Abstract
Parkinson’s disease (PD) is a neurodegenerative disease with the pathological hallmark of reduced nigrostriatal dopamine. In traditional Chinese medicine (TCM) clinical practice, the nanopowder of Cistanche tubulosa has therapeutic effects on PD. To identify the therapeutic mechanism, this study tested the protective effect of different doses of MPP+-induced toxicity in MES23.5 cells using the MTT assay and in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mice (vehicles). Immunohistochemistry was used to assess cytomorphology and tyrosine hydroxylase (TH) expression. Behavioral tests in vehicles, high performance liquid chromatography (HPLC) tests in dopamine, immunohistochemistry and western blot analysis were used to detect the expression of TH, glial cell line-derived neurotrophic factor (GDNF) and its receptors. Our results demonstrated that the C. tubulosa nanopowder improved the viability of MPP+-treated cells, increased TH expression and reduced the number of apoptotic cells. It also increased Bcl2 protein expression and suppressed Bax protein expression in MPP+-treated cells in a dose-dependent manner. In addition, C. tubulosa nanopowder improved the behavioral deficits in vehicle mice, reduced the stationary duration of swimming, enhanced the ability for spontaneous activity and increased the expression of GDNF, the GDNF family receptor alpha (GFRα1) and Ret in cells of the substantia nigra (SN). Furthermore, the protein expression of GDNF, GFRα1 and Ret increased after treatment with different doses of C. tubulosa nanopowder, with a significant difference between the high-dose and vehicle groups. The protein expression of Bcl2 and Bax were similar in the in vivo and in vitro, which suggested that C. tubulosa nanopowder has anti-apoptotic effects in neurons.
Highlights
Parkinson’s disease (PD) is a common neurodegenerative disease occuring in elderly people with the pathological manifestations of loss of dopaminergic neurons in the substantia nigra (SN) due to degeneration
MPP+, MTT and glutamine were purchased from SigmaAldrich (Carlsbad, CA, USA); DMEM/F12 medium and fetal bovine serum were purchased from Gibco Co. (Life Technologies, Carlsbad, CA, USA); and MPTP, DA standard and homovanillic acid (HVA) standard were purchased from Sigma-Aldrich (Carlsbad, CA, USA). β-actin, Bax, Bcl2, glial cell line-derived neurotrophic factor (GDNF), GDNF family receptor alpha (GFRα1) and Ret antibodies were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA); 3,3’-diaminobenzidine (DAB) staining reagent kit was purchased from Fuzhou Maixin Biotech., Ltd. (Fujian, China); and SDS-PAGE gel sample preparation kit, ultrasensitive enhanced chemiluminescence (ECL) detection kit and bicinchoninic acid (BCA) assay were purchased from Beyotime Institute of Biotechnology (Beijing, China)
In the range of 200–400 nm scanning, ECH in C. tubulosa and VER in 330 nm had the maximum absorption peak, which appeared within 20 min
Summary
Parkinson’s disease (PD) is a common neurodegenerative disease occuring in elderly people with the pathological manifestations of loss of dopaminergic neurons in the substantia nigra (SN) due to degeneration. The severity of the disease has been shown to be correlated with dopamine (DA) neuronal cell loss in the SN, which is consistent with the view that the neurodegenerative. Among all of the neurotrophic factors that protect and promote the repair of dopaminergic neurons, GDNF has the strongest effects (Hong et al, 2008; Rangasamy et al, 2010; Allen et al, 2013). The increased incidence of neuronal apoptosis and reduced protective effects of neurotrophic factors, potentially triggered by various pathological factors, underlie the degeneration of dopaminergic neurons (Holden et al, 2006)
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