Objective The neuroprotective effect of erythropoietin （EPO） against 1-methyl-4-phenylpyridinium （MPP^＋）- induced oxidative stress in cultured PC12 cells, as well as the underlying mechanism, were investigated. M...Objective The neuroprotective effect of erythropoietin （EPO） against 1-methyl-4-phenylpyridinium （MPP^＋）- induced oxidative stress in cultured PC12 cells, as well as the underlying mechanism, were investigated. Methods PC12 ceils impaired by MPP^＋ were used as the cell model of Parkinson＇s disease. Methyl thiazolyl tetrazolium （MTT） was used to assay the viability of the PC12 cells exposed to gradient concentrations of EPO, and the terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） assay was used to analyze the apoptosis ratio of PC 12 cells. The expression of Bcl-2 and Bax in PC 12 cells were examined by Western blot, and the reactive oxygen species （ROS）, the mitochondrial transmembrane potential and the activity of caspase-3 in each group were detected by spectrofluorometer. Results Treatment of PC12 cells with MPP^＋ caused the loss of cell viability, which may be associated with the elevation in apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. It was also shown that MPP＋ significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, EPO significantly reversed these responses and had the maximum protective effect at 1 U/mL. Conclusion The inhibitive effect of EPO on the MPP^＋ -induced cytotoxicity may be ascribed to its anti-oxidative property and anti-apoptotic activity, and EPO may provide a useful therapeutic strategy for treatment of neurodegenerative diseases such as Parkinson＇s disease.展开更多
BACKGROUND： Insulin receptor （IR） expression in the substantia nigra of patients with Parkinson disease （PD） is not only significantly lower than that in the substantia nigra of normal persons of the same age, bu...BACKGROUND： Insulin receptor （IR） expression in the substantia nigra of patients with Parkinson disease （PD） is not only significantly lower than that in the substantia nigra of normal persons of the same age, but also significantly lower than that in other regions in brain of himself/herself. It suggests that the abnormal effect of insulin receptor-mediated insulin, as a neurotrophic factor, is very possibly related to the loss of dopaminergic neurons in the substantia nigra and striatum in patients with Parkinson disease. OBJECTIVE ： TO observe the interventional effect of insulin on 1-methyl-4-phenylpyridinium ion （MPP^＋）-induced apoptosis of PC12. DESIGN： Controlled observation SETTINGS： Department of Neurology, Beijing China-Japan Friendship Hospital; Department of Neurology Huashan Hospital Affiliated to Fudan University. MATERIALS： PC12 cells were provided by the Cell Bank, Shanghai Institute of Cell Biology, Chinese Academy of Science. MPP^＋, MTT, HOECHST 33258 （Invitrogen Life Technologies）, reverse transcription-polymerase chain reaction （RT-PCR） reagent （Takara Shuzo Co., Ltd.）, flow cytometer （Bacton Dickionson, San Jose, CA）, enzyme labelling instrument （Bio-Tek, Winooski, VT） and PCR circulation instrument （Takara Shuzo Co., Ltd） were used in this study. METHODS ： This study was carried out in the Department of Neurology, Huashan Hospital Affiliated to Fudan University during June 2003 to August 2004. （1） Cell culture and experimental grouping： PC12 cells were cultured according to the method from Peng et al, then were randomized into 3 groups; blank control group, MPP^＋ group and insulin group. （2） Detection of relative survival rate of cells： The relative survival rate of cells at different MPP^＋ final concentrations （0, 50, 100, 200, 300, 1 000 μmol/L） and at different culture time （0, 4, 8, 12, 18, 24 hours） in the 300 Fmol/L MPP^＋ group and different concentrations of insulin （0, 15, 50, 100 nmol/L） in the insulin group was detected with MTT method according to the method from Hansen et al. （3） Observation of cell apoptosis： After stained by HOECHST 33258, the apoptotic cells were observed under the fluorescence miscroscope with the method from Chen et al. （4） Dection of apoptotic rate of cells： Apoptotic rate of cells was detected with flow cytometry according to the method from Zhang et al. （5） The expression of tyrosine hydroxylase （TH） mRNA in PC12 cells was detected with RT-PCR methods according to the modified method from Peng et al. MAIN OUTCOME MEASURES ： Comparison of relative survival rate, apoptosis rate, the expression of IR mRNA and TH mRNA and cell apoptosis. RESULTS： （1） After 12-hour incubation of 100, 200, 300 and 1 000 μmol/L MPP^＋, the relative survival rate of PC12 cells was （72.88±2.91）%, （60.64±0.81）%, （54.56±0.76）% and （16.89±2.83）%, respectively, which was significantly lower than that of blank control group （100%, P 〈 0.05）; After 12, 18 and 24-hour incubation, the relative survival rate of PC12 cells was （54.56±0.76）%, （42.43±0.16）% and （23.56±0.17）% respectively, which was significantly lower than that of blank control group （100%, P〈 0.05）; When 15, 50 and 100 nmol/L insulin was pre-added to cells, the relative survival rate was （70.10±0.16）%, （78.01 ±2.43）% and （83.55±1.43）%, respectively, which was significantly higher than MPP^＋ alone [（54.56±0.76）%, P 〈 0.05]. （2） Appototic bodies were rarely seen in the blank control group, but densely gathered in the MPP^＋ group and were significantly decreased in the insulin group. （3） Apoptosis rate of PC12 cells in the MPP^＋ group was significantly higher than that in the blank control group [（36.56±0.89）% vs. （2.34±0.23）%, P〈 0.05], and that in the 15, 50, 100 nmol/L insulin group [（30.01±0.04）%, （24.23±0.37）%, （20.01 ±1.01）%, respectivelyl was significantly lower than that in MPP^＋ group （P 〈 0.05）. （4） The TH mRNA expression in PC12 cells in MPP^＋ group was significantly lower than that in blank control group; The expression of TH mRNA in insulin group was gradually increased in an insulin dose-dependent manner. There were no significant changes in the expression of IR mRNA under different experimental conditions. CONCLUSION： Insulin can resist MPP^＋-induced apoptosis of PC12 cells, lessen the damage of PC12 cells, but does not change the gene expression of target cell insulin receptor.展开更多
