Therapeutic Effect And Mechanism Of Zishen Yisui Recipe On Parkinson's Disease

[Background]Parkinson’s disease(PD)is a neurodegenerative disease commonly seen in the elderly.The course of the disease is progressively aggravated,incurable,and at risk of disability in the later stages,which seriously affects the quality of life of patients.Research has confirmed that about 80%of patients with PD have a decrease in striatal dopamine before the onset of symptoms,and 60%-70%of patients have degeneration of substantia nigra neurons.Magnetic Resonance Spectroscopy(MRS)technology can non-invasively observe the biochemical changes and tissue metabolism at the molecular level of human body,it provides a new technical method for PD patients in etiological research,disease diagnosis and differential diagnosis,disease progression monitoring,and disease prognosis evaluation.The metabolites in the brain tissue that 1H-MRS can detect are N-acetylaspartate(NAA),Choline(Cho),Creatine(Cr).The classic pathological changes of PD are the degeneration of dopaminergic neurons in the substantia nigra pars compacta,and the presence of Lewy bodies with a-synuclein(a-syn)as the key component in the remaining neurons.α-syn is a protein consisting of 140 amino acids and is located at the presynaptic end of neurons.The mutation or excessive accumulation of α-syn leads to α-syn oligomer formation,which can affect the normal physiological function of cells and even lead to cell death.Therefore,to explore the formation and degradation pathway of α-syn oligomers is becoming a new hotspot in basic research that can become a potential therapeutic target for synuclein disease.Chaperone mediated autophagy(CMA),which occurs in lysosomes,is one of the proteolytic pathways in mammalian cells that removes misfolded proteins.The a-syn oligomers can be degraded by the above pathway.Cathepsin D(Cath D)in lysosomes has been shown to prevent a-syn aggregation and toxicity.Molecular chaperones are important cytokines that mediate the formation and maintenance of the natural conformation of the helper proteins,ensuring proper folding and assembly of proteins.Heat shock protein 70(HSP70)belongs to the molecular chaperone,and it is currently found that it can inhibit the over-expression and aggregation of a-syn.PD treatment mainly includes drugs and surgical treatment.Except for some patients who meet the surgical Criteria and economic conditions,at present,PD is usually treated with oral western medicine.Both clinical and literature have shown that patients with PD may experience side effects such as diminished efficacy,switching phenomena,and dyskinesia when taking western medicine,and Western medicine is almost ineffective against non-motor symptoms of PD.Chinese medicine can assist Western medicine treatment,effectively reduce the side effects of Western medicine,delay the progression of the disease,and regulate the overall physical condition of patients,through theories such as syndrome differentiation and treatment.Zishenyisuifang(Chushizi,Shayuanzi,Gouqizi,Huangjing,Shudihuang,Roucongrong,Guiban)can both nourish the kidney yin and replenish the kidney yang,combine with the liver and spleen,and has the function of tonifying the kidney,filling the essence and benefiting the brain.It can also improve the patient’s movement and non-motor symptoms,such as sleep disorders,constipation,abnormal sweating,neuropsychiatric symptoms and other non-motor symptoms,improve the quality of life of patients.The Parkinson’s Disease TCM Quality of Life Scale(PD-QOL)was established under the guidance of the theory of traditional Chinese medicine with reference to an internationally recognized method for the development of scales.PD-QOL is a scale for the quality of life of PD patients and it is of great significance for further exploring the advantages of Chinese medicine in the treatment of PD.[Objective]To further investigate the effects of Zishenyisuifang on the quality of life of patients with PD,evaluate the effect of Zishenyisuifang on the activity of neurons in the substantia nigra of PD patients,provide imaging evidence for the treatment of PD,and to investigate the effect of Zishenyisuifang on the degradation of a-syn oligomers through the molecular chaperone-mediated autophagy(CMA)pathway in PD model mice and the possible mechanism of PD treatment,this research is to be divided into two parts,clinical and experimental.The clinical research used(PD-QOL)as an evaluation method to explore the effect of Zishenyisuifang on the quality of life of PD patients.Using magnetic resonance spectroscopy as a technical means,to investigate the effect of Zishenyisuifang on the expression of major metabolites NAA,Cho,and Cr in the substantia nigra of Parkinson’s disease patients.The purpose of this research is to verify the protective effect of Zishenyisuifang on substantia nigra dopaminergic neurons in PD patients.The experimental study used Cath D and Hsp70 as targets to explore the effect of Zishenyisuifang on the regulation of the lysosomal pathway in the brain of PD mice,and to clarify the mechanism of degradation of α-syn oligomers in the brain of PD mice.