Biochemical and Biophysical Research Communications
Effects of pergolide mesylate on transduction efficiency of PEP-1-catalase protein
Research highlights
► We studied effects of pergolide mesylate (PM) on in vitro and in vivo transduction of PEP-1-catalase. ► PEP-1-catatase inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation. ► PM enhanced the transduction of PEP-1-catalase into HaCaT cells and skin tissue. ► PM increased anti-inflammatory activity of PEP-1-catalase. ► PM stimulated therapeutic action of anti-oxidant enzyme catalase in oxidative-related diseases.
Introduction
Reactive oxygen species (ROS) such as hydroxyl radicals, superoxide anion and hydrogen peroxide, are produced from various normal cellular processes and lead to cell damage, cell death and consequently, several diseases including inflammation, cancer, allergy, and ischemia [1], [2], [3]. Anti-oxidant enzymes, like superoxide dismutase (SOD), catalase, and glutathione peroxidase play important roles in decreasing overproduced ROS, maintaining a balance of the redox state and protecting cells against numerous sources of oxidative damage [4], [5], [6], [7]. This suggests that these anti-oxidant enzymes may be useful as therapeutic molecules. However, delivering several proteins with therapeutic potential into cells is difficult due to their size and biochemical properties, making it problematic to utilize such proteins as therapeutic drugs [8].
Many researchers have reported that therapeutic proteins fused with protein transduction domains (PTD) such as Tat and PEP-1 can be efficiently delivered into cells [9], [10], [11], [12], [13], [14], [15], [16]. Also, we constructed the expression vectors of PEP-1-catalase, PEP-1-rpS3, and Tat-SOD fusion proteins. PEP-1-catalase fusion protein was reported to be rapidly transduced into astrocytes and gerbil brain and protected neuronal cells against ischemic damage [17]. The transduction of Tat-SOD into pancreatic beta cells improved the diabetic status of mice in a diabetic mouse model [18]. In addition, transduced PEP-1-rpS3 prevented TPA-induced skin inflammation in a mice model by inhibiting the expression of pro-inflammatory mediator and cytokines such as cyclooxygenase-2 (COX-2), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) [19].
Dopamine receptor agonists are divided into ergolines and non-ergolines. Ergolines are derived from ergot alkaloids and include bromocriptine, pergolide, lisuride and cabergoline. They bind to dopaminergic D1 receptors, adrenergic and 5-hydroxytryptamine receptors as well as dopaminergic D2 receptors. Pergolide mesylate (PM) is a synthetic ergoline derivative and is used in the treatment of Parkinson’s disease (PD) [20]. Also, some studies have reported that pergolide has free radical scavenging activity and an inhibitory activity on phospholipid peroxidation in rat brain and induces Cu,Zn-SOD activity in rat striatum [21], [22]. It exerts its neuroprotective activity by interfering with NF-κB translocation in neuronal cell line exposed to H2O2[23], [24], [25]. However, the effect of PM on the cellular membrane conformation and its anti-inflammatory effect on skin cells and tissue are still unknown.
Therefore, in this study, we investigated whether PM could enhance the transduction efficiency and thereby the anti-inflammatory effects of PEP-1-catalase. Subsequently we found that PM enhanced the transduction efficiency of PEP-1-catalase in vitro and in vivo, and that in turn increased the efficiency with which it inhibited the expression levels of anti-inflammatory mediators and cytokines.
Section snippets
Materials
PM was purchased from Sigma–Aldrich (St. Louis, MO, USA). Primary antibodies against COX-2, iNOS, IL-6, IL-1β and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All other chemicals and reagents were obtained from Sigma–Aldrich (MO, USA) unless otherwise stated.
Cell culture and MTT assay
HaCaT human keratinocytes were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and antibiotics (100 μg/ml streptomycin and 100 U/ml penicillin) and kept at 37
Effect of PM on in vitro and in vivo transduction efficiency of PEP-1-catalase
Protein transduction domains (PTDs) such as Tat and PEP-1 are able to deliver exogenous macromolecules to cells and animals [9], [15], [16], [28]. Several studies have suggested that PTD fusion therapeutic molecules may be useful drugs against various disorders [29], [30], [31], [32]. Anti-oxidant enzymes, such as superoxide dismutase (SOD), catalase and glutathione peroxidase, are known to maintain the cellular redox balance and work as a cellular defenses mechanism. Earlier we reported that
Acknowledgments
This work was supported by the Priority Research Centers Program grant (2009-0093812) and by the Regional Research Universities Program/Medical & Bio-material Research Center grant through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology.
References (40)
- et al.
Oxidative stress in the pathogenesis of skin disease
J. Invest. Dermatol.
(2006) Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology
Toxicology
(2000)- et al.
Liposome-mediated augmentation of superoxide dismutase in endothelial cells prevents oxygen injury
J. Biol. Chem.
(1983) Targeting of superoxide dismutase and catalase to vascular endothelium
J. Control Release
(2001)- et al.
Protection of cultured endothelial cells from hydrogen peroxide-induced injury by antibody-conjugated catalase
Biochim. Biophys. Acta
(1987) - et al.
Bioavailability and transport of peptides and peptide drugs into the brain
Peptides
(1997) - et al.
Protein transduction: unrestricted delivery into all cells?
Trends Cell Biol.
(2000) - et al.
Protein transduction technology
Curr. Opin. Biotechnol.
(2002) - et al.
Transduced human PEP-1-catalase fusion protein attenuates ischemic neuronal damage
Free Radic. Biol. Med.
(2009) - et al.
HIV-1 Tat-mediated protein transduction of Cu,Zn-superoxide dismutase into pancreatic beta cells in vitro and in vivo
Free Radic. Biol. Med.
(2004)
Transduced PEP-1-ribosomal protein S3 (rpS3) ameliorates 12-O-tetradecanoylphorbol-13-acetate-induced inflammation in mice
Toxicology
Pergolide scavenges both hydroxyl and nitric oxide free radicals in vitro and inhibits lipid peroxidation in different regions of the rat brain
Brain Res.
Pergolide protects SH-SY5Y cells against neurodegeneration induced by H2O2
Eur. J. Pharmacol.
A novel mechanism for pergolide-induced neuroprotection: inhibition of NF-kB nuclear translocation
Biochem. Pharmacol.
Topical transduction of superoxide dismutase mediated by HIV-1 Tat protein transduction domain ameliorates 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice
Biochem. Pharmacol.
Preparation and fuctional analysis of recombinant protein transduction domain-metallothionein fusion proteins
Biochimie
Protein transduction domain-mediated delivery of QBP1 suppresses polyglutamine-induced neurodegeneration in vivo
Mol. Ther.
Human PEP-1-ribosomal protein S3 protects against UV-induced skin cell death
FEBS Lett.
Hydroxyl radical and superoxide dismutase in blood of patients with Parkinson’s disease: relationship to clinical data
J. Neurol. Sci.
Role of NF-kB in the apoptotic-resistant phenotype of keratinocytes
J. Biol. Chem.
Cited by (2)
Enhancement of anti-inflammatory activity of PEP-1-FK506 binding protein by silk fibroin peptide
2012, Journal of Microbiology and Biotechnology
- 1
These authors equally contributed to this work.