1-methyl-4-phenylpyridinium ion （MPP^＋） induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The p...1-methyl-4-phenylpyridinium ion （MPP^＋） induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The present study investigated the regulatory effects of nerve growth factor （Akt activator） and lithium chloride （glycogen synthase kinase-3β inhibitor） on the endoplasmic reticulum stress signaling pathway. The results revealed that MPP＋ induced expression of Bip and C/EBP homologous protein. The upregulation of Bip and C/EBP homologous protein, as well as the decreased pro-caspase-12 level induced by MPP^＋ were inhibited by pretreatment of the nerve growth factor or lithium chloride. These results suggest that the phosphatidylinositol 3 kinase-Aktglycogen synthase kinase-3β pathway is involved in MPP-induced endoplasmic reticulum stress.展开更多
Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion （MPP＋）-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-（4,5-Dimethylthia...Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion （MPP＋）-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay, respectively. The release of lactate dehydrogenase, superoxide dismutase activity and levels of malondialdehyde were determined by UV spectrophotometry. The changes in mitochondrial membrane potential and the intracellular concentration of calcium were determined by flow cytometry, and the activity of caspase-3 was monitored by western blot. According to cell viability and apoptosis studies, MPP＋-induced apoptosis in PC12 cells was inhibited in the presence of 10 tJg/mL of Eleutheroside B Our results indicate that the neuroprotective effect of Eleutheroside B, following MPP＋-induced apoptosis in PC12 cells, involves increasing the anti-oxidative stress capacity of cells, maintaining the high-energy state of mitochondrial membrane potential, reducing intracellular calcium concentration and inhibiting caspase-3 activity.展开更多
Microglia-mediated neuroinflammation is considered a pathological feature of Parkinson's disease.Triggering receptor expressed on myeloid cell-1(TREM-1)can amplify the inherent immune response,and crucially,regula...Microglia-mediated neuroinflammation is considered a pathological feature of Parkinson's disease.Triggering receptor expressed on myeloid cell-1(TREM-1)can amplify the inherent immune response,and crucially,regulate inflammation.In this study,we found marked elevation of serum soluble TREM-1 in patients with Parkinson's disease that positively correlated with Parkinson's disease severity and dyskinesia.In a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson's disease,we found that microglial TREM-1 expression also increased in the substantia nigra.Further,TREM-1 knockout alleviated dyskinesia in a mouse model of Parkinson's disease and reduced dopaminergic neuronal injury.Meanwhile,TREM-1 knockout attenuated the neuroinflammatory response,dopaminergic neuronal injury,and neutrophil migration.Next,we established an in vitro 1-methyl-4-phenyl-pyridine-induced BV2 microglia model of Parkinson's disease and treated the cells with the TREM-1 inhibitory peptide LP17.We found that LP17 treatment reduced apoptosis of dopaminergic neurons and neutrophil migration.Moreover,inhibition of neutrophil TREM-1 activation diminished dopaminergic neuronal apoptosis induced by lipopolysaccharide.TREM-1 can activate the downstream CARD9/NF-κB proinflammatory pathway via interaction with SYK.These findings suggest that TREM-1 may play a key role in mediating the damage to dopaminergic neurons in Parkinson's disease by regulating the interaction between microglia and peripheral neutrophils.展开更多
The positive effect of levodopa in the treatment of Parkinson’s disease,although it is limited in time and has severe side effects,has encouraged the scientific community to look for new drugs that can stop the neuro...The positive effect of levodopa in the treatment of Parkinson’s disease,although it is limited in time and has severe side effects,has encouraged the scientific community to look for new drugs that can stop the neurodegenerative process or even regenerate the neuromelanin-containing dopaminergic nigrostriatal neurons.Successful preclinical studies with coenzyme Q10,mitoquinone,isradipine,nilotinib,TCH346,neurturin,zonisamide,deferiprone,prasinezumab,and cinpanemab prompted clinical trials.However,these failed and after more than 50 years levodopa continues to be the key drug in the treatment of the disease,despite its severe side effects after 4–6 years of chronic treatment.The lack of translated successful results obtained in preclinical investigations based on the use of neurotoxins that do not exist in the human body as new drugs for Parkinson’s disease treatment is a big problem.In our opinion,the cause of these failures lies in the experimental animal models involving neurotoxins that do not exist in the human body,such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and 6-hydroxydopamine,that induce a very fast,massive and expansive neurodegenerative process,which contrasts with the extremely slow one of neuromelanin-containing dopaminergic neurons.The exceedingly slow progress of the neurodegenerative process of the nigrostriatal neurons in idiopathic Parkinson’s patients is due to(i)a degenerative model in which the neurotoxic effect of an endogenous neurotoxin affects a single neuron,(ii)a neurotoxic event that is not expansive and(iii)the fact that the neurotoxin that triggers the neurodegenerative process is produced inside the neuromelanin-containing dopaminergic neurons.The endogenous neurotoxin that fits this degenerative model involving one single neuron at a time is aminochrome,since it(i)is generated within neuromelanin-containing dopaminergic neurons,(ii)does not cause an expansive neurotoxic effect and(iii)triggers all the mechanisms involved in the neurodegenerative process of the nigrostriatal neurons in idiopathic Parkinson’s disease.In conclusion,based on the hypothesis that the neurodegenerative process of idiopathic Parkinson’s disease corresponds to a single-neuron neurodegeneration model,we must search for molecules that increase the expression of the neuroprotective enzymes DT-diaphorase and glutathione transferase M2-2.It has been observed that the activation of the Kelch-like ECH-associated protein 1/nuclear factor(erythroid-derived 2)-like 2 pathway is associated with the transcriptional activation of the DT-diaphorase and glutathione transferase genes.展开更多
Objective:Mitophagy is known to contribute towards progression of Parkinson’s disease.Korean red ginseng(KRG)is a widely used medicinal herb in East Asia,and recent studies have reported that KRG prevents 1-methyl-4-...Objective:Mitophagy is known to contribute towards progression of Parkinson’s disease.Korean red ginseng(KRG)is a widely used medicinal herb in East Asia,and recent studies have reported that KRG prevents 1-methyl-4-phenylpyridinium ion(MPP^(+))-induced cell death.This study was undertaken to investigate whether KRG suppresses MPP^(+)-induced apoptosis and mitophagy.Methods:SH-SY5 Y cells were incubated with KRG for 24 h,and subsequently exposed to MPP^(+).The MPP^(+)-induced cell death was confirmed with the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay,and the terminal deoxynucleotidyl transferase-mediated d UTP nick end-labeling assay.Changes in the structure and function of mitochondria were confirmed using mitotracker,Mito SOX red mitochondrial superoxide indicator,parkin,and phosphatase and tensin homolog deleted on chromosome ten-induced putative kinase 1(PINK1)immunofluorescent staining.Western blotting was performed to evaluate the expression of apoptosis-related factors in whole cells,including Bax,Bcl-2 and cleaved caspase-3,and mitophagy-related factors in the mitochondrial fraction,including cytochrome c,parkin,PINK1,translocase of the outer membrane 20(TOM20),p62 and Beclin 1.Results:MPP^(+)induced cell death by cytochrome c release and caspase-3 activation;however,this effect was suppressed by KRG’s regulation of the expressions of Bcl-2 and Bax.Moreover,MPP^(+)exposure increased the mitochondrial expressions of parkin,PINK1,Beclin 1 and p62,and decreased TOM20,cytochrome c and Bcl-2 expressions.These MPP^(+)-induced changes in the mitochondrial fraction were attenuated by treatment with KRG.Conclusion:KRG effectively prevents MPP^(+)-induced SH-SY5 Y cell death by regulating cytochrome c release from mitochondria and PINK1/parkin-mediated mitophagy,through regulation of the Bcl-2 family.展开更多
Mitochondrial superoxide overproduction is believed to be responsible for the neurotoxicity associated with neurodegeneration.Mitochondria-targeted antioxidants,such as MitoQ,have emerged as potentially effective anti...Mitochondrial superoxide overproduction is believed to be responsible for the neurotoxicity associated with neurodegeneration.Mitochondria-targeted antioxidants,such as MitoQ,have emerged as potentially effective antioxidant therapies.Methionine sulfoxide reductase A(MsrA)is a key mitochondrial-localized endogenous antioxidative enzyme and it can scavenge oxidizing species by catalyzing the methionine(Met)-centered redox cycle(MCRC).In this study,we observed that the natural L-Met acted as a good scavenger for antimycin A-induced mitochondrial superoxide overproduction in PC12 cells.This antioxidation was largely dependent on the Met oxidase activity of MsrA.S-methyl-L-cysteine(SMLC),a natural analogue of Met that is abundantly found in garlic and cabbage,could activate the Met oxidase activity of MsrA to scavenge free radicals.Furthermore,SMLC protected against antimycin A-induced mitochondrial membrane depolarization and alleviated 1-methyl-4-phenylpyridinium(MPP+)-induced neurotoxicity.Thus,our data highlighted the possibility for SMLC supplement in the detoxication of mitochondrial damage by activating the Met oxidase activity of MsrA.展开更多
In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1（SIAH-1） in PC12 cells. 1-Methyl-4-phenylpyridinium（MPP＋） treatment increased α-synuclein, E1 and S...In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1（SIAH-1） in PC12 cells. 1-Methyl-4-phenylpyridinium（MPP＋） treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression. Treatment with an SIAH-1 antibody decreased m RNA expression levels of α-synuclein, light chain 3 and SIAH-1, but increased E1 m RNA expression. It also increased cell viability. Combined treatment with MPP＋ and rapamycin reduced SIAH-1 and α-synuclein levels. Treatment with SIAH-1 antibody alone diminished α-synuclein immunoreactivity in PC12 cells, and reduced the colocalization of α-synuclein and light chain 3. These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway. Consequently, SIAH-1 may be a potential new therapeutic target for Parkinson’s disease.展开更多
The transient receptor potential melastatin 2 is a calcium-permeable cation channel member of the TRP family. Also known as an oxidative stress-activated channel, the transient receptor potential melastatin 2 gating m...The transient receptor potential melastatin 2 is a calcium-permeable cation channel member of the TRP family. Also known as an oxidative stress-activated channel, the transient receptor potential melastatin 2 gating mechanism is dependent on reactive oxygen species. In pathological conditions, transient receptor potential melastatin 2 is overactivated, leading to a Ca~(2+) influx that alters cell homeostasis and promotes cell death. The role of transient receptor potential melastatin 2 in neurodegenerative diseases, including Alzheimer's disease and ischemia, has already been described and reviewed. However, data on transient receptor potential melastatin 2 involvement in Parkinson's disease pathology has emerged only in recent years and the issue lacks review studies that focus specifically on this topic. The present review aims to elucidate the role of the transient receptor potential melastatin 2 channel in Parkinson's disease by reviewing, summarizing, and discussing the in vitro, in vivo, and human studies published until August 2022. Here we describe fourteen studies that evaluated the transient receptor potential melastatin 2 channel in Parkinson's disease. The Parkinson's disease model used, transient receptor potential melastatin 2 antagonist and genetic approaches, and the main outcomes reported were discussed. The studies described transient receptor potential melastatin 2 activation and enhanced expression in different Parkinson's disease models. They also evidenced protective and restorative effects when using transient receptor potential melastatin 2 antagonists, knockout, or silencing. This review provides a literature overview and suggests where there is a need for more research. As a perspective point, this review shows evidence that supports transient receptor potential melastatin 2 as a pharmacological target for Parkinson's disease in the future.展开更多
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 ...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.展开更多
Objective:To investigate the neuroprotective effects of Fructus Mume total flavone(FMF)against cell apoptosis and mitochondrial injury induced by 1-methyl-4-phenylpyridinium(MPP^(+))in human neuroblastoma(SH-SY5Y)cell...Objective:To investigate the neuroprotective effects of Fructus Mume total flavone(FMF)against cell apoptosis and mitochondrial injury induced by 1-methyl-4-phenylpyridinium(MPP^(+))in human neuroblastoma(SH-SY5Y)cells and explore its molecular mechanisms.Methods:MPP^(+) induced SH-SY5Y cells injury model were established in vitro cell culture,the cells were divided into 5 groups:normal control group,model group(250μmol·L^(-1) MPP^(+)),FMF low-and middle-and high-dose experimental group(10,50,100μmol·L^(-1) FMF).After 72 h administration,4’,6-diamidino-2-phenylindole(DAPI)staining was used to observe the effects of different concentrations of FMF on the morphologic changes of apoptotic cells,the ratio of cell apoptosis was measured by Annexin-FITC/PI double staining.The mitochondrial membrane electro-bit were detected by flow cytometry(FCM).The expression of Bcl-2,Bax and Caspase-3 were detected by Western blot.Results:The results of DAPI staining showed that the injury SH-SY5Y cells induced by MPP+were densely condensed,the nucleus showed nuclear shrinkage,showing an apoptotic characteristic morphology;after 72h of FMF action,the apoptotic morphology of the cells showed different degrees of improvement,and the apoptotic number of SH-SY5Y cells also decreased.Compared with that in the normal control group,the apoptotic rate and of mitochondrial membrane electrobit of SH-SY5Y cells in the model group increased significantly(P<0.01),the expression of Bax and Caspase-3 proteins increased significantly(P<0.01),Bcl-2 protein and the ratio of Bcl-2/Bax decreased significantly(P<0.01).Compared with that in the model group,the apoptotic rate and mitochondrial membrane electro-bit of SH-SY5Y cells in FMF groups(10,50,100μmol·L^(-1))were significantly lower,while Bax and Caspase-3 proteins were significantly lower(P<0.01),and Bcl-2 protein and the ratio of Bcl-2/Bax were significantly higher,with statistically significant difference in FMF middle-and high-dose experimental groups(P<0.01).The results indicated that FMF can decrease the experession level of Bax and Caspase-3 and increase the ratio of Bcl-2/Bax,inhibit MPP+induced apoptosis.Conclusion:FMF improves the damage of SH-SY5Y cells induced by MPP+,and plays a neuroprotective effect by regulating the expressions of related proteins in mitochondrial apoptosis pathway.展开更多
Echinacoside （ECH） is protective in a mouse model of Parkinson＇ s disease （PD） induced by 1-methyl-4- phenylpyridinium ion （MPP＋）. To investigate the mechanisms involved, SH-SYSY neuroblastoma ceils were treat...Echinacoside （ECH） is protective in a mouse model of Parkinson＇ s disease （PD） induced by 1-methyl-4- phenylpyridinium ion （MPP＋）. To investigate the mechanisms involved, SH-SYSY neuroblastoma ceils were treated with MPP＋ or a combination of MPP＋ and ECH, and the expression of ATF3 （activating transcription factor 3）, CHOP （C/EBP-homologous protein）, SCNA （synuclein alpha）, and GDNF （glial cell line-derived neurotrophic factor） was assessed. The results showed that ECH significantly improved cell survival by inhibiting the generation of MPP＋-induced reactive oxygen species （ROS）. In addition, ECH suppressed the ROS and MPP＋- induced expression of apoptotic genes （ATF3, CHOP, and SCNA）. ECH markedly decreased the MPP＋-induced cas- pase-3 activity in a dose-dependent manner. ATF3- knockdown also decreased the CHOP and cleaved caspase- 3 levels and inhibited the apoptosis induced by MPP＋. Interestingly, ECH partially restored the GDNF expression that was down-regulated by MPP＋. ECH also improved dopaminergic neuron survival during MPP＋ treatment and protected these neurons against the apoptosis induced by MPTP. Taken together, these data suggest that the ROS/ ATF3/CHOP pathway plays a critical role in mechanisms by which ECH protects against MPP＋-induced apoptosis in PD.展开更多
Objective:The objective of this study was to investigate whether 70%aqueous ethanol extract of San-Jia-Fu-Mai decoction extract(SJFMDE)could protect against 1-methyl-4-phenylpyridinium(MPP+)-induced oxidative stress i...Objective:The objective of this study was to investigate whether 70%aqueous ethanol extract of San-Jia-Fu-Mai decoction extract(SJFMDE)could protect against 1-methyl-4-phenylpyridinium(MPP+)-induced oxidative stress in PC12 cells and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced motor function deficits in mice.Materials and Methods:The cell viability,the levels of intracellular reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)in the MPP+-treated PC12 cells were measured.Motor function deficits and dopamine(DA)level in the brain striatum and tyrosine hydroxylase(TH)-positive cells in substantia nigra pars compacta(SNc)of the MPTP-treated mice were determined.Results:The results showed that SJFMDE could reduce cell death and the levels of ROS and MDA while increase the level of GSH in the MPP+-treated PC12 cells.In addition,in vivo studies showed that oral administration of SJFMDE(3,6,and 12 g/kg)significantly improved the motor function deficits induced by MPTP and enhanced the DA level in the striatum and TH-positive neuronal cells in SNc of the MPTP-treated mice.Conclusions:Our results revealed that SJFMDE possessed neuroprotective effects against neurotoxicity induced by MPP+and motor function deficits induced by MPTP via suppressing oxidative stress and increasing the levels of DA and TH,indicating that SJFMDE might be a promising Chinese medicine formula for the treatment of Parkinson’s disease.展开更多
文摘Objective The neuroprotective effect of erythropoietin （EPO） against 1-methyl-4-phenylpyridinium （MPP^＋）- induced oxidative stress in cultured PC12 cells, as well as the underlying mechanism, were investigated. Methods PC12 ceils impaired by MPP^＋ were used as the cell model of Parkinson＇s disease. Methyl thiazolyl tetrazolium （MTT） was used to assay the viability of the PC12 cells exposed to gradient concentrations of EPO, and the terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） assay was used to analyze the apoptosis ratio of PC 12 cells. The expression of Bcl-2 and Bax in PC 12 cells were examined by Western blot, and the reactive oxygen species （ROS）, the mitochondrial transmembrane potential and the activity of caspase-3 in each group were detected by spectrofluorometer. Results Treatment of PC12 cells with MPP^＋ caused the loss of cell viability, which may be associated with the elevation in apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. It was also shown that MPP＋ significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, EPO significantly reversed these responses and had the maximum protective effect at 1 U/mL. Conclusion The inhibitive effect of EPO on the MPP^＋ -induced cytotoxicity may be ascribed to its anti-oxidative property and anti-apoptotic activity, and EPO may provide a useful therapeutic strategy for treatment of neurodegenerative diseases such as Parkinson＇s disease.
