Objective To probe into the function mechanism of penetration therapy with head electrical acupuncture on Parkinson＇s disease. Methods Microinjection of 6-hydroxydopamin （6-OHDA） on the left cor- pus striatum was a...Objective To probe into the function mechanism of penetration therapy with head electrical acupuncture on Parkinson＇s disease. Methods Microinjection of 6-hydroxydopamin （6-OHDA） on the left cor- pus striatum was adopted to prepare rotation model of Parkinson^s disease in rat. Penetration therapy with head electrical acupuncture was administered in treatment. Normal group, sham-operation group, model group and penetration therapy group were set up. （1）lmmunohistochemical （IHC） method was used to test the morphology and count of positive cell of tyrosine hydroxylase （TH）. （2）RT-PCR technology was used to detect the expression of nestin mRNA of neural stem cell （NSC）. Results （1）Compared with model group, in pene- tration therapy group, the expressions of TH-positive neurons in immune response were increased in areal density （AD）, numerical density （ND） and integrating optic density （P〈0.05）. （2）Compared with model group, in penetration therapy group, the expression of nestin mRNA was increased （P〈0. 05）. Conclusion Penetration therapy with head electrical acupuncture promotes the proliferation of endogenous neural stem cells in substantia nigra of rat model of Parkinson＇s disease.展开更多
Objective To evaluate the role of thrombin-activated microglia in the neurodegeneration of nigral dopaminergic neurons in the rat substantia nigra （SN） in vivo. Methods After stereotaxic thrombin injection into unil...Objective To evaluate the role of thrombin-activated microglia in the neurodegeneration of nigral dopaminergic neurons in the rat substantia nigra （SN） in vivo. Methods After stereotaxic thrombin injection into unilateral SN of rats, immunostaining, reverse transcription polymerase chain reaction （RT-PCR） and biochemical methods were used to observe tyrosine hydroxylase （TH） irnmunoreactive positive cells, microglia activation, nitric oxide （NO） amount and inducible nitricoxide synthase （iNOS） expression. Results （1） Selective damage to dopaminergic neurons was produced after thrombin injection, which was evidenced by loss of TH imrnunostaining in time-dependent manner; （2） Strong microglial activation was observed in the SN; （3） RT-PCR demonstrated the early and transient expression of neurotoxic factors iNOS mRNA in the SN. Immunofluorescence results found that thrombin induced expression of iNOS in microglia. The NO production in the thrombininjected rats was significantly higher than that of controls （P 〈 0.05）. Conclusion Thrombin intranigral injection can injure the dopaminergic neurons in the SN. Thrombin-induced microglia activation precedes dopaminergic neuron degeneration, which suggest that activation of microglia and release of NO may play important roles in dopaminergic neuronal death in the SN.展开更多
Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods ...Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods After cultured in vitro for 14 d, the rat organotypic midbrain slices were pretreated with different concentrations （0, 1, 3, 6 or 10 ng/mL） of LPS for 24 h followed by treatment with 100 ng/mL LPS for 72 h. The whole slice viability was detelmined by measurement of the activity of lactic acid dehydrogenase （LDH）. Tyrosine hydroxylase-immunoreactive （TH-IR） neurons and CD 1 1 b/c equivalent-immunoreactive （OX-42-IR） microglia in the slices were observed by immunohistochemical method, and tumor necrosis factor-α （TNF-α levels in the culture media were detected by enzymelinked immunosorbent assays （ELISA）. Results In the slices treated with 100 ng/mL LPS for 72 h, the number of TH-IR neurons reduced from 191± 12 in the control slices to 46±4, and the LDH activity elevated obviously （P 〈 0.01）, along with remarkably increased number of OX-42-IR cells and production of TNF-α （P 〈 0.01）. Preconditioning with 3 or 6 ng/mL LPS attenuated neuron loss （the number of TH-IR neurons increased to 126± 12 and 180± 13, respectively） and markedly reduced LDH levels （P 〈 0.05）, accompanied by significant decreases of OX-42-IR microglia activation and TNF-α production （P 〈 0.05）. Conclusion Low-dose LPS preconditioning could protect dopaminergic neurons against inflammatory damage in rat midbrain slice culture, and inhibition of microglial activation and reduction of the proinflammatory factor TNF-α production may contribute to this protective effect. Further understanding the underlying mechanism of LPS preconditioning may open a new window for treatment of Parkinson＇s disease.展开更多
OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS...OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS Ventral mesencephalic precursors from Ell inbred rat embryos and BMSCs from adult rats were cultured both separately and in co-culture. After a 7-day incubation in vitro, three conditioned culture media were obtained, termed VMP or common medium, BMSC medium, and BMSC±VMP medium. Ventral mesen- cephalic precursors cells were cultured in each of these media and the effects on proliferation and VMP differentiation were assessed. The relative yield of TH± cells was calculated and compared by immunocytochemical staining. RESULTS After a 7-day culture and induction of VMPs, the total cell counts were increased by （44.13±4.75）-fold （common）, （60.63±5.25）-fold （BMSC）, and （64.00±7.63）-fold （BMSC±VMP）. The proportions of TH＋ cells were （18.76±5.20）%, （23.49±4.10）%, and （28.08± 5.42）%, respectively, with statistically significant differences among the treatment groups. CONCLUSION BMSCs release factors that promote the proliferation of VMPs and facilitate the committed differentiation of VMPs into dopaminergic neurons.展开更多
Objective To observe mechanisms of electroacupuncture （EA） on treating Parkinson＇ s disease （PD）. Methods Fifty SD rats were randomly divided into 4 groups： a normal control group （Norm）, a sham operation gro...Objective To observe mechanisms of electroacupuncture （EA） on treating Parkinson＇ s disease （PD）. Methods Fifty SD rats were randomly divided into 4 groups： a normal control group （Norm）, a sham operation group （Sham）, a model group （Mod） and an electroacupuncture （EA） group. EA at ＂Fengfu＂ （风府 GV16） and ＂Taichong＂ （太冲 LR 3） was applied once daily for 2 weeks in Group EA, and no treatment was given to the animals in the other three groups. PD model was established by using 6-hydroxydopamine. Numbers of tyrosine hydroxylase （TH）, tumor necrosis factor-alpha （TNF-α）and interleukin-1 beta （IL-1β） positive reactive cells in substantia nigra （SN） were counted by using immunohistochemical methods. Results The total TH positive neurons in Group Mod were obviously less than those in Group Norm and Group Sham （P〈0.01）, and the numbers of TNF-α and IL-1β positive cells in Group Mod increased more significantly than those in Group Norm, and Group Sham （P〈0.01）. The total TH positive neurons in Group EA were obviously more than those in Group Mod （P〈0.01）, and the numbers of TNF-α and IL-1β positive cells in Group EA were obviously less than those in Group Mod （P〈0.01）, indicating that EA could markedly lower the contents of such pre-inflammatory factors like TNF-α and IL-1β in SN of experimental rats with PD. Conclusion EA may provide its protective functions on dopaminergic neurons by way of alleviating inflammatory reactions in SN of rats with PD.展开更多
Objective To study the apoptotic effects of 1 methyl 4 phenyl 1,2,3,6 tetrahydropyridine (MPTP) on the nigral dopaminergic neurons of mice and 1 methyl 4 phenylpyridium ion (MPP +) on pheochromocytoma (P...Objective To study the apoptotic effects of 1 methyl 4 phenyl 1,2,3,6 tetrahydropyridine (MPTP) on the nigral dopaminergic neurons of mice and 1 methyl 4 phenylpyridium ion (MPP +) on pheochromocytoma (PC12) cells, as well as the antagonism of Eldepryl against MPTP's apoptotic effect Methods Three groups of C 57 BL mice were treated with MPTP, Eldepryl plus MPTP and normal saline, respectively, for 7 days before performing TUNEL (terminal deoxyneucleotidyl transferase mediated dUTP x nick end labeling) and FACS (fluorescence activated cell sorting) analyses of neuronal apoptosis in the substantia nigra The same tests were employed in cell culture to examine apoptosis in PC12 cells treated with MPP +, MPTP or PBS Results Intraperitoneal administration of MPTP 30?mg/kg could induce nigral apoptosis, and oral use of Eldepryl prior to MPTP treatment could completely prevent the nigral apoptosis caused by MPTP MPP +, an intermediate metabolite of MPTP, could lead to the apoptosis of PC12 cells, whereas MPTP itself had no such effect on PC12 cells Conclusions The experiment indicated that the neurotoxin, MPTP, might cause the death of nigral neurons through a mechanism of apoptosis and this effect might be mediated by its bioactive intermediate metabolite MPP + Eldepryl could protect the neurotoxicity from MPTP展开更多
