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This study aimed to explore the protective effects of  Wenshen-Yanggan  decoction on dopaminergic (DA) neuron injury in a rotenone-induced mouse model with chronic Parkinson's disease (PD) and explore its mechanism of action. Ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to measure the content of six main components in the  Wenshen-Yanggan  decoction. The chronic PD mouse model was established by treating 10-month-old healthy wild C57BL/6 male mice with rotenone 30 mg/kg/day for 28 days in succession. The pole test and rotarod test were applied to detect the rescue effect of  Wenshen-Yanggan  decoction in high, medium, and low dosages, respectively, on PD-like behaviors in mice with chronic PD. The protective effect of  Wenshen-Yanggan  decoction on the mesencephalic nigrostriatal DA neuron injury was determined employing tyrosine hydroxylase (TH) immunofluorescence staining. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the inflammatory cytokines in serum, including TNF- α  (tumor necrosis factor-alpha), IFN- γ  (interferon gamma), NF- κ B (nuclear factor kappa-B), and IL-1 β  (interleukin-1 beta). Western blotting was performed to quantify the expression of phosphorylated  c -Jun N-terminal kinase ( p -JNK), cleaved caspase-3, B-cell lymphoma-2 (Bcl-2), and NF- κ B in the brain. Our results showed that the  Wenshen-Yanggan  decoction in high, medium, and low dosages reduced the turning time of mice ( P  < 0.01,  P  < 0.01,  and  P  < 0.05). The high and medium dosages shortened the total climbing time of PD mice in the pole test ( P  < 0.01 and  P  < 0.05). Meanwhile, the high, medium, and low dosages increased the rod-standing time of PD mice in the rotarod test ( P  < 0.01,  P  < 0.05,  and  P  < 0.05). Besides, the decoction reversed the decrease in TH-positive neurons induced by rotenone, upregulated TH protein expression, and downregulated the  α -syn expression in the PD model. Moreover, the decoction in high dosage significantly inhibited the expression of  p -JNK, cleaved caspase-3, and NF- κ B in the midbrain of PD mice ( P  < 0.05,  P  < 0.05,  and  P  < 0.01), upregulated the expression of Bcl-2 ( P  < 0.05), and decreased the content of TNF- α , IFN- γ , NF- κ B, and IL-1 β  in the serum ( P  < 0.001,  P  < 0.001,  P  < 0.001,  and  P  < 0.001). Taken together, the  Wenshen-Yanggan  decoction could protect mice from rotenone-induced chronic PD, which might be related to the reduction of the DA neuron apoptosis via suppressing the inflammatory reaction and the neuronal apoptosis pathway.

Effect of Wenshen-Yanggan Decoction on Movement Disorder and Substantia Nigra Dopaminergic Neurons in Mice with Chronic Parkinson’s Disease