Protective effects of Buyinqianzheng Formula on mitochondrial morphology by PINK1/Parkin pathway in SH-SY5Y cells induced by MPP^(+)

Objective:Buyinqianzheng Formula(BYQZF)is clinically employed in traditional Chinese medicine to treat Parkinson's disease(PD)by improving mitochondrial dysfunction.However,the underlying mechanisms by which BYQZF affects mitochondrial morphology remain unknown.Therefore,we observed the effects of BYQZF on mitochondria from the perspective of the PINK1/Parkin pathway.Methods:Cell survival rates were assessed by Cell Counting Kit-8 assay.Expression levels of PINK1 and Parkin mRNA were examined by qRT-PCR.Protein expression levels of PINK1,PINK1-Ser228,Parkin,Parkin-Ser65,Drp1,and Drp1-Ser637 were examined by western blotting.PINK1,Parkin,and MitoTracker?Red CMXRos(MTR)were stained by triple-labeled immunofluorescence,and observed under laser confocal microscopy.Results:Cell survival rate,mitochondrial form factor,mean length and number of mitochondrial network branches,mitochondrial activity,m RNA expression levels of PINK1 and Parkin,and protein expression levels of PINK1,Parkin,and Drp1-Ser637 were reduced after 1-methyl-4-phenylpyridinium(MPP^(+))intervention.In contrast,Pearson's correlation coefficients between PINK1 and Parkin,and between Parkin and MTR,as well as protein expression levels of PINK1-Ser228,Parkin-Ser65,and Drp1 increased significantly after MPP^(+) intervention.Treatment with BYQZF increased cell survival rate,mitochondrial form factor,mean length and number of mitochondrial network branches,mitochondrial activity,m RNA expression levels of PINK1 and Parkin,and expression of PINK1,Parkin,and Drp1-Ser637 proteins.Pearson's correlation coefficients between PINK1 and Parkin,and between Parkin and MTR,as well as protein expression levels of PINK1-Ser228,Parkin-Ser65,and Drp1 decreased after BYQZF treatment.Conclusion:These results demonstrate that BYQZF has a protective effect on mitochondrial molecular mechanisms in the PD cell model,and the mechanism is related to the PINK1/Parkin pathway.

Combination therapy of scalp electro-acupuncture and medication for the treatment of Parkinson's disease: A systematic review and meta-analysis

Objective:To summarize the current clinical evidence related to the therapeutic effects and safety of adjuvant scalp electro-acupuncture (SEA) treatment for Parkinson's disease in China.Methods:Following the PRISMA statement,seven electronic databases were searched to retrieve randomized controlled clinical trials that used SEA combined with medication as the treatment intervention,and medication as the control.RevMan 5.3 was used to analyze outcomes,including the Unified Parkinson's Disease Rating Scale (UPDRS),Webster scale,effectiveness rate,and UPDRS III.Results:Nine randomized controlled trials,with certain methodological flaws and risks of bias,were included that involved 474 participants.SEA combined with medication was more effective than medication alone in overall therapeutic effects,as evidenced by total UPDRS scores (mean difference (MD):7.15,95% confidence interval [CI] 0.24 to 14.07,P =.04),Webster scores (MD:1.60,95% CI 0.20 to 2.99,P =.03),and effectiveness rate (risk ratio:1.35,95% CI 1.19 to 1.54,P <.001).In addition,there was significant improvement in pooled motor function results after adjuvant SEA treatment compared with medication alone (MD:5.75,95% Cl 4.18 to 7.32,P <.001).Conclusion:The combination of SEA and medication may be a promising intervention for patients with Parkinson's disease,especially to improve motor function.However,results were inconclusive,and additional studies with rigorous experimental design and larger sample sizes are needed to verify these results.

Mechanism of mitochondrial protection by the Buyin Qianzheng formula in a Parkin overexpression cell model

Objective:To identify the molecular mechanisms of the effects of the Buyin Qianzheng formula(BYQZF)on the mitochondrial dynamics in a Parkin overexpression Parkinson's disease(PD)cell model.Methods:First,a stable Parkin overexpression cell model was constructed using plasmid transfection.Then,we examined the protective effect of BYQZF on the mitochondrial dysfunction of the Parkin overexpression PD cell model induced by neurotoxin 1-methyl-4-phenylpyridinium ion(MPPþ).The mRNA expression level of Parkin was evaluated using real-time quantitative PCR.The cell survival rate was detected using the Cell Counting Kit-8 assay.We evaluated the cellular adenosine triphosphate(ATP)levels using luciferase assays.A laser scanning confocal microscope was used to observe the mitochondrial morphology,activity,and mitochondrial membrane potential(DJm).Western blot was conducted to evaluate the levels of the fusion proteins mitofusin1,mitofusin2,optic atrophy 1,dynaminrelated protein 1,and mitochondrial fission protein 1.Results:Parkin overexpression attenuated MPPþ-induced mitochondrial damage,increased mitochondrial activity and DJm.BYQZF increased the survival of MPPþ-induced cells that overexpressed Parkin and upregulated the mitochondrial form factor and activity.It also inhibited a decrease in the DJm and ATP levels.These findings suggested that BYQZF protected against MPPþ-induced mitochondrial dysfunction and enhanced the protective effect of Parkin overexpression.Furthermore,the formula upregulated the expression of the fusion proteins mitofusin1,mitofusin2,and optic atrophy 1(closely related to mitochondrial quality remodeling),and reduced the expression of the fission protein dynamicrelated protein 1,as well as mitochondrial fission protein 1.Conclusion:The mechanism by which BYQZF increased the mitochondrial protective effect of Parkin gene overexpression in MPPþ-induced cells may be related to improving mitochondrial function and regulating the balance of mitochondrial division and fusion proteins.