Dopaminergic(DA)neuron-like cells obtained through direct reprogramming of primary human fibroblasts offer exciting opportunities for treatment of Parkinson’s disease.A significant obstacle is the low efficiency of c...Dopaminergic(DA)neuron-like cells obtained through direct reprogramming of primary human fibroblasts offer exciting opportunities for treatment of Parkinson’s disease.A significant obstacle is the low efficiency of conversion during the reprogramming process.Here,we demonstrate that the suppression of p53 significantly enhances the efficiency of transcription factor-mediated conversion of human fibroblasts into functional dopaminergic neurons.In particular,blocking p53 activity using a dominant-negative p53(p53-DN)in IMR90 cells increases the conversion efficiency by 5–20 fold.The induced DA neuron-like cells exhibit dopamine neuron-specific gene expression,significant dopamine uptake and production capacities,and enables symptomatic relief in a rat Parkinson’s disease model.Taken together,our findings suggest that p53 is a critical barrier in direct reprogramming of fibroblast into dopaminergic neurons.展开更多
基金the Excellent Discipline Leadership Fund Project of Harbin Science-Technology Administration :2006RFXYS044
文摘Objective To probe into the function mechanism of penetration therapy with head electrical acupuncture on Parkinson＇s disease. Methods Microinjection of 6-hydroxydopamin （6-OHDA） on the left cor- pus striatum was adopted to prepare rotation model of Parkinson^s disease in rat. Penetration therapy with head electrical acupuncture was administered in treatment. Normal group, sham-operation group, model group and penetration therapy group were set up. （1）lmmunohistochemical （IHC） method was used to test the morphology and count of positive cell of tyrosine hydroxylase （TH）. （2）RT-PCR technology was used to detect the expression of nestin mRNA of neural stem cell （NSC）. Results （1）Compared with model group, in pene- tration therapy group, the expressions of TH-positive neurons in immune response were increased in areal density （AD）, numerical density （ND） and integrating optic density （P〈0.05）. （2）Compared with model group, in penetration therapy group, the expression of nestin mRNA was increased （P〈0. 05）. Conclusion Penetration therapy with head electrical acupuncture promotes the proliferation of endogenous neural stem cells in substantia nigra of rat model of Parkinson＇s disease.
文摘Objective To evaluate the role of thrombin-activated microglia in the neurodegeneration of nigral dopaminergic neurons in the rat substantia nigra （SN） in vivo. Methods After stereotaxic thrombin injection into unilateral SN of rats, immunostaining, reverse transcription polymerase chain reaction （RT-PCR） and biochemical methods were used to observe tyrosine hydroxylase （TH） irnmunoreactive positive cells, microglia activation, nitric oxide （NO） amount and inducible nitricoxide synthase （iNOS） expression. Results （1） Selective damage to dopaminergic neurons was produced after thrombin injection, which was evidenced by loss of TH imrnunostaining in time-dependent manner; （2） Strong microglial activation was observed in the SN; （3） RT-PCR demonstrated the early and transient expression of neurotoxic factors iNOS mRNA in the SN. Immunofluorescence results found that thrombin induced expression of iNOS in microglia. The NO production in the thrombininjected rats was significantly higher than that of controls （P 〈 0.05）. Conclusion Thrombin intranigral injection can injure the dopaminergic neurons in the SN. Thrombin-induced microglia activation precedes dopaminergic neuron degeneration, which suggest that activation of microglia and release of NO may play important roles in dopaminergic neuronal death in the SN.
