Abstract

To investigate the effect of Da-Bu-Yin-Wan and Qian-Zheng-San (DBYW and QZS) on mitochondrial mass in Parkinson’s disease (PD) cell model induced by 1-Methyl-4-phenylpyridinium Ion (MPP+). The SH-SY5Y cell was selected and treated with MPP+. The PD model was intervened with DBYW and QZS. CCK-8 method was used to detect the survival rate of cells in each group. Mitochondria was labeled by mitoTracker®Red CMXRos probe and observed by laser scanning confocal microscope, and ImageJ software was used to process images and measure mitochondrial form factors; Tetramethylrhodamine methyl ester was used to detect mitochondrial membrane potential (ΔΨm); Luciferase method was used to detect cellular ATP levels; Western-Blot technique was applied to detect the expression levels of Parkin protein, and the expression levels of Mfn1, Mfn2, OPA1, Drp1, and Fis1. We found that DBYW and QZS treatment significantly increased the cell survival rate, form factor (F-factor), mitochondrial activity and ΔΨm after MPP+ treatment, while the increase of ATP levels was not significant. In addition, the results of western blot analysis showed that the MPP+ induced increase in the expression of Drp1 and Fis1, as well as decrease in Parkin, Mfn1, Mfn2, and OPA1 were all partially revised by DBYW and QZS. In summary, our data strongly suggested that DBYW and QZS treatment can exert protective effects against PD related neuronal injury through regulation the homeostasis between mitochondrial fission and fusion.

Highlights

  • Parkinson’s disease (PD) is a progressive degenerative disease of the central nervous system caused by degeneration of dopaminergic neurons in the substantia nigra (Braak et al, 2004)

  • The results showed that compared with the control group, the expression of Parkin (P < 0.01), Mfn1 (P < 0.05), Mfn2 (P < 0.01) and OPA1 (P < 0.01) in the model group decreased significantly, whereas the expression of the two fission proteins, both dynamin-related protein 1 (Drp1) and Fis1 were increased by MPP+ treatment (P < 0.01)

  • Compared with the normal control group, the ATP levels in the model group and the group with the Chinese herbal compound decreased significantly (P < 0.05), and the model group decreased 22.9% compared with the normal control group, In this experiment, the selected SH-SY5Y cell strain of human neuroblastoma is characterized by its cell morphology, physiology and biochemical characteristics similar to normal neurons, rapid reproduction, and many properties of dopaminergic neurons

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Summary

Introduction

Parkinson’s disease (PD) is a progressive degenerative disease of the central nervous system caused by degeneration of dopaminergic neurons in the substantia nigra (Braak et al, 2004). It is mainly characterized by resting tremors, muscle rigidity, slow movement and disorder of postural reflexes, and often accompanied by symptoms of mental disorders, leading to changes in cognitive function, mental state abnormalities (Kehagia, 2016). DBYW and QZS Ameliorate Mitochondrial Dynamics morphology, dysfunction and unbalanced dynamics were closely related to the pathogenesis of PD (Bose and Beal, 2016; Giannoccaro et al, 2017). A large number of studies have shown that mitochondrial dyskinesia is closely related to various neurodegenerative diseases, especially PD

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