Expression of PNNs and the functional changes of GABA-ergic neurons in the medial prefrontal cortex in a rat model of depression

Objective:To investigate the effects of chronic unpredictable mild stress(CUMS)on perineuronal nets(PNNs)andγ-aminobutyric acid(GABA)-ergic neurons in the medial prefrontal cortex(mPFC)of adult rats.Methods:28 rats were randomly divided into two groups.The model group adopted CUMS to establish a depression model,and the control group did not give any treatment.The density of PNNs and the percentage of PNNs positive(PNNs^(+))and parvalbumin positive(PV^(+))neurons in total PV^(+)neurons in the mPFC were detected by immunofluorescence.The protein expression of main components of PNNs,Aggrecan and Brevican,and GABA main synthase glutamic acid decarboxylase 67(GAD67)in the mPFC were detected by Western blot.Results:The density of PNNs and the percentage of PNNs^(+)and PV^(+)neurons in total PV^(+)neurons in the mPFC in the model group were decreased compared with the control group(P<0.05);The expression levels of PNNs component proteins Aggrecan and Brevican,and GABA main synthase GAD67 were also decreased in the model group(P<0.01).Conclusion:The levels of PNNs and GABA synthase GAD67 in the mPFC of rats were decreased after chronic unpredictable mild stress.

Network pharmacological study of Qingfei Paidu Decoction intervening on cytokine storm mechanism of COVID-19

Objective:To explore the possible mechanism of Qingfei Paibu Decoction in the treatment of COVID-19 from the perspective of cytokine storm by network pharmacology.Methods: The TCMSP and SymMap databases were used to screen out the active components and targets of Qingfei Paibu Decoction;The GeneCards database was used to screen the predicted targets of COVID-19;Two targets were mapped;The STRING database and Cytoscape3.7.2 software were used to construct the network diagram of active drag-ingredient-target and screen out the core components and targets;The GO enrichment analysis and KEGG pathway enrichment analysis were carried out by OmicShare cloud platform and David respectively.Results:A total of 52 potential targets of Qingfei Paibu Decoction for the treatment of COVID-19 were obtained, among which 17 were core targets related to inflammation, mainly including cytokines such as IL, IFN, TNF and chemokines. And 26 core components were obtained, including quercetin, luteolin, kaempferol, naringenin, and baicalein;The GO enrichment results showed 224 biological processes, 15 molecular functions and 33 cell components related to inflammation;There were 5 inflammatory signaling pathways in KEGG enrichment results, including TNF signaling pathway, Nod-like receptor signaling pathway, Toll-like receptor signaling pathway, MAPK signaling pathway and cytokine receptor interaction. Conclusion: Qingfei Paibu Decoction can inhibit cytokine storms by acting on multiple targets and pathways with multiple components and thus treat COVID-19.

Effects of stress of different duration on depression-like behavior and expression of CB1 and GluA1 in medial prefrontal cortex of rats

Objective:To explore the changes of cannabinoid receptor(CB1)and AMPA receptor subunit GluA1 in the medial prefrontal cortex(mPFC)of depression model rats induced by chronic unpredictable mild stress(CUMS).Methods:30 rats were randomly divided into three groups.The three-week model group was given a three-week CUMS model,the four-week model group was given a four-week CUMS model,and the control group was given no treatment.The behavior of rats were detected in each group,and the expression of CB1 and GluA1 protein in synaptosomes in mPFC brain region was detected by Western blot.Results:Compared with the control group,the surcose preference decreased,the feeding latency increased in the novel inhibition feeding test,and the immobility time increased in the forced swimming test in the four-week model group.However,after three weeks of CUMS,there was no obvious change in the behavior of rats compared with the control group,suggesting that four-week model of CUMS was successful.CUMS can reduce the expression level of CB1 and GluA1 protein in synaptosomes of mPFC brain area in four-week model group(P<0.05),but there was no significant difference between three-week model group and control group.Conclusion:Four-week CUMS was more likely to lead to depressive-like behavior in rats,which may be closely related to the expression of CB1 and GluA1 in mPFC.