Research ReportNeurotrophic and neurorescue effects of Echinacoside in the subacute MPTP mouse model of Parkinson's disease
Introduction
Parkinson's disease (PD) is characterized by progressive dopaminergic neuronal cell death in the substantia nigra, resulting in severe motor deficits. In contrast to neuroprotective strategies, which aim to slow degeneration and thus must be initiated before significant damage has occurred, neurorescue or restorative therapies aim to obliterate neuronal deficits and degeneration after the initiation of impairments. Neurotrophic factors (NTFs) are secreted proteins that regulate the survival, functional maintenance and phenotypic development of neuronal cells, in particular glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) are widely recognized as potent survival factors for the dopaminergic neurons and the nigrostriatal pathway that degenerate in PD. While the neuroprotective or neurorescue activities of these NTFs have been demonstrated in a number of in vitro and in vivo experiments (Peterson and Nutt, 2008), their application as therapeutic agents for PD has been hampered by the difficulties in delivering them into the brain because the macromolecule peptides cannot pass the brain blood barrier (BBB). Thus clinical trials testing treatment of PD using either surgery or gene therapy must solve the problem of efficient delivery and expression of NTFs (Dietz et al., 2006, Torp et al., 2006). As an alternative to such invasive approaches, a potential strategy is to stimulate the endogenous expression of NTFs. Recently, several non-peptide NTF inducers, such as XIB4035 and PYM50028, have been shown to attenuate the loss of dopaminergic neurons and might have beneficial effects in the treatment of PD (Tokugawa et al., 2003, Visanji et al., 2008). These low molecular weight agents can be administered systemically, and they would thus overcome most of the problems associated with the efficacy and safety of delivering NTFs into the brain by such methods as induction of NTF expression by viral vectors or with the use of encapsulated NTF producing cells (Bespalov and Saarma, 2007). Thus, molecules that either induce the endogenous expression of NTFs or enhance their signaling have been given increasing attention as alternative therapeutic options for PD.
Echinacoside (ECH, Fig. 1) is one of the principal constituents of phenylethanoid glycosides (PhGs) extracted from a famous traditional Chinese medicine, Herba Cistanches (the stems of Cistanche deserticola, Cistanche salsa or Cistanche tubulosa). It is also the main phenolic component in the roots of Echinacea angustifolia and E. pallida (Pellati et al., 2004), which are widely used in Europe and North America for their immunoregulation properties (Hostettmann, 2003).As a polyphenol natural product ECH has various bioactivities, such as powerful antioxidant (Sloley et al., 2001, Dalby-Brown et al., 2005), free radical scavenging and reduce the amount of NO radical generated by activated macrophages (Xiong et al., 2000) so that it can protect against hepatotoxicity (Wu et al., 2007) and acute lung injury in rats (Zhang et al., 2007, Xiong et al., 1998). It also has anti-inflammatory, antitumor, anti-aging and improving learning memory activities (reviewed by He et al., 2009). Several studies have reported ECH has anti-apoptotic and neuroprotective properties in neurodegenerative disease, for example, ECH shows significant neuroprotective effects on H2O2 injured PC-12 cells and rescues SH-SY5Y neuronal cells from TNF-α induce apoptosis through the mitochondrial pathway (Kuang et al., 2009, Deng et al., 2004). In vivo the compound has protected dopaminergic neurons in 6-hydroxydopamine (6-OHDA)-lesioned rats through attenuating the diminution of dopamine (DA) and its intermediates (Chen et al., 2007a, Chen et al., 2007b). Another study reported that ECH had neuroprotective effects and behavioral improvement in the mouse model of MPTP-induced dopaminergic neuronal damage, while further investigation revealed that ECH induced down-regulation of caspase-3 and caspase-8 activation in MPP+-induced apoptosis of cerebellar granule neurons (Geng et al., 2007). But these studies only investigated the anti-apoptotic or neuroprotective roles of ECH by the method of pre-treatment. The questions of whether ECH affects other dopaminergic neuronal prosurvival and death-related factors (such as NTFs and the Bcl-2 family) and whether it has neurorescue effects have not yet been addressed. Through reviewing the sparse literature on NTF induction by herbs or herbal compounds, we found that Rehmannia glutinosa and catalpol, two herbs with similar functions to Cistanches according to the traditional Chinese medical theory, could induce GDNF gene and protein expressions both in vitro and in vivo then attenuate MPTP-induced neuronal degeneration of nigral-striatal dopaminergic pathway (Yu et al., 2006, Xu et al., 2010). We therefore hypothesized that ECH may also have analogous efficacy. On the other hand the subacute MPTP mouse model is a quite popular regimen which involves one injection of 30 mg/kg daily for five consecutive days. This regimen causes apoptosis and depletes striatal dopamine by 40–50% in young adult C57BL/6 mice, and the dopaminergic lesion stabilizes by 21 days after MPTP administration (Tatton and Kish, 1997, Jackson-Lewis and Przedborski, 2007). So our current study chooses this model and investigates the effects of ECH on expressions of GDNF and BDNF, two of the most potent NTFs associated with dopaminergic neurons survival, and it further demonstrates the neurorescue and anti-apoptotic capabilities of this compound after subacute MPTP lesion induction in a mouse model of PD.