文摘BACKGROUND： Insulin receptor （IR） expression in the substantia nigra of patients with Parkinson disease （PD） is not only significantly lower than that in the substantia nigra of normal persons of the same age, but also significantly lower than that in other regions in brain of himself/herself. It suggests that the abnormal effect of insulin receptor-mediated insulin, as a neurotrophic factor, is very possibly related to the loss of dopaminergic neurons in the substantia nigra and striatum in patients with Parkinson disease. OBJECTIVE ： TO observe the interventional effect of insulin on 1-methyl-4-phenylpyridinium ion （MPP^＋）-induced apoptosis of PC12. DESIGN： Controlled observation SETTINGS： Department of Neurology, Beijing China-Japan Friendship Hospital; Department of Neurology Huashan Hospital Affiliated to Fudan University. MATERIALS： PC12 cells were provided by the Cell Bank, Shanghai Institute of Cell Biology, Chinese Academy of Science. MPP^＋, MTT, HOECHST 33258 （Invitrogen Life Technologies）, reverse transcription-polymerase chain reaction （RT-PCR） reagent （Takara Shuzo Co., Ltd.）, flow cytometer （Bacton Dickionson, San Jose, CA）, enzyme labelling instrument （Bio-Tek, Winooski, VT） and PCR circulation instrument （Takara Shuzo Co., Ltd） were used in this study. METHODS ： This study was carried out in the Department of Neurology, Huashan Hospital Affiliated to Fudan University during June 2003 to August 2004. （1） Cell culture and experimental grouping： PC12 cells were cultured according to the method from Peng et al, then were randomized into 3 groups; blank control group, MPP^＋ group and insulin group. （2） Detection of relative survival rate of cells： The relative survival rate of cells at different MPP^＋ final concentrations （0, 50, 100, 200, 300, 1 000 μmol/L） and at different culture time （0, 4, 8, 12, 18, 24 hours） in the 300 Fmol/L MPP^＋ group and different concentrations of insulin （0, 15, 50, 100 nmol/L） in the insulin group was detected with MTT method according to the method from Hansen et al. （3） Observation of cell apoptosis： After stained by HOECHST 33258, the apoptotic cells were observed under the fluorescence miscroscope with the method from Chen et al. （4） Dection of apoptotic rate of cells： Apoptotic rate of cells was detected with flow cytometry according to the method from Zhang et al. （5） The expression of tyrosine hydroxylase （TH） mRNA in PC12 cells was detected with RT-PCR methods according to the modified method from Peng et al. MAIN OUTCOME MEASURES ： Comparison of relative survival rate, apoptosis rate, the expression of IR mRNA and TH mRNA and cell apoptosis. RESULTS： （1） After 12-hour incubation of 100, 200, 300 and 1 000 μmol/L MPP^＋, the relative survival rate of PC12 cells was （72.88±2.91）%, （60.64±0.81）%, （54.56±0.76）% and （16.89±2.83）%, respectively, which was significantly lower than that of blank control group （100%, P 〈 0.05）; After 12, 18 and 24-hour incubation, the relative survival rate of PC12 cells was （54.56±0.76）%, （42.43±0.16）% and （23.56±0.17）% respectively, which was significantly lower than that of blank control group （100%, P〈 0.05）; When 15, 50 and 100 nmol/L insulin was pre-added to cells, the relative survival rate was （70.10±0.16）%, （78.01 ±2.43）% and （83.55±1.43）%, respectively, which was significantly higher than MPP^＋ alone [（54.56±0.76）%, P 〈 0.05]. （2） Appototic bodies were rarely seen in the blank control group, but densely gathered in the MPP^＋ group and were significantly decreased in the insulin group. （3） Apoptosis rate of PC12 cells in the MPP^＋ group was significantly higher than that in the blank control group [（36.56±0.89）% vs. （2.34±0.23）%, P〈 0.05], and that in the 15, 50, 100 nmol/L insulin group [（30.01±0.04）%, （24.23±0.37）%, （20.01 ±1.01）%, respectivelyl was significantly lower than that in MPP^＋ group （P 〈 0.05）. （4） The TH mRNA expression in PC12 cells in MPP^＋ group was significantly lower than that in blank control group; The expression of TH mRNA in insulin group was gradually increased in an insulin dose-dependent manner. There were no significant changes in the expression of IR mRNA under different experimental conditions. CONCLUSION： Insulin can resist MPP^＋-induced apoptosis of PC12 cells, lessen the damage of PC12 cells, but does not change the gene expression of target cell insulin receptor.
