Research Of Echinacoside On Inflammation Mechanism Of MPTP Subacute Parkinson’s Disease Mouse Model

AIM:In application of MPTP subacute mouse model of Parkinson’s disease and observing protection of Echinacoside on DA neurons injury. Studying and discussing the mechanism of Echinacoside on inflammation caused by the MPTP subacute mouse model.Clarifying the possible molecular signaling pathways of Echinacoside on DA neurons protection.In aim to provide some beneficial ideas and methods of theory and practice for Chinese medicine treatments for PD.Methods:Choosing 10 week-old healthy wild type C57BL/6J male mice. Giving Echinacoside (ECH 30 mg·kg-1, once a day, from day 1 to the final day, continuously 16 days) or the same amount of saline, after 1 h granted MPTP (MPTP 30 mg-kg-1, once a day, from day 8 to day 12, continuously 5 days) or the same amount of saline subcutaneously to establish MPTP subacute Parkinson’s disease mouse inflammation model.Using locomotor activity, pole test and gait analysis to assess the dopaminergic motor neurons injury. Immunohistochemistry was taken for tyrosine hydroxylase (TH) and glial fibrillary acidic protein (GFAP) expression. The total numbers of TH-positive neurons and GFAP-positive cells in the substantia nigra pars compacts (SNpc) and ventral tegmental area (VTA) were obtained stereologically using the optical fractionator method. Enzyme-linked immunosorbent assay (ELISA) method was used to detect the inflammatory cytokines in the serum,including TNF-a(Tumor necrosis factor alpha),IFN-y(Interferon gama),IL-1β(Interleukin 1 beta).Protein expressions of ionized calcium binding adaptor molecule(IBA) 1, TNF-a, caspase-3, brain derived neurotrophic factor(BDNF), pro-BDNF, gial derived neurotrophic factor(GDNF), phosphorylated extracellular signal-regulated kinase 1/2(p-Erkl/2), Erkl/2, phosphorylated protein kinase B(p-AKT), AKT and p-mTOR (phosphorylated mammalian target of rapamycin) in the anatomical region of substantia nigra (SN) were tested by Western blotting.Results:Echinacoside increased total distance of MPTP injured mice in spontaneous activity (P<0.05) and shortened the total time in pole test (P<0.05).Howerver, the gait analysis shows no statistical difference (P> 0.05).Echinacoside significantly decreased 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced loss of TH-positive(TH+) neurons in the region of VTA (P<0.05) and SNpc (P<0.05). Attenuated GFAP reactivity was observed in answer to Echinacoside treatment in the region of VTA (P<0.05) and SNpc (P<0.05). The up-regulation of IBA 1 (P<0.05), pro-inflammatory TNF-a (P<0.05) and apoptotic protein caspase-3 (P<0.05) in the region of SN were inhibited by Echinacoside treatment. Echinacoside also decreased production of TNF-a (P<0.05), IFN-y (P<0.05), IL-1β(P> 0.05) in the serum. In addition, Echinacoside down-regulated pro-BDNF (P< 0.05), up-regulated expression of neurotrophic factor BDNF (P> 0.05),GDNF(P<0.05) and antiapoptotic factor p-ERK1/2/ERK1/2 (P< 0.05) in the region of SN. Echinacoside showed the trendency of up-regulating p-AKT/AKT (p> 0.05) and down-regulating p-mTOR protein expression (p>0.05).Conclusions:1. Echinacoside improved behavior performance of MPTP subacute Parkinson’s disease mouse model.2. Echinacoside plays a role in the protection of DA neurons of MPTP subacute Parkinson’s disease mouse model through anti-inflammation and anti apoptosis.3. Echinacoside inhibits nerve inflammation and enhances the DA neurons nutrition support by regulating the function of microglia and astrocytes.