基金the Foundation of Beijing Municipal Commission of Education,China (No.200410025011)
文摘Objective To identify the protective effect of lipopolysaccharide （LPS） preconditioning against LPS-induced inflammatory damage in dopaminergic neurons of midbrain slice culture and the possible mechanisms. Methods After cultured in vitro for 14 d, the rat organotypic midbrain slices were pretreated with different concentrations （0, 1, 3, 6 or 10 ng/mL） of LPS for 24 h followed by treatment with 100 ng/mL LPS for 72 h. The whole slice viability was detelmined by measurement of the activity of lactic acid dehydrogenase （LDH）. Tyrosine hydroxylase-immunoreactive （TH-IR） neurons and CD 1 1 b/c equivalent-immunoreactive （OX-42-IR） microglia in the slices were observed by immunohistochemical method, and tumor necrosis factor-α （TNF-α levels in the culture media were detected by enzymelinked immunosorbent assays （ELISA）. Results In the slices treated with 100 ng/mL LPS for 72 h, the number of TH-IR neurons reduced from 191± 12 in the control slices to 46±4, and the LDH activity elevated obviously （P 〈 0.01）, along with remarkably increased number of OX-42-IR cells and production of TNF-α （P 〈 0.01）. Preconditioning with 3 or 6 ng/mL LPS attenuated neuron loss （the number of TH-IR neurons increased to 126± 12 and 180± 13, respectively） and markedly reduced LDH levels （P 〈 0.05）, accompanied by significant decreases of OX-42-IR microglia activation and TNF-α production （P 〈 0.05）. Conclusion Low-dose LPS preconditioning could protect dopaminergic neurons against inflammatory damage in rat midbrain slice culture, and inhibition of microglial activation and reduction of the proinflammatory factor TNF-α production may contribute to this protective effect. Further understanding the underlying mechanism of LPS preconditioning may open a new window for treatment of Parkinson＇s disease.
基金This work was supported by grants from Natural Science Foundation of Jiangsu Province （No.BK2004043）
文摘OBJECTIVE To explore the potential neurotrophic effect of bone marrow stromal cells （BMSCs） on cell proliferation and committed neuronal differentiation of ventral mesencephalic precursors （VMPs） in vitro. METHODS Ventral mesencephalic precursors from Ell inbred rat embryos and BMSCs from adult rats were cultured both separately and in co-culture. After a 7-day incubation in vitro, three conditioned culture media were obtained, termed VMP or common medium, BMSC medium, and BMSC±VMP medium. Ventral mesen- cephalic precursors cells were cultured in each of these media and the effects on proliferation and VMP differentiation were assessed. The relative yield of TH± cells was calculated and compared by immunocytochemical staining. RESULTS After a 7-day culture and induction of VMPs, the total cell counts were increased by （44.13±4.75）-fold （common）, （60.63±5.25）-fold （BMSC）, and （64.00±7.63）-fold （BMSC±VMP）. The proportions of TH＋ cells were （18.76±5.20）%, （23.49±4.10）%, and （28.08± 5.42）%, respectively, with statistically significant differences among the treatment groups. CONCLUSION BMSCs release factors that promote the proliferation of VMPs and facilitate the committed differentiation of VMPs into dopaminergic neurons.
基金Supported by National Projects of Natural Science Foundation: 30300461, 30973787, 30973809Project of Science and Technology of Education Department of Hubei Province: 2002 B 00004, D 200726001, Z 20091601+1 种基金Project of Science and Technology of Hubei Science and Technology Agency: 2009 CDA 068, 2010 CDA 101Project of Science and Technology of Wuhan Science and Technology Agency: 201060623269
文摘Objective To observe mechanisms of electroacupuncture （EA） on treating Parkinson＇ s disease （PD）. Methods Fifty SD rats were randomly divided into 4 groups： a normal control group （Norm）, a sham operation group （Sham）, a model group （Mod） and an electroacupuncture （EA） group. EA at ＂Fengfu＂ （风府 GV16） and ＂Taichong＂ （太冲 LR 3） was applied once daily for 2 weeks in Group EA, and no treatment was given to the animals in the other three groups. PD model was established by using 6-hydroxydopamine. Numbers of tyrosine hydroxylase （TH）, tumor necrosis factor-alpha （TNF-α）and interleukin-1 beta （IL-1β） positive reactive cells in substantia nigra （SN） were counted by using immunohistochemical methods. Results The total TH positive neurons in Group Mod were obviously less than those in Group Norm and Group Sham （P〈0.01）, and the numbers of TNF-α and IL-1β positive cells in Group Mