monascus purpureus-fermented rice metabolites, monacolin K, ankaflavin, and monascin,” Journal of the Science of Food and Agriculture, vol. 91, no. 10, pp. 1751–1758, 2011. View at: Publisher Site | Google Scholar
  32. T. S. Lee, C. C. Chang, Y. Zhu, and J. Y. Shyy, “Simvastatin induces heme oxygenase-1: a novel mechanism of vessel protection,” Circulation, vol. 110, no. 10, pp. 1296–1302, 2004. View at: Publisher Site | Google Scholar
  33. M. Li, Y. Liu, H. Shi et al., “Statins inhibit pulmonary artery smooth muscle cell proliferation by upregulation of HO-1 and p21WAF1,” Naunyn-Schmiedebergs Archives of Pharmacology, vol. 385, no. 10, pp. 961–968, 2012. View at: Publisher Site | Google Scholar
  34. N. Grosser, K. Erdmann, A. Hemmerle et al., “Rosuvastatin upregulates the antioxidant defense protein heme oxygenase-1,” Biochemical and Biophysical Research Communications, vol. 325, no. 3, pp. 871–876, 2004. View at: Publisher Site | Google Scholar
  35. N. Grosser, A. Hemmerle, G. Berndt et al., “The antioxidant defense protein heme oxygenase 1 is a novel target for statins in endothelial cells,” Free Radical Biology and Medicine, vol. 37, no. 12, pp. 2064–2071, 2004. View at: Publisher Site | Google Scholar
  36. J. Dulak, A. Loboda, A. Jazwa et al., “Atorvastatin affects several angiogenic mediators in human endothelial cells,” Endothelium, vol. 12, no. 5-6, pp. 233–241, 2005. View at: Publisher Site | Google Scholar
  37. F. Ali, S. S. Hamdulay, A. R. Kinderlerer et al., “Statin-mediated cytoprotection of human vascular endothelial cells: a role for Kruppel-like factor 2-dependent induction of heme oxygenase-1,” Journal of Thrombosis and Haemostasis, vol. 5, no. 12, pp. 2537–2546, 2007. View at: Publisher Site | Google Scholar
  38. F. Ali, M. Zakkar, K. Karu et al., “Induction of the cytoprotective enzyme heme oxygenase-1 by statins is enhanced in vascular endothelium exposed to laminar shear stress and impaired by disturbed flow,” Journal of Biological Chemistry, vol. 284, no. 28, pp. 18882–18892, 2009. View at: Publisher Site | Google Scholar
  39. U. Hinkelmann, N. Grosser, K. Erdmann, H. Schroder, and S. Immenschuh, “Simvastatin-dependent up-regulation of heme oxygenase-1 via mRNA stabilization in human endothelial cells,” European Journal of Pharmaceutical Sciences, vol. 41, no. 1, pp. 118–124, 2010. View at: Publisher Site | Google Scholar
  40. K. J. Kim, K. S. Kim, N. R. Kim, and H. S. Chin, “Effects of simvastatin on the expression of heme oxygenase-1 in human RPE cells,” Investigative Ophthalmology and Visual Science, vol. 53, no. 10, pp. 6456–6464, 2012. View at: Publisher Site | Google Scholar
  41. J. C. Chen, K. C. Huang, and W. W. Lin, “HMG-CoA reductase inhibitors upregulate heme oxygenase-1 expression in murine RAW264.7 macrophages via ERK, p38 MAPK and protein kinase G pathways,” Cellular Signalling, vol. 18, no. 1, pp. 32–39, 2006. View at: Publisher Site | Google Scholar
  42. P. O. Leung, S. H. Wang, S. H. Lu, W. H. Chou, C. Y. Shiau, and T. C. Chou, “Simvastatin inhibits pro-inflammatory mediators through induction of heme oxygenase-1 expression in lipopolysaccharide-stimulated RAW264.7 macrophages,” Toxicology Letters, vol. 207, no. 2, pp. 159–166, 2011. View at: Publisher Site | Google Scholar
  43. C. H. Hsieh, C. S. Rau, M. W. Hsieh et al., “Simvastatin-induced heme oxygenase-1 increases apoptosis of neuro 2A cells in response to glucose deprivation,” Toxicological Sciences, vol. 101, no. 1, pp. 112–121, 2008. View at: Publisher Site | Google Scholar
  44. C. H. Hsieh, S. F. Jeng, M. W. Hsieh et al., “Statin-induced heme oxygenase-1 increases NF-kappaB activation and oxygen radical production in cultured neuronal cells exposed to lipopolysaccharide,” Toxicological Sciences, vol. 102, no. 1, pp. 150–159, 2008. View at: Publisher Site | Google Scholar
  45. C. H. Hsieh, J. C. Jeng, M. W. Hsieh et al., “Involvement of the p38 pathway in the differential induction of heme oxygenase-1 by statins in neuro-2A cells exposed to lipopolysaccharide,” Drug and Chemical Toxicology, vol. 34, no. 1, pp. 8–19, 2011. View at: Publisher Site | Google Scholar
