In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson＇s Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived mesen...In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson＇s Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived mesenchymal stem cells against 6-hydroxydopamine-induced neurotoxicity and to verify whether the mechanism of action relates to abnormal a-synuclein accumulation in cells Results showed that co-culture with bone marrow-derived mesenchymal stem cells enhanced PC12 cell viability and dopamine secretion in a cell dose-dependent manner. MitoLight staining was used to confirm that PC12 cells co-cultured with bone marrow-derived mesenchymal stem cells demonstrate reduced levels of cell apoptosis. Immunocytochemistry and western blot analysis found the quantity of α-synuclein accumulation was significantly reduced in PC12 cell and bone marrow-derived mesenchymal stem cell co-cultures. These results indicate that bone marrow-derived mesenchymal stem cells can attenuate 6-hydroxydopamine-induced cytotoxicity by reducing abnormal α-synuclein accumulation in PC12 cells.展开更多
Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on m...Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on metabolic and func- tional specialization of the cell bodies versus the axon terminals, the level and disposition of dopamine, its metabolites and enzymes are different in both regions and are likely to be affected differently in PD. We examined changes in the midbrain dopamine system following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), to test the hypothesis that a predisposing/sensitization stage and a inducing/precipitating stage underlie PD. Pregnant mice were treated with a low dose of MPTP during gestation days 8 - 12 to model the predisposing/sensitization stage, by interrupting the fetal mid- brain dopamine system during its neurogenesis. For the inducing/precipitating stage, the 12-weeks offspring were ad- ministered MPTP. The prenatal-MPTP offspring appear normal, but midbrain dopamine, 3,4-di-hydroxy-phenyl-acetic- acid, 3-methoxytyramine, tyrosine-hydroxylase and L-aromatic-amino-acid-decarboxylase, were reduced by 49.6%, 48%, 54%, 20.9% and 25%. Postnatal-MPTP of 10, 20, 30 mg/kg administered to the prenatal-PBS vs prenatal-MPTP offspring reduced midbrain dopamine by 43.6%, 47.2%, 70.3% vs 85.4%, 89.1%, 95.2%;tyrosine-hydroxylase by 30%, 63%, 81% vs 30.7%, 70.4%, 91.4%;L-aromatic-amino-acid-decarboxylase by 0%, 2%, 40% vs 32%, 40%, 58%. The prenatal-MPTP may render the DA system sensitive by causing sub-threshold reduction of DA, its metabolites and en- zymes, enabling postnatal-MPTP to reduce dopamine above the 70% - 80% PD-inducing threshold. Thus, the study may produce a prenatal predisposing/sensitization and postnatal inducing/precipitation model of PD. It also indicates that some cases of PD may have a fetal basis, in which sub-threshold nigrostriatal impairments occur early in life and PD-symptoms are induced during aging by further insults to the dopaminergic system that would not cause PD symptoms in normal indi-viduals.展开更多
Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function,and also regulate pathological processes in brain diseases and injury.However,the role of astrocytes in the dopamine-...Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function,and also regulate pathological processes in brain diseases and injury.However,the role of astrocytes in the dopamine-depleted striatum remains unclear.A rat model of Parkinson’s disease was therefore established by injecting 10μL 6-hydroxydopamine(2.5μg/μL)into the right medial forebrain bundle.Immunohistochemical staining was used to detect the immunoreactivity of glial fibrillary acidic protein(GFAP),calcium-binding protein B(S100B),and signal transducer and activator of transcription 3(STAT3)in the striatum,and to investigate the co-expression of GFAP with S100B and STAT3.Western blot assay was used to measure the protein expression of GFAP,S100B,and STAT3 in the striatum.Results demonstrated that striatal GFAP-immunoreactive cells had an astrocytic appearance under normal conditions,but that dopamine depletion induced a reactive phenotype with obvious morphological changes.The normal striatum also contained S100B and STAT3 expression.S100B-immunoreactive cells were uniform in the striatum,with round bodies and sparse,thin processes.STAT3-immunoreactive cells presented round cell bodies with sparse processes,or were darkly stained with a large cell body.Dopamine deprivation induced by 6-hydroxydopamine significantly enhanced the immunohistochemical positive reaction of S100B and STAT3.Normal striatal astrocytes expressed both S100B and STAT3.Striatal dopamine deprivation increased the number of GFAP/S100B and GFAP/STAT3 double-labeled cells,and increased the protein levels of GFAP,S100B,and STAT3.The present results suggest that morphological changes in astrocytes and changes in expression levels of astrocyte-related proteins are involved in the pathological process of striatal dopamine depletion.The study was approved by Animal Care and Use Committee of Sun Yat-sen University,China(Zhongshan Medical Ethics 2014 No.23)on September 22,2014.展开更多
To prevent and treat Parkinson＇s disease in its early stages,it is essential to be able to detect the degree of early dopaminergic neuron degeneration.Dopamine transporters（DAT） in the striatum regulate synaptic do...To prevent and treat Parkinson＇s disease in its early stages,it is essential to be able to detect the degree of early dopaminergic neuron degeneration.Dopamine transporters（DAT） in the striatum regulate synaptic dopamine levels,and striatal ^99mTc-TRODAT-1 single-photon emission computed tomography（-SPECT） imaging is a marker for presynaptic neuronal degeneration.However,the association between the degree of dopaminergic degeneration and in vivo ^99mTc-TRODAT-1 SPECT imaging is unknown.Therefore,this study investigated the association between the degree of 6-hydroxydopamine（6-OHDA）-induced dopaminergic degeneration and DAT imaging using^99mTc-TRODAT-1 SPECT in rats.Different degrees of nigrostriatal dopamine depletion were generated by injecting different doses of 6-OHDA（2,4,and 8 μg） into the right medial forebrain bundle.The degree of nigrostriatal dopaminergic neuron degeneration was assessed by rotational behavior and immunohistochemical staining.The results showed that striatal ^99mTc-TRODAT-1 binding was significantly diminished both in the ipsilateral and the contralateral sides in the 4 and 8 μg 6-OHDA groups,and that DAT ^99mTc-TRODAT-1 binding in the ipsilateral striatum showed a high correlation to apomorphine-induced rotations at 8 weeks post-lesion（r = –0.887,P 〈 0.01）.There were significant correlations between DAT ^99mTc-TRODAT-1 binding in the ipsilateral striatum and the amount of tyrosine hydroxylase immunoreactive neurons in the ipsilateral substantia nigra in the 2,4,and 8 μg 6-OHDA groups at 8 weeks post-lesion（r = 0.899,P 〈 0.01）.These findings indicate that striatal DAT imaging using ^99mTc-TRODAT-1 is a useful technique for evaluating the severity of dopaminergic degeneration.展开更多