基金the National Natural Science Foundation of China, No. 30860085a grant from the Candidates of Young and Middle-Aged Academic Leaders of Yunnan Province, No. 2006PY01-07the Natural Science Foundation of Yunnan Province, No. 2007C177M
文摘1-methyl-4-phenylpyridinium ion （MPP^＋） induces endoplasmic reticulum stress and activates caspase-12 in PC12 cells, leading to neuronal apoptosis. However, the underlying molecular mechanism remains unknown. The present study investigated the regulatory effects of nerve growth factor （Akt activator） and lithium chloride （glycogen synthase kinase-3β inhibitor） on the endoplasmic reticulum stress signaling pathway. The results revealed that MPP＋ induced expression of Bip and C/EBP homologous protein. The upregulation of Bip and C/EBP homologous protein, as well as the decreased pro-caspase-12 level induced by MPP^＋ were inhibited by pretreatment of the nerve growth factor or lithium chloride. These results suggest that the phosphatidylinositol 3 kinase-Aktglycogen synthase kinase-3β pathway is involved in MPP-induced endoplasmic reticulum stress.
基金the Major Projects of National Science and Technology, No.2009ZX09103-329the National Natural Science Foundation for Distinguished Young Scholars of China, No.30901974+1 种基金Outstanding Youth Science Fund Program of Heilongjiang Province, No.JC200705"Spring Sunshine" Plan of Ministry of Education, No.2006
文摘Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion （MPP＋）-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay, respectively. The release of lactate dehydrogenase, superoxide dismutase activity and levels of malondialdehyde were determined by UV spectrophotometry. The changes in mitochondrial membrane potential and the intracellular concentration of calcium were determined by flow cytometry, and the activity of caspase-3 was monitored by western blot. According to cell viability and apoptosis studies, MPP＋-induced apoptosis in PC12 cells was inhibited in the presence of 10 tJg/mL of Eleutheroside B Our results indicate that the neuroprotective effect of Eleutheroside B, following MPP＋-induced apoptosis in PC12 cells, involves increasing the anti-oxidative stress capacity of cells, maintaining the high-energy state of mitochondrial membrane potential, reducing intracellular calcium concentration and inhibiting caspase-3 activity.
基金supported by the National Natural Science Foundation of China,Nos.82271257(to YZ)and 82071228(to YZ)Qing Lan Project(to YZ)+1 种基金Open Competition Grant of Xuzhou Medical University(to YZ)Postgraduate Research&Practice Innovation Program of Jiangsu Province,No.KYCX21_2705(to TS)。
文摘Microglia-mediated neuroinflammation is considered a pathological feature of Parkinson's disease.Triggering receptor expressed on myeloid cell-1(TREM-1)can amplify the inherent immune response,and crucially,regulate inflammation.In this study,we found marked elevation of serum soluble TREM-1 in patients with Parkinson's disease that positively correlated with Parkinson's disease severity and dyskinesia.In a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson's disease,we found that microglial TREM-1 expression also increased in the substantia nigra.Further,TREM-1 knockout alleviated dyskinesia in a mouse model of Parkinson's disease and reduced dopaminergic neuronal injury.Meanwhile,TREM-1 knockout attenuated the neuroinflammatory response,dopaminergic neuronal injury,and neutrophil migration.Next,we established an in vitro 1-methyl-4-phenyl-pyridine-induced BV2 microglia model of Parkinson's disease and treated the cells with the TREM-1 inhibitory peptide LP17.We found that LP17 treatment reduced apoptosis of dopaminergic neurons and neutrophil migration.Moreover,inhibition of neutrophil TREM-1 activation diminished dopaminergic neuronal apoptosis induced by lipopolysaccharide.TREM-1 can activate the downstream CARD9/NF-κB proinflammatory pathway via interaction with SYK.These findings suggest that TREM-1 may play a key role in mediating the damage to dopaminergic neurons in Parkinson's disease by regulating the interaction between microglia and peripheral neutrophils.
文摘The positive effect of levodopa in the treatment of Parkinson’s disease,although it is limited in time and has severe side effects,has encouraged the scientific community to look for new drugs that can stop the neurodegenerative process or even regenerate the neuromelanin-containing dopaminergic nigrostriatal neurons.Successful preclinical studies with coenzyme Q10,mitoquinone,isradipine,nilotinib,TCH346,neurturin,zonisamide,deferiprone,prasinezumab,and cinpanemab prompted clinical trials.However,these failed and after more than 50 years levodopa continues to be the key drug in the treatment of the disease,despite its severe side effects after 4–6 years of chronic treatment.The lack of translated successful results obtained in preclinical investigations based on the use of neurotoxins that do not exist in the human body as new drugs for Parkinson’s disease treatment is a big problem.In our opinion,the cause of these failures lies in the experimental animal models involving neurotoxins that do not exist in the human body,such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and 6-hydroxydopamine,that induce a very fast,massive and expansive neurodegenerative process,which contrasts with the extremely slow one of neuromelanin-containing dopaminergic neurons.The exceedingly slow progress of the neurodegenerative process of the nigrostriatal neurons in idiopathic Parkinson’s patients is due to(i)a degenerative model in which the neurotoxic effect of an endogenous neurotoxin affects a single neuron,(ii)a neurotoxic event that is not expansive and(iii)the fact that the neurotoxin that triggers the neurodegenerative process is produced inside the neuromelanin-containing dopaminergic neurons.The endogenous neurotoxin that fits this degenerative model involving one single neuron at a time is aminochrome,since it(i)is generated within neuromelanin-containing dopaminergic neurons,(ii)does not cause an expansive neurotoxic effect and(iii)triggers all the mechanisms involved in the neurodegenerative process of the nigrostriatal neurons in idiopathic Parkinson’s disease.In conclusion,based on the hypothesis that the neurodegenerative process of idiopathic Parkinson’s disease corresponds to a single-neuron neurodegeneration model,we must search for molecules that increase the expression of the neuroprotective enzymes DT-diaphorase and glutathione transferase M2-2.It has been observed that the activation of the Kelch-like ECH-associated protein 1/nuclear factor(erythroid-derived 2)-like 2 pathway is associated with the transcriptional activation of the DT-diaphorase and glutathione transferase genes.