[Method]1 Clinical research methodsClinical studies included 50 patients with PD who met the Criteria.Based on anti-PD western medicine treatment,they were treated with Chinese medicine Zishenyisuifang for 3 months.At the time of enrollment and treatment for 3 months,PD general data collection,patient PD-QOL scale total scores,and Ⅱ,Ⅲ dimensions(PD-QOL-Ⅰ,PD-QOL-Ⅱ)scores were unified,UPDRS scores and Ⅱ,Ⅲ dimensions(UPDRS-Ⅱ,Ⅲ)scores,and Hoehn-Yahr classification,MDRSPD,nMS Quest,MoCA volume Tables,fatigue scales,sleepiness scales,sleep scales,constipation scales,etc.The values of NAA,Cr,and Cho in the substantia nigra region of PD patients were calculated by MRS,and the NAA/Cr and Cho/Cr ratios were calculated.By comparing the change of MRS data before and after treatment,the imaging basis of Zishen Miaofang in the treatment of PD was explored,and the correlation between the scores of the aforementioned scale and MRS data was analyzed.2 Experimental research methodsC57BL/6 male mice were randomly divided into normal group,model group,western medicine group and Chinese medicine group.PD mice models were established by continuous perfusion of low dose rotenone for a long time.The latter two groups were treated with metoprolol and Zishen Yishuifang after intervention.Autonomous activity test was used to observe the behavior of mice;immunohistochemistry was used to detect the expression of Cath D and HSP70 in the substantia nigra of mice.[Results]1 Clinical research resultsAfter treatment with Zishenyisuifang,PD-QOL scores and PD-QOL-Ⅰ,PD-QOL-Ⅱscores in PD patients were significantly lower than before treatment(P<0.01);UPDRS scores and UPDRS-Ⅱ,UPDRS-Ⅲ was significantly lower than before treatment(P<0.05 or P<0.01).At the time of enrollment,UPDRS-Ⅲ,UPDRS total score and PD-QOL-Ⅰ,PD-QOL-Ⅱ,PD-QOL total score of PD patients,and UPDRS-Ⅱ showed significant positive correlation with PD-QOL-Ⅰ and PD-QOL scores.(P<0.05);There was a significant positive correlation between MDRSPD score and PD-QOL total score(P<0.05).There was no significant correlation between nMS Quest,MoCA,Sleep Scale,Sleepiness Scale,Fatigue Scale,constipation Scale and PD-QOL Score and dimensions.The NAA/Cr value of the substantia nigra was significantly lower than that of the mild side of the disease(P<0.05).After treatment with Zishen Yishenfang,the above MRS data of the substantia nigra of PD patients were not significantly lower than before treatment(P>0.05).There was no difference in metabolites of the substantia nigra in patients with different genders,types,and Hoehn-Yahr grade PD patients.There was a significant positive correlation between age and severity of NAA/Cr in patients with PD(P<0.05),and there was no correlation between other substantia nigra metabolites MRS data and age of disease.There was a significant negative correlation between mild side NAA/Cr and UPDRS-Ⅱ+Ⅲ,PD-QOL-Ⅰ and PD-QOL-Ⅰ+Ⅱ in PD patients(P<0.05).There was a significant positive correlation between light-side Cho/Cr and UPDRS-Ⅲ,UPDRS-Ⅱ+Ⅲ,PD-QOL-Ⅰ and PD-QOL-Ⅰ+Ⅱ(P<0.05).The NAA content,NAA/Cr content in the severe side of PD patients were positively correlated with fatigue scores(P<0.05).There was a significant positive correlation between the Cho content and the MoCA score(P<0.05).There was no significant correlation between MRS data of bilateral substantia nigra metabolites and MDRSPD and nMS Quest in patients with PD.2 Experimental research resultsAfter Zishenyisuifang treatment,the number of times of spontaneous activity of mice in the Chinese medicine group and the western medicine group was significantly higher than that in the model group(P<0.05).There was no significant difference between the Chinese medicine group and the western medicine group.The expression of Cath D in the substantia nigra region of the model group was significantly lower than that of the blank group(P<0.05).The expression of Cath D in the substantia nigra of the western medicine group was significantly higher than that of the model group(P<0.01).The expression of Cath D in the substantia nigra of the Chinese medicine group was significantly higher than that of the model group(P<0.05).The expression of Cath D in the substantia nigra of the Chinese medicine group was significantly lower than that of the western medicine group(P<0.01).The positive expression of HSP70 in the substantia nigra of the model group was significantly lower than that of the blank group(P<0.05).The positive expression of HSP70 in the substantia nigra of the western medicine mice was significantly higher than that of the model group(P<0.05).The positive expression of HSP70 in the substantia nigra of the Chinese medicine mice was significantly higher than that of the model group(P<0.01).The positive expression of HSP70 in the substantia nigra of the Chinese medicine group was significantly higher than that of the western medicine group(P<0.01).[Conclusion]1.The PD-QOL(first draft)scale was used as an evaluation method to confirm that Zishenyisuifang can improve the patient’s quality of life by improving the patient’s motor symptoms and non-motor symptoms.The results of the research show that the improvement of the quality of life of Chinese medicine is closely related to the symptoms of exercise and non-exercise,and thus provides clinical support for the promotion of Parkinson’s disease treated by Chinese medicine.2.Using MRS as a technical means,can confirme that Zishenyisuifang can delay neuronal damage in substantia nigra and maintain neuronal activity by delaying the decline of NAA/Cr.So as to improve the patient’s motor symptoms and non-motor symptoms,effectively improve the quality of life of the patient.The results of the study provide evidence for the treatment of PD by Chinese medicine,and also provide support for MRS assessment of PD patients.3.Zishenyijingfang can enhance the CMA pathway by enhancing the expression of molecular chaperones Hsp70 and Cath D in brains of PD mice.This leads to the degradation of α-syn oligomers in the brain of PD mice.The results of this research can provide a modern scientific theoretical basis for the therapeutic mechanism of Zishenyisuifang on PD and the degradation of a-syn oligomers.