A Study On The Effect Of Zhimuning On PD Mitophagy And A Systematic Review Of Traditional Chinese Medicine In The Treatment Of PD With Depression

BackgroundParkinson's disease(PD)is the most prevailing neurodegenerative disease,with the progressive neuronal degeneration and loss as the main pathological features.PD mainly presents motor symptoms along with non-motor symptoms,including rest tremor,gait abnormalities,muscle rigidity,anxiety,depression,constipation and cognitive impairment.Currently,the etiology and pathogenesis of PD remain elusive.Multiple lines of evidences indicate that mitochondrial dysfunction is one of considerable pathogenic mechanisms of PD.Aberrant mitochondrial morphology is closely related to mitochondrial fission and dysregulation of mitochondrial autophagy(mitophagy).In recent years,it has been found that the inhibitor of dynamin-related protein 1(Drp 1)can efficaciously prevent mitochondrial fission and decrease the occurrence of mitochondrial autophagy.Traditional Chinese medicine(TCM)possess a significant advantage in the treatment of PD.Accumulating studies suggest that mangiferin can traverse the blood-brain barrier to diminish free radicals,reduce neuroinflammation,and restore mitochondrial bioenergy,thereby exerting neuroprotective effects.However,there is no report concerning the effects of mangiferin on mitochondrial morphology and mitophagic pathways in PD models.In this work,we investigated the mechanisms of mangiferin on Drpl-mediated mitophagy in vivo.Furthermore,literature research indicated that most PD patients suffered from depressive symptom contributing to a poorer prognosis and a lower quality life for patients.And TCM formulas combined with selective serotonin reuptake inhibitors(SSRIs)have an effective impact on PD patients with depression.We performed a systematic review to evaluate the efficacy of TCM formulas on depressed PD.ObjectiveExperimental research:1.We aim to investigate the relationship between the pathogenesis of Parkinson's disease and Drpl-mediated mitophagy.2.We aim to evaluate the pharmacological effects of mangiferin on the mice of PD model.Evidence-based medicine research:We aim to evaluate the efficacy of TCM formulas combine with SSRIs for depressed PD in a systematic review and meta-analysis.MethodsExperimental research:1.C57BL/6 adult mice were divided into three groups:the control group(saline-injected group),the model group(MPTP-injected group),and the dynasore-injected group.Firstly,three groups were injected intraperitoneally with an equal volume of 0.9%saline or MPTP for consecutive 5 days,followed by dynasore injection in the dynasore-treated group.Secondly,we performed open filed test,pole test,and rotarod test to detect the effects of dynasore on the motor and non-motor symptoms of mice.Then all mice were sacrificed for further analysis.Immunohistochemistry assessed the expression of tyrosine hydroxylase in tissue sections;immunofluorescence evaluated the expression and localization of Drpl protein in the striatum;transmission electron microscopy observed the mitochondrial morphology in the striatum;immunoblotting measured Drp1 protein content and mitochondrial autophagy-related proteins,including PINK1.Parkin,NIX,BNIP3,FUNDC1,p62 and LC3B.2.Mice were randomly divided into five groups:the control group(0.9%saline-injected group),the model group(MPTP-injected group),mangiferin low-dose group,mangiferin middle-dose group,and mangiferin high-dose group.Firstly,three groups were injected intraperitoneally with an equal volume of 0.9%saline or MPTP for consecutive 5 days,followed by the injection of mangiferin.Behavioral tests were performed after the last treatment of mangiferin.According to the results of behavioral tests,further molecular analysis was conducted including immunohistochemistry,immunofluorescence,transmission electron microscopy,and protein immunoblotting.Evidence-based medicine research:We searched literatures from databases of PubMed,Web of Science,Embase,The Cochrane Library,Chinese Biomedical Literature,CNKI,Vip,and Wanfang.And we included randomized controlled trials investigating the efficacy of TCM formulas combined with SSRIs on depressed PD patients.The search method was based on a combination of MeSH subject words and free words.The search terms included"depress","parkinson*","traditional Chinese medicine",and "Chinese medicine" in English;and contained "Chinese medicine","Parkinson's disease","depression",and "randomized controlled trials" in Chinese.The main observations were the efficacy,Hamilton Depression Scale(HAMD)score,UPDRS II and UPDRS III scores.Literature quality was evaluated using Cochrane Manual criteria,involving randomized grouping scheme,group concealment,blinding,data integrity,selective reporting,and other bias.The fixed-effect model was applied when the heterogeneity of included studies was less than 50%.The Egger's test and Begg's test were conducted to measure the publication bias,and the funnel plots were performed.ResultsExperimental research:1.The results showed that Drpl inhibitor could ameliorate partial motor dysfunction of PD-related model mice(p<0.05);immunohistochemical results showed that Drpl inhibitor could up-regulate tyrosine hydroxylase expression(p<0.01);transmission electron microscopy showed that inhibition of Drpl expression could protect mitochondrial microstructure in the striatum;the results of immunoblotting showed that inhibition of Drp1 expression could up-regulate the expression of PINK1 and Parkin proteins in mitochondria(p<0.05).but there was no significant difference on the expression of NIX and BNIP3 proteins.2.The results showed that mangiferin could improve the motor dysfunction of PD-related model mice(p<0.05);immunohistochemical results showed that the expression of tyrosine hydroxylase could be significantly up-regulated by the medium and mangiferin high-dose groups(p<0.01);transmission electron microscopy observed that mangiferin middle-and high-dose groups of Drp1 could protect the mitochondrial ultrastructure in the striatum of mice;the results of immunofluorescence showed that Drp1 could protect the mitochondrial ultrastructure in PD-related model mice(p<0.05).Immunoblotting results showed that the expression of Drp1 in mitochondria was significantly down-regulated in mangiferin high-dose group,and the expression of mitophagic proteins PINK1,Parkin,NIX and BNIP3 was up-regulated in mangiferin middle-dose group(p<0.05).Evidence-based medicine research:The results of the meta-analysis showed that the combination of TCM formulas and SSRIs significantly improved the depressive symptoms of Parkinson's disease patients compared with the control group(WMD:-4.920,95%confidence interval:-0.05).5.999,-3.840;p<0.001)and UPDRS ? score(WMD:-1.209,95%confidence interval:-1.561,-0.857;p<0.001).In addition,the results indicated that Chai-Hu-Shu-Gan Powder combined with SSRIs significantly improved the depressive symptoms in Parkinson's disease patients compared to the control group(WMD:-5.390,95%confidence interval:-7.66,-3.11;p<0.001).ConclusionExperimental research:1.In MPTP-PD mouse model,inhibition of Drpl ameliorated the impaired autonomic activity of mice.Furthermore,inhibition of Drpl might protect mitochondrial morphology and ultrastructure by upregulating the expression of PINK1 and Parkin but not BNIP3 and NIX.2.MGF could significantly improve the motor function of PD model mice,decrease the expression of mitochondrial Drpl,and regulate the expression of mitophagic proteins,thereby protecting mitochondrial functions.Evidence-based medicine research:Compared with conventional SSRIs treatment alone,TCM formulas combined with SSRIs treatment could significantly improve depressive symptoms and motor symptoms in PD patients.In addition,we found that Chai-Hu-Shu-Gan Powder combined with SSRIs treatment could significantly improve the depressive symptoms in PD.Thus,the efficacy of TCM formulas combined with SSRIs is superior to that of conventional SSRIs treatment alone,thereby improving the prognosis and quality of life of patients with Parkinson's disease.