基金financially supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Korea government(MSIT)(No.NRF-2019R1A2C1085130)。
文摘Objective:Mitophagy is known to contribute towards progression of Parkinson’s disease.Korean red ginseng(KRG)is a widely used medicinal herb in East Asia,and recent studies have reported that KRG prevents 1-methyl-4-phenylpyridinium ion(MPP^(+))-induced cell death.This study was undertaken to investigate whether KRG suppresses MPP^(+)-induced apoptosis and mitophagy.Methods:SH-SY5 Y cells were incubated with KRG for 24 h,and subsequently exposed to MPP^(+).The MPP^(+)-induced cell death was confirmed with the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay,and the terminal deoxynucleotidyl transferase-mediated d UTP nick end-labeling assay.Changes in the structure and function of mitochondria were confirmed using mitotracker,Mito SOX red mitochondrial superoxide indicator,parkin,and phosphatase and tensin homolog deleted on chromosome ten-induced putative kinase 1(PINK1)immunofluorescent staining.Western blotting was performed to evaluate the expression of apoptosis-related factors in whole cells,including Bax,Bcl-2 and cleaved caspase-3,and mitophagy-related factors in the mitochondrial fraction,including cytochrome c,parkin,PINK1,translocase of the outer membrane 20(TOM20),p62 and Beclin 1.Results:MPP^(+)induced cell death by cytochrome c release and caspase-3 activation;however,this effect was suppressed by KRG’s regulation of the expressions of Bcl-2 and Bax.Moreover,MPP^(+)exposure increased the mitochondrial expressions of parkin,PINK1,Beclin 1 and p62,and decreased TOM20,cytochrome c and Bcl-2 expressions.These MPP^(+)-induced changes in the mitochondrial fraction were attenuated by treatment with KRG.Conclusion:KRG effectively prevents MPP^(+)-induced SH-SY5 Y cell death by regulating cytochrome c release from mitochondria and PINK1/parkin-mediated mitophagy,through regulation of the Bcl-2 family.
基金This work was supported by grants from the National Natural Science Foundation of China(No.81773712,No.81473198)the Foundation for Innovative Research Groups of NSFC(No.81721005)+2 种基金the National Basic Research Program of China(973 Program,No.2014CB744601)Science Fund for Creative Research Groups of the Natural Science Foundation of Hubei Province(No.2015CFA020)PCSIRT(No.IRT13016).
文摘Mitochondrial superoxide overproduction is believed to be responsible for the neurotoxicity associated with neurodegeneration.Mitochondria-targeted antioxidants,such as MitoQ,have emerged as potentially effective antioxidant therapies.Methionine sulfoxide reductase A(MsrA)is a key mitochondrial-localized endogenous antioxidative enzyme and it can scavenge oxidizing species by catalyzing the methionine(Met)-centered redox cycle(MCRC).In this study,we observed that the natural L-Met acted as a good scavenger for antimycin A-induced mitochondrial superoxide overproduction in PC12 cells.This antioxidation was largely dependent on the Met oxidase activity of MsrA.S-methyl-L-cysteine(SMLC),a natural analogue of Met that is abundantly found in garlic and cabbage,could activate the Met oxidase activity of MsrA to scavenge free radicals.Furthermore,SMLC protected against antimycin A-induced mitochondrial membrane depolarization and alleviated 1-methyl-4-phenylpyridinium(MPP+)-induced neurotoxicity.Thus,our data highlighted the possibility for SMLC supplement in the detoxication of mitochondrial damage by activating the Met oxidase activity of MsrA.
基金supported by the China Postdoctoral Science Foundation,No.1630the Natural Science Foundation of Jiangsu Province in China,No.BK2011402+1 种基金the Jiangsu Province Postdoctoral Research Foundation in China,No.1301174Cthe Jiangsu Province Health Department Foundation in China,No.H201361
文摘In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1（SIAH-1） in PC12 cells. 1-Methyl-4-phenylpyridinium（MPP＋） treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression. Treatment with an SIAH-1 antibody decreased m RNA expression levels of α-synuclein, light chain 3 and SIAH-1, but increased E1 m RNA expression. It also increased cell viability. Combined treatment with MPP＋ and rapamycin reduced SIAH-1 and α-synuclein levels. Treatment with SIAH-1 antibody alone diminished α-synuclein immunoreactivity in PC12 cells, and reduced the colocalization of α-synuclein and light chain 3. These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway. Consequently, SIAH-1 may be a potential new therapeutic target for Parkinson’s disease.
基金funded by Coordination for the Improvement of Higher Education Personnel (CAPES,Brazil-Finance Code 001,to LRB)the S?o Paulo Research Foundation(FAPESP,Brazil,project#2018/07366-4)+1 种基金The National Council for Scientific and Technological Development (CNPq,Brazil,project#303006/2018-8,to LRB)a PhD fellowship from FAPESP under Grant Agreement No 2020/02109-3。
文摘The transient receptor potential melastatin 2 is a calcium-permeable cation channel member of the TRP family. Also known as an oxidative stress-activated channel, the transient receptor potential melastatin 2 gating mechanism is dependent on reactive oxygen species. In pathological conditions, transient receptor potential melastatin 2 is overactivated, leading to a Ca~(2+) influx that alters cell homeostasis and promotes cell death. The role of transient receptor potential melastatin 2 in neurodegenerative diseases, including Alzheimer's disease and ischemia, has already been described and reviewed. However, data on transient receptor potential melastatin 2 involvement in Parkinson's disease pathology has emerged only in recent years and the issue lacks review studies that focus specifically on this topic. The present review aims to elucidate the role of the transient receptor potential melastatin 2 channel in Parkinson's disease by reviewing, summarizing, and discussing the in vitro, in vivo, and human studies published until August 2022. Here we describe fourteen studies that evaluated the transient receptor potential melastatin 2 channel in Parkinson's disease. The Parkinson's disease model used, transient receptor potential melastatin 2 antagonist and genetic approaches, and the main outcomes reported were discussed. The studies described transient receptor potential melastatin 2 activation and enhanced expression in different Parkinson's disease models. They also evidenced protective and restorative effects when using transient receptor potential melastatin 2 antagonists, knockout, or silencing. This review provides a literature overview and suggests where there is a need for more research. As a perspective point, this review shows evidence that supports transient receptor potential melastatin 2 as a pharmacological target for Parkinson's disease in the future.