To study the effect of PD Ⅰ administration on dopamine receptors （DR, , DRz ） mRNAs expression in the lesioned striatum of the PD rat model and confirm if PDⅠ has the effect of dopamine receptor agonist. The PD ra...To study the effect of PD Ⅰ administration on dopamine receptors （DR, , DRz ） mRNAs expression in the lesioned striatum of the PD rat model and confirm if PDⅠ has the effect of dopamine receptor agonist. The PD rats with unilateral 6-hydroxydopamine lesioned were administrated with PD Ⅰ , L-dopa methyl/benserazide, L-dopa methyl/benserazide/ PD Ⅰ , normal saline respectively for 4 weeks and their behavioral changes were observed. Then the rats were sacrificed and RT-PCR technique was used to detect changes of dopamine receptors （DR1, DR2） mRNAs expression in the ipsilateral striatum 1 day after the last treatment. The results showed that treatment with PD Ⅰ plus L-dopa resulted in a stable contralateral rotation behavior; treatment with L-dopa resulted in a progressively increased contralateral rotation behavior. Rotation behavior induced by anhydromorphine decreased with PD Ⅰ or PD Ⅰ plus L-dopa treatment. Treatment With L-dopa or PD Ⅰ plus L-dopa, up-regulation of DR, mRNA and down-regulation of DR2 mRNA were observed in the ipsilateral striatum which were more obvious than that treated with PD Ⅰ or vehicle （P〈0. 05）. It was concluded that long-term treatment with PD Ⅰ could alleviate the behavior of PD rats. PD Ⅰ had no apparent effect on the dopamine receptors （DRI , DRz ） mRNAs expression in the ipsilateral striatum and the PD Ⅰ has no agonist effect on dopamine receptors.展开更多
The purpose of this study is to investigate the possible protective mechanism of Buyin Qianzheng Fang(BYQZF)on endoplasmic reticular and mitochondrial function through intracellular Ca^(2+) to protect dopamine neurons...The purpose of this study is to investigate the possible protective mechanism of Buyin Qianzheng Fang(BYQZF)on endoplasmic reticular and mitochondrial function through intracellular Ca^(2+) to protect dopamine neurons in Parkinson's disease mice model.C57BL/6 mice were randomly divided into normal group,model group,BYQZF group.The behavior of mice was observed by climbing and beam hang tests.展开更多
目的建立脑内灌流6-羟基多巴胺(6-OHDA)所致大鼠纹状体细胞外液羟自由基升高的模型,为老年退行性病变和其它氧化应激所致脑细胞损伤的研究和药物筛选提供可利用的方法。方法微透析脑内灌流6-OHDA造模,采用水杨酸捕获羟自由基,高效液相-...目的建立脑内灌流6-羟基多巴胺(6-OHDA)所致大鼠纹状体细胞外液羟自由基升高的模型,为老年退行性病变和其它氧化应激所致脑细胞损伤的研究和药物筛选提供可利用的方法。方法微透析脑内灌流6-OHDA造模,采用水杨酸捕获羟自由基,高效液相-电化学检测技术,对活体脑内羟自由基所形成的2,3二羟基苯甲酸(2,3-DHBA)和2,5二羟基苯甲酸(2,5-DHBA)进行测定。结果6-OHDA脑内灌流后,模型组大鼠纹状体细胞外液2,3-DHBA和2,5-DH-BA在75 m in分别为对照组的6.6和3.4倍;2,3-DHBA在观察的全程中,一直高于对照组(P<0.01);2,5DHBA大部分时间点也高于对照组(P<0.05或P<0.01);维生素EC组的2,3-DHBA有4个时点低于模型组(P<0.05或P<0.01),2,5-DHBA各时点均低于模型组,但差异无显著性。结论6-OHDA脑内灌流可以造成大鼠纹状体细胞外液羟自由基升高的急性模型。展开更多
基金supported by the Science and Technology Development Foundation of Beijing Science and Technology Commission, No. Z101107052210004
文摘In the present study, PC12 cells induced by 6-hydroxydopamine as a model of Parkinson＇s Disease, were used to investigate the protective effects of bone marrow-derived mesenchymal stem cells bone marrow-derived mesenchymal stem cells against 6-hydroxydopamine-induced neurotoxicity and to verify whether the mechanism of action relates to abnormal a-synuclein accumulation in cells Results showed that co-culture with bone marrow-derived mesenchymal stem cells enhanced PC12 cell viability and dopamine secretion in a cell dose-dependent manner. MitoLight staining was used to confirm that PC12 cells co-cultured with bone marrow-derived mesenchymal stem cells demonstrate reduced levels of cell apoptosis. Immunocytochemistry and western blot analysis found the quantity of α-synuclein accumulation was significantly reduced in PC12 cell and bone marrow-derived mesenchymal stem cell co-cultures. These results indicate that bone marrow-derived mesenchymal stem cells can attenuate 6-hydroxydopamine-induced cytotoxicity by reducing abnormal α-synuclein accumulation in PC12 cells.