Section snippets
ECH improves gait dysfunctions in a subacute MPTP mouse model of PD
Shortened stride length is one of the chief characteristics of abnormal gait in PD. The results of our study demonstrate a significant decrease in forelimb and hindlimb stride lengths in the MPTP-lesioned mice treated with vehicle (saline) at the 9th day after the last administration of MPTP, i.e., the 7th day of vehicle treatment, compared with the normal control group (p = 0.023, 0.014 , Figs. 2A, B). Meanwhile, the stride lengths of mice in the high-dose ECH (30 mg/kg/day) treatment group is
Discussion
In the present study, we demonstrate for the first time that ECH, a monomer extracted from a traditional Chinese herb, has neurorescue activities in subacute MPTP mouse model of PD as well as the potential to induce the expression of NTFs at both transcription and protein levels. After 14 days of oral administration, ECH demonstrated multifunctional effects on rescuing dopaminergic neurons function, relieving the reduction of DAT, inhibiting the growth in number of apoptosis cells in SNpc and
Animal protocols
All procedures were approved by the Animal Ethical Committee of Zhongshan Hospital Fudan University and carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.
Experiments were conducted using male C57BL/6 mice purchased from the Shanghai Slac Laboratory Animal Company (Shanghai, China) at 10 weeks of age and weighing 24–26 g. The animals were maintained in standard conditions (12 ⁄ 12 h light ⁄ dark cycle, 21 ± 2 °C, and relative humidity of
Acknowledgments
This work was supported by the Scientific and Technical Supporting Program, the Ministry of Science and Technology of China (No. 2006BAI04A11-3) and the Medical Leader Sponsorship by Shanghai Municipal Government (No. 2007-057).We thank Professor Fang Huang and Professor Danian Zhu for their guidance on the experiments and manuscript.
References (55)
- et al.
Effects of GDNF pretreatment on function and survival of transplanted fetal ventral mesencephalic cells in the 6-OHDA rat model of Parkinson's disease
Brain Res.
(2009) - et al.
GDNF family receptor complexes are emerging drug targets
Trends Pharmacol. Sci.
(2007) - et al.
An assessment of the validity of densitometric measures of striatal tyrosine hydroxylase-positive fibers: relationship to apomorphine-induced rotations in 6-hydroxydopamine lesioned rats
J. Neurosci. Meth.
(1990) - et al.
Echinacoside prevents the striatal extracellular levels of monoamine neurotransmitters from diminution in 6-hydroxydopamine lesion rats
J. Ethnopharmacol.
(2007) - et al.
Echinacoside rescues the SHSY5Y neuronal cells from TNFalpha-induced apoptosis
Eur. J. Pharmacol.
(2004) - et al.
Application of a blood–brain-barrier-penetrating form of GDNF in a mouse model for Parkinson's disease
Brain Res.
(2006) - et al.
A simple method to measure stride length as an index of nigrostriatal dysfunction in mice
J. Neurosci. Methods
(2002) - et al.
Neuroprotective effects of echinacoside in the mouse MPTP model of Parkinson's disease
Eur. J. Pharmacol.
(2007) - et al.
Neurotoxicity of MPTP to migrating neuroblasts: studies in acute and subacute mouse models of Parkinson's disease
Neurotoxicology
(2008) - et al.
Transplantation of human amniotic cells exerts neuroprotection in MPTP-induced Parkinson disease mice
Brain Res.
(2008)
Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method
Methods
Involvement of neurotrophic factors in aging of noradrenergic innervations in hippocampus and frontal cortex
Neurosci. Res.
Early signs of neuronal apoptosis in the substantia nigra pars compacta of the progressive neurodegenerative mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model of Parkinson's disease
Neuroscience
Analysis of phenolic compounds and radical scavenging activity of Echinacea spp
J. Pharm. Biomed. Anal.
Treatment of Parkinson's disease with trophic factors
Neurotherapeutics
Mouse model of Parkinsonism: a comparison between subacute MPTP and chronic MPTP/probenecid treatment
Neuroscience
Driving GDNF expression: the green and the red traffic lights
Prog. Neurobiol.
Activation of tyrosine kinase receptor signaling pathway by rasagiline facilitates neurorescue and restoration of nigrostriatal dopamine neurons in post-MPTP-induced parkinsonism
Neurobiol. Dis.
The synthetic ceramide analog L-PDMP partially protects striatal dopamine levels but does not promote dopamine neuron survival in murine models of parkinsonism
Brain Res.
In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining
Neuroscience
Detection of behavioral impairments correlated to neurochemical deficits in mice treated with moderate doses of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine
Exp. Neurol.
XIB4035, a novel nonpeptidyl small molecule agonist for GFRalpha-1
Neurochem. Int.
Protective effects of echinacoside on carbon tetrachloride-induced hepatotoxicity in rats
Toxicology
Inhibition of nitric oxide by phenylethanoids in activated macrophages
Eur. J. Pharmacol.
Catalpol attenuates MPTP induced neuronal degeneration of nigral-striatal dopaminergic pathway in mice through elevating glial cell derived neurotrophic factor in striatum
Neuroscience
Rehmannia glutinosa induces glial cell line-derived neurotrophic factor gene expression in astroglial cells via cPKC and ERK1/2 pathways independently
Pharmacol. Res.
Chinese herbs and herbal extracts for neuroprotection of dopaminergic neurons and potential therapeutic treatment of Parkinson's disease
CNS Neurol. Disord. Drug Targets
Cited by (95)
Biological phenethyl glycosides from plants
2023, Privileged Scaffolds in Drug DiscoveryEchinacoside prevents hypoxic pulmonary hypertension by regulating the pulmonary artery function
2020, Journal of Pharmacological SciencesCitation Excerpt :It is characterized by a pulmonary artery pressure higher than 30 mmHg, with various factors contributing to the pathogenesis of the disease, leading to an abnormal constriction of pulmonary arteries and inducing proliferation of pulmonary arterial smooth muscle cells (PASMCs).5 Echinacoside (ECH) (Fig. 1) is a phenylethanoid glycoside from the Tibetan herb Lagotis brevituba Maxim and Cistanche tubulosa.6,7 Our previous findings indicated that ECH induces the vasorelaxation of rat pulmonary artery, and also inhibits hypoxia induced-proliferation of rat PASMCs in a concentration-dependent manner.8,9
Echinacoside selectively rescues complex I inhibition-induced mitochondrial respiratory impairment via enhancing complex II activity
2019, Neurochemistry InternationalSerum level of brain-derived neurotrophic factor in Parkinson's disease: a meta-analysis
2019, Progress in Neuro-Psychopharmacology and Biological PsychiatryCitation Excerpt :Reduced BDNF expression in the substantia nigra in animal results in similar symptoms of PD (Baquet et al., 2005; Porritt et al., 2005), whereas artificial increase of BDNF in experimental animal models of PD rescues degenerating dopamine neurons (Isacson et al., 1995; Levivier et al., 1995; Tsukahara et al., 1995; Hung and Lee, 1996; Murer et al., 2001). Consistently, a lot of improving treatments of PD are accompanied by BDNF enhancement (Okazawa et al., 1992; Bousquet et al., 2009; Naoi and Maruyama, 2009; Gyarfas et al., 2010; Li et al., 2010a; Li et al., 2010b; Zhao et al., 2010). All these results suggest that dysfunction of BDNF is involved in the pathogenesis of PD and provide cautious optimism that BDNF could act as a therapeutic agent of PD.