基金This work is supported by the National Natural Science Foundation of China(81774110 and 81573773).
文摘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.
基金National Natural Science Foundation of China Project(No.82060888)Approved Project of Guangxi Health Department(No.Z2016220)Approved Project of Guangxi Administration of Traditional Chinese Medicine(No.GZZC2020107)。
文摘Objective:To investigate the neuroprotective effects of Fructus Mume total flavone(FMF)against cell apoptosis and mitochondrial injury induced by 1-methyl-4-phenylpyridinium(MPP^(+))in human neuroblastoma(SH-SY5Y)cells and explore its molecular mechanisms.Methods:MPP^(+) induced SH-SY5Y cells injury model were established in vitro cell culture,the cells were divided into 5 groups:normal control group,model group(250μmol·L^(-1) MPP^(+)),FMF low-and middle-and high-dose experimental group(10,50,100μmol·L^(-1) FMF).After 72 h administration,4’,6-diamidino-2-phenylindole(DAPI)staining was used to observe the effects of different concentrations of FMF on the morphologic changes of apoptotic cells,the ratio of cell apoptosis was measured by Annexin-FITC/PI double staining.The mitochondrial membrane electro-bit were detected by flow cytometry(FCM).The expression of Bcl-2,Bax and Caspase-3 were detected by Western blot.Results:The results of DAPI staining showed that the injury SH-SY5Y cells induced by MPP+were densely condensed,the nucleus showed nuclear shrinkage,showing an apoptotic characteristic morphology;after 72h of FMF action,the apoptotic morphology of the cells showed different degrees of improvement,and the apoptotic number of SH-SY5Y cells also decreased.Compared with that in the normal control group,the apoptotic rate and of mitochondrial membrane electrobit of SH-SY5Y cells in the model group increased significantly(P<0.01),the expression of Bax and Caspase-3 proteins increased significantly(P<0.01),Bcl-2 protein and the ratio of Bcl-2/Bax decreased significantly(P<0.01).Compared with that in the model group,the apoptotic rate and mitochondrial membrane electro-bit of SH-SY5Y cells in FMF groups(10,50,100μmol·L^(-1))were significantly lower,while Bax and Caspase-3 proteins were significantly lower(P<0.01),and Bcl-2 protein and the ratio of Bcl-2/Bax were significantly higher,with statistically significant difference in FMF middle-and high-dose experimental groups(P<0.01).The results indicated that FMF can decrease the experession level of Bax and Caspase-3 and increase the ratio of Bcl-2/Bax,inhibit MPP+induced apoptosis.Conclusion:FMF improves the damage of SH-SY5Y cells induced by MPP+,and plays a neuroprotective effect by regulating the expressions of related proteins in mitochondrial apoptosis pathway.
基金supported by the National Natural Science Foundation of China(81202814)the Shanghai Municipal Commission of Health and Family Planning(20124y116)the Young Teachers Training Funding Scheme of Shanghai Colleges and Universities,China(zzszy12026)
文摘Echinacoside （ECH） is protective in a mouse model of Parkinson＇ s disease （PD） induced by 1-methyl-4- phenylpyridinium ion （MPP＋）. To investigate the mechanisms involved, SH-SYSY neuroblastoma ceils were treated with MPP＋ or a combination of MPP＋ and ECH, and the expression of ATF3 （activating transcription factor 3）, CHOP （C/EBP-homologous protein）, SCNA （synuclein alpha）, and GDNF （glial cell line-derived neurotrophic factor） was assessed. The results showed that ECH significantly improved cell survival by inhibiting the generation of MPP＋-induced reactive oxygen species （ROS）. In addition, ECH suppressed the ROS and MPP＋- induced expression of apoptotic genes （ATF3, CHOP, and SCNA）. ECH markedly decreased the MPP＋-induced cas- pase-3 activity in a dose-dependent manner. ATF3- knockdown also decreased the CHOP and cleaved caspase- 3 levels and inhibited the apoptosis induced by MPP＋. Interestingly, ECH partially restored the GDNF expression that was down-regulated by MPP＋. ECH also improved dopaminergic neuron survival during MPP＋ treatment and protected these neurons against the apoptosis induced by MPTP. Taken together, these data suggest that the ROS/ ATF3/CHOP pathway plays a critical role in mechanisms by which ECH protects against MPP＋-induced apoptosis in PD.
基金supported by the Innovation and Technology Commission Funding Administrative System of Hong Kong(Project No.:ITS/122/19FP)the Natural Science Fund of Guangdong(project no.2019A1515011257)
文摘Objective:The objective of this study was to investigate whether 70%aqueous ethanol extract of San-Jia-Fu-Mai decoction extract(SJFMDE)could protect against 1-methyl-4-phenylpyridinium(MPP+)-induced oxidative stress in PC12 cells and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced motor function deficits in mice.Materials and Methods:The cell viability,the levels of intracellular reactive oxygen species(ROS),malondialdehyde(MDA),and glutathione(GSH)in the MPP+-treated PC12 cells were measured.Motor function deficits and dopamine(DA)level in the brain striatum and tyrosine hydroxylase(TH)-positive cells in substantia nigra pars compacta(SNc)of the MPTP-treated mice were determined.Results:The results showed that SJFMDE could reduce cell death and the levels of ROS and MDA while increase the level of GSH in the MPP+-treated PC12 cells.In addition,in vivo studies showed that oral administration of SJFMDE(3,6,and 12 g/kg)significantly improved the motor function deficits induced by MPTP and enhanced the DA level in the striatum and TH-positive neuronal cells in SNc of the MPTP-treated mice.Conclusions:Our results revealed that SJFMDE possessed neuroprotective effects against neurotoxicity induced by MPP+and motor function deficits induced by MPTP via suppressing oxidative stress and increasing the levels of DA and TH,indicating that SJFMDE might be a promising Chinese medicine formula for the treatment of Parkinson’s disease.