文摘Dopamine cell bodies in the substantia nigra of the midbrain and with their terminals projecting to the neostriatum form the nigrostriatum and these dopamine neurons degenerate in Parkinson’s disease (PD). Based on metabolic and func- tional specialization of the cell bodies versus the axon terminals, the level and disposition of dopamine, its metabolites and enzymes are different in both regions and are likely to be affected differently in PD. We examined changes in the midbrain dopamine system following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), to test the hypothesis that a predisposing/sensitization stage and a inducing/precipitating stage underlie PD. Pregnant mice were treated with a low dose of MPTP during gestation days 8 - 12 to model the predisposing/sensitization stage, by interrupting the fetal mid- brain dopamine system during its neurogenesis. For the inducing/precipitating stage, the 12-weeks offspring were ad- ministered MPTP. The prenatal-MPTP offspring appear normal, but midbrain dopamine, 3,4-di-hydroxy-phenyl-acetic- acid, 3-methoxytyramine, tyrosine-hydroxylase and L-aromatic-amino-acid-decarboxylase, were reduced by 49.6%, 48%, 54%, 20.9% and 25%. Postnatal-MPTP of 10, 20, 30 mg/kg administered to the prenatal-PBS vs prenatal-MPTP offspring reduced midbrain dopamine by 43.6%, 47.2%, 70.3% vs 85.4%, 89.1%, 95.2%;tyrosine-hydroxylase by 30%, 63%, 81% vs 30.7%, 70.4%, 91.4%;L-aromatic-amino-acid-decarboxylase by 0%, 2%, 40% vs 32%, 40%, 58%. The prenatal-MPTP may render the DA system sensitive by causing sub-threshold reduction of DA, its metabolites and en- zymes, enabling postnatal-MPTP to reduce dopamine above the 70% - 80% PD-inducing threshold. Thus, the study may produce a prenatal predisposing/sensitization and postnatal inducing/precipitation model of PD. It also indicates that some cases of PD may have a fetal basis, in which sub-threshold nigrostriatal impairments occur early in life and PD-symptoms are induced during aging by further insults to the dopaminergic system that would not cause PD symptoms in normal indi-viduals.
基金supported by the National Natural Science Foundation of China,No.81471288(to WLL)the National High Technology Research and Development Program of China(863 Program),No.2017YFA0104704(to WLL)
文摘Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function,and also regulate pathological processes in brain diseases and injury.However,the role of astrocytes in the dopamine-depleted striatum remains unclear.A rat model of Parkinson’s disease was therefore established by injecting 10μL 6-hydroxydopamine(2.5μg/μL)into the right medial forebrain bundle.Immunohistochemical staining was used to detect the immunoreactivity of glial fibrillary acidic protein(GFAP),calcium-binding protein B(S100B),and signal transducer and activator of transcription 3(STAT3)in the striatum,and to investigate the co-expression of GFAP with S100B and STAT3.Western blot assay was used to measure the protein expression of GFAP,S100B,and STAT3 in the striatum.Results demonstrated that striatal GFAP-immunoreactive cells had an astrocytic appearance under normal conditions,but that dopamine depletion induced a reactive phenotype with obvious morphological changes.The normal striatum also contained S100B and STAT3 expression.S100B-immunoreactive cells were uniform in the striatum,with round bodies and sparse,thin processes.STAT3-immunoreactive cells presented round cell bodies with sparse processes,or were darkly stained with a large cell body.Dopamine deprivation induced by 6-hydroxydopamine significantly enhanced the immunohistochemical positive reaction of S100B and STAT3.Normal striatal astrocytes expressed both S100B and STAT3.Striatal dopamine deprivation increased the number of GFAP/S100B and GFAP/STAT3 double-labeled cells,and increased the protein levels of GFAP,S100B,and STAT3.The present results suggest that morphological changes in astrocytes and changes in expression levels of astrocyte-related proteins are involved in the pathological process of striatal dopamine depletion.The study was approved by Animal Care and Use Committee of Sun Yat-sen University,China(Zhongshan Medical Ethics 2014 No.23)on September 22,2014.
基金supported by the National Natural Science Foundation of China,No.81571250
文摘To prevent and treat Parkinson＇s disease in its early stages,it is essential to be able to detect the degree of early dopaminergic neuron degeneration.Dopamine transporters（DAT） in the striatum regulate synaptic dopamine levels,and striatal ^99mTc-TRODAT-1 single-photon emission computed tomography（-SPECT） imaging is a marker for presynaptic neuronal degeneration.However,the association between the degree of dopaminergic degeneration and in vivo ^99mTc-TRODAT-1 SPECT imaging is unknown.Therefore,this study investigated the association between the degree of 6-hydroxydopamine（6-OHDA）-induced dopaminergic degeneration and DAT imaging using^99mTc-TRODAT-1 SPECT in rats.Different degrees of nigrostriatal dopamine depletion were generated by injecting different doses of 6-OHDA（2,4,and 8 μg） into the right medial forebrain bundle.The degree of nigrostriatal dopaminergic neuron degeneration was assessed by rotational behavior and immunohistochemical staining.The results showed that striatal ^99mTc-TRODAT-1 binding was significantly diminished both in the ipsilateral and the contralateral sides in the 4 and 8 μg 6-OHDA groups,and that DAT ^99mTc-TRODAT-1 binding in the ipsilateral striatum showed a high correlation to apomorphine-induced rotations at 8 weeks post-lesion（r = –0.887,P 〈 0.01）.There were significant correlations between DAT ^99mTc-TRODAT-1 binding in the ipsilateral striatum and the amount of tyrosine hydroxylase immunoreactive neurons in the ipsilateral substantia nigra in the 2,4,and 8 μg 6-OHDA groups at 8 weeks post-lesion（r = 0.899,P 〈 0.01）.These findings indicate that striatal DAT imaging using ^99mTc-TRODAT-1 is a useful technique for evaluating the severity of dopaminergic degeneration.
文摘To study the effect of PD Ⅰ administration on dopamine receptors （DR, , DRz ） mRNAs expression in the lesioned striatum of the PD rat model and confirm if PDⅠ has the effect of dopamine receptor agonist. The PD rats with unilateral 6-hydroxydopamine lesioned were administrated with PD Ⅰ , L-dopa methyl/benserazide, L-dopa methyl/benserazide/ PD Ⅰ , normal saline respectively for 4 weeks and their behavioral changes were observed. Then the rats were sacrificed and RT-PCR technique was used to detect changes of dopamine receptors （DR1, DR2） mRNAs expression in the ipsilateral striatum 1 day after the last treatment. The results showed that treatment with PD Ⅰ plus L-dopa resulted in a stable contralateral rotation behavior; treatment with L-dopa resulted in a progressively increased contralateral rotation behavior. Rotation behavior induced by anhydromorphine decreased with PD Ⅰ or PD Ⅰ plus L-dopa treatment. Treatment With L-dopa or PD Ⅰ plus L-dopa, up-regulation of DR, mRNA and down-regulation of DR2 mRNA were observed in the ipsilateral striatum which were more obvious than that treated with PD Ⅰ or vehicle （P〈0. 05）. It was concluded that long-term treatment with PD Ⅰ could alleviate the behavior of PD rats. PD Ⅰ had no apparent effect on the dopamine receptors （DRI , DRz ） mRNAs expression in the ipsilateral striatum and the PD Ⅰ has no agonist effect on dopamine receptors.
文摘The purpose of this study is to investigate the possible protective mechanism of Buyin Qianzheng Fang(BYQZF)on endoplasmic reticular and mitochondrial function through intracellular Ca^(2+) to protect dopamine neurons in Parkinson's disease mice model.C57BL/6 mice were randomly divided into normal group,model group,BYQZF group.The behavior of mice was observed by climbing and beam hang tests.
文摘目的建立脑内灌流6-羟基多巴胺(6-OHDA)所致大鼠纹状体细胞外液羟自由基升高的模型,为老年退行性病变和其它氧化应激所致脑细胞损伤的研究和药物筛选提供可利用的方法。方法微透析脑内灌流6-OHDA造模,采用水杨酸捕获羟自由基,高效液相-电化学检测技术,对活体脑内羟自由基所形成的2,3二羟基苯甲酸(2,3-DHBA)和2,5二羟基苯甲酸(2,5-DHBA)进行测定。结果6-OHDA脑内灌流后,模型组大鼠纹状体细胞外液2,3-DHBA和2,5-DH-BA在75 m in分别为对照组的6.6和3.4倍;2,3-DHBA在观察的全程中,一直高于对照组(P<0.01);2,5DHBA大部分时间点也高于对照组(P<0.05或P<0.01);维生素EC组的2,3-DHBA有4个时点低于模型组(P<0.05或P<0.01),2,5-DHBA各时点均低于模型组,但差异无显著性。结论6-OHDA脑内灌流可以造成大鼠纹状体细胞外液羟自由基升高的急性模型。
