The Effect Of "Kidney-Brain Harmony" Electroacupuncture Therapy On The Surrounding Environment Of Dopaminergic Neurons In Parkinson's Disease Model Mice

Objective:Parkinson's disease(PD)is a disease caused by chronic degenerative changes in the central nervous system and poses a serious threat to the health of middle-aged and elderly people and their quality of life.In this study,electro-acupuncture therapy with brain-kidney and kidney acupuncture,acupuncture at Baihui,Shenshu,Taixi and other acupuncture points were used to activate brain and kidney,and to reinforce the kidney and brain to reconstruct the marrow to investigate the effect of electroacupuncture on PD mice.To protect against inflammatory injury and dopaminergic neurons in PD mice,to further explore the mechanism of action of electroacupuncture in preventing and treating Parkinson's disease,and to provide new ideas and clues for the treatment of this disease by Chinese medicine,and then to promote the use of electroacupuncture in clinical practice.Provide theoretical basis.Methods:48 SPF male 9.5-month-old C57BL/6J mice were selected to take natural light,freely drink water and feed,and were fed normally for 3 days.24 mice were injected intraperitoneally with MPTP(20 mg/kg·d).A sub-acute model of Parkinson's disease mice was established by continuous injection of 7 days at the same time every day.The electroacupuncture group began to use the SDZ-III Huake brand electro-acupuncture instrument to perform electro-acupuncture treatment after the completion of the modeling.The Huayin brand 0.25 x 13 mm needle was used,and the depth of acupuncture was 3 mm.The mice were alternately left and right,once/day,for a total of 28 days.The mice were tested for coordinated motor ability by suspension test,and electroacupuncture therapy was applied to Parkinson's disease model mice by immunohistochemical staining.The number of positive cells of tyrosine hydroxylase(TH)and glial fibrillary acidic protein(GFAP)and ionized calcium linker molecule(Iba-1)in substantia nigra and detection of electroacupuncture in Parkinson's disease mice using Elisa method The levels of interleukin-17(IL-17),interleukin-6(IL-6),tumor necrosis factor-a(TN Fa),and glial derived neurotrophic factor(GDNF)in the surrounding environment of dopaminergic neurons,Western blot was used to detect the expression of TH,GFAP and Iba-1 in substantia nigra.Results: 1.1 Observation of general behavior: Through observation,it was found that mice in the normal group were sensitive and had no limb disharmony.After 30 minutes of injection of MPTP,mice showed vertical tails and piloerections,increased jumps and twitches,and limb tremors occurred 1 hour later.After injection of MPTP for 7 days,mice showed movement disorders and evasion response was slow.After the treatment of electroacupuncture and Western medicine,the mice had flexible movements,limb coordination,limb tremors,stiffness,vertical tails,vertical hairs,jumps,twitches and other movements significantly reduced,and symptoms improved significantly.1.2 Hanging experiment: There was no significant difference in the hanging test scores of the mice before the modeling(P>0.05).After modeling,compared with the blank group,the hanging experiment of mice in each group changed significantly(P<0.05).After treatment,compared with the blank group,the suspension test in the model group was still significantly reduced(P <0.05);compared with the model group,the suspension experiment of the EA group and the western medicine group changed significantly(P <0.05).It was demonstrated that the mouse model could be maintained until the end of treatment,and the coordination ability of the treated mice improved.2.Immunohistochemical staining technique: compared with the blank group,the number of positive cells in the TH group was significantly reduced in the model group,and the number of GFAP and Iba-1 positive cells increased significantly(P<0.05),indicating that the model group of dopaminergic nerves The meta-damage was obvious and proved that the mouse model was successful.Compared with the model group,the number of positive cells in the TH group and the western medicine group significantly increased,and the number of GFAP and Iba-1 positive cells decreased significantly(P<0.05).3.The protein levels of TH,GFAP and Iba-1 in the substantia nigra of mice were detected by Western Blot method: compared with the blank group,the expression of TH protein in the substantia nigra of the model group decreased significantly(P<0.05),GFAP and Iba-1 The protein content was significantly increased(P<0.05);compared with the model group,TH protein expression was significantly increased in the substantia nigra of the EA group and the western medicine group(P<0.05),and the protein content of GFAP and Iba-1 was decreased.Significantly(P < 0.05).4.The levels of IL-17,IL-6,T N F-a,and GDNF in striatum,substantia nigra and serum of each group were detected by ELISA.Compared with the blank group,the levels of IL-17,IL-6 and TN Fa in the striatum,substantia nigra and serum of the model group increased significantly(P<0.05),and the content of GDNF decreased significantly(P<0.05).Compared with the model group,the content of IL-17,IL-6,TN Fa in the striatum,substantia nigra and serum of the electroacupuncture group and the western medicine group decreased(P<0.05),and the content of GDNF increased(P<0.05).Conclusion: Electroacupuncture therapy of “Kidney and Brain Communication”can inhibit excessive activation of astrocytes and microglia in the brain and reduce the proinflammatory effect of Inflammatory factors,and increase the neurotrophic effect of Nutrition factors,thus affecting the dopaminergic neuron environment.Play a role in the protection of dopaminergic neurons.