  46. D. A. Butterfield, E. Barone, F. Di Domenico et al., “Atorvastatin treatment in a dog preclinical model of Alzheimer’s disease leads to up-regulation of haem oxygenase-1 and is associated with reduced oxidative stress in brain,” International Journal of Neuropsychopharmacology, vol. 15, no. 7, pp. 981–987, 2012. View at: Publisher Site | Google Scholar
  47. I. R. Lai, K. J. Chang, H. W. Tsai, and C. F. Chen, “Pharmacological preconditioning with simvastatin protects liver from ischemia-reperfusion injury by heme oxygenase-1 induction,” Transplantation, vol. 85, no. 5, pp. 732–738, 2008. View at: Publisher Site | Google Scholar
  48. I. G. Habeos, P. G. Ziros, D. Chartoumpekis, A. Psyrogiannis, V. Kyriazopoulou, and A. G. Papavassiliou, “Simvastatin activates Keap1/Nrf2 signaling in rat liver,” Journal of Molecular Medicine (Berlin, Germany), vol. 86, no. 11, pp. 1279–1285, 2008. View at: Publisher Site | Google Scholar
  49. H. H. Hsu, W. J. Ko, J. Y. Hsu et al., “Simvastatin ameliorates established pulmonary hypertension through a heme oxygenase-1 dependent pathway in rats,” Respiratory Research, vol. 10, no. 1, p. 32, 2009. View at: Publisher Site | Google Scholar
  50. W. H. Zhang, Y. J. Zhang, C. P. Liu, B. X. Yu, and W. X. Lu, “Simvastatin protects against the development of monocrotaline-induced pulmonary hypertension in rats via a heme oxygenase-1-dependent pathway,” Experimental Lung Research, vol. 37, no. 8, pp. 492–499, 2011. View at: Publisher Site | Google Scholar
  51. S. Dhivya, V. V. Padma, and E. Santhini, “Wound dressings - a review,” Biomedicine (Taipei), vol. 5, no. 4, p. 22, 2015. View at: Publisher Site | Google Scholar
  52. M. Matsumura, N. Fukuda, N. Kobayashi et al., “Effects of atorvastatin on angiogenesis in hindlimb ischemia and endothelial progenitor cell formation in rats,” Journal of Atherosclerosis and Thrombosis, vol. 16, no. 4, pp. 319–326, 2009. View at: Publisher Site | Google Scholar
  53. I. Baruscotti, F. Barchiesi, E. K. Jackson et al., “Estradiol stimulates capillary formation by human endothelial progenitor cells: role of estrogen receptor-{alpha}/{beta}, heme oxygenase 1, and tyrosine kinase,” Hypertension, vol. 56, no. 3, pp. 397–404, 2010. View at: Publisher Site | Google Scholar
  54. A. Parzonko, M. E. Czerwinska, A. K. Kiss, and M. Naruszewicz, “Oleuropein and oleacein may restore biological functions of endothelial progenitor cells impaired by angiotensin II via activation of Nrf2/heme oxygenase-1 pathway,” Phytomedicine, vol. 20, no. 12, pp. 1088–1094, 2013. View at: Publisher Site | Google Scholar
  55. Y. H. Chen, S. J. Lin, Y. L. Chen, P. L. Liu, and J. W. Chen, “Anti-inflammatory effects of different drugs/agents with antioxidant property on endothelial expression of adhesion molecules,” Cardiovascular & Hematological Disorders Drug Targets, vol. 6, no. 4, pp. 279–304, 2006. View at: Publisher Site | Google Scholar
  56. C. P. Lin, Y. H. Chen, H. B. Leu et al., “Anti-inflammatory strategies for homocysteine-related cardiovascular disease,” Frontiers in Bioscience (Landmark Edition), vol. 14, pp. 3836–3845, 2009. View at: Google Scholar
  57. A. Fortuno, G. S. Jose, M. U. Moreno, J. Diez, and G. Zalba, “Oxidative stress and vascular remodelling,” Experimental Physiology, vol. 90, no. 4, pp. 457–462, 2005. View at: Publisher Site | Google Scholar
  58. A. L. Furfaro, N. Traverso, C. Domenicotti et al., “The Nrf2/HO-1 axis in cancer cell growth and chemoresistance,” Oxidative Medicine and Cellular Longevity, vol. 2016, Article ID 1958174, p. 14, 2016. View at: Publisher Site | Google Scholar
  59. S. W. Ryter and A. M. Choi, “Targeting heme oxygenase-1 and carbon monoxide for therapeutic modulation of inflammation,” Translational Research: The Journal of Laboratory and Clinical Medicine, vol. 167, no. 1, pp. 7–34, 2016. View at: Publisher Site | Google Scholar

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