Objective: To elucidate the effect of P2X4 signal axis on iron metabolism in substantia nigra（SN）of male rats with Parkinson’s disease（PD）successfully induced by 6-hydroxydopamine (6-OHDA).

Background: Related studies have confirmed that the ATP-P2X4 signal axis can promote the rearrangement of its subunits and the opening of iron channels. The interaction and deposition of iron ions with α-synuclein are the core links of dopaminergic neuron degeneration. Because abnormal iron metabolism is one of the important mechanisms for the occurrence and development of PD, we conclude that the P2X4 gene axis is likely to affect iron metabolism in PD animal models, and eventually lead to the occurrence and development of PD.

Method: Male rats were randomly divided into control group, PD group, P2X4-gene virus group, P2X4-gene unloaded virus group, P2X4-PI+6-OHDA group and P2X4-NC+6-OHDA group. Brain stereotactic instrument was used to inject the corresponding grouped drugs into the left SN of rats. A behavioral test was performed 2 weeks after the modeling was completed to select the qualified rat models, and the initiation and balance ability of each group of rats were further evaluated by a balance bar experiment. The brain of the qualified rat models were decapitated, and the brain tissue was taken away and preserved after related treatment. Immunofluorescence staining and Western blot method were used to detect the number of (TH) positive dopaminergic neurons, and the expression levels of protein in P2X4 purinergic receptor (P2X4R), Divalent metal-ion transporter-1(DMT1) and Ferroportin1 (FPN1).

Results: 1. The result of immunofluorescence staining showed that the number of TH positive dopaminergic neurons in the left SN of PD group and P2X4-NC+6-OHDA group were significantly lower than that of the control group（P＜0.01; P＜0.001）. The number of TH positive dopaminergic neurons of P2X4-PI+6-OHDA group were significantly lower than that of P2X4-NC+6-OHDA group（P＜0.01）. 2. The result of Western blot suggested that compared with the control group, the expression of P2X4R, DMT1 in the left SN of PD group and P2X4-NC+6-OHDA group were increased, but FPN1 decreased(P＜0.05). Compared with P2X4-NC+6-OHDA group, the expression of P2X4R, DMT1 in left SN of P2X4-PI+6-OHDA group increased and FPN1 decreased（P＜0.05）.

Conclusion: The overexpression of P2X4 gene can up-regulate the expression of DMT1 and down-regulate the expression of FPN1 in the SN.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/effect-of-p2x4-signal-axis-on-iron-metabolism-in-parkinsons-disease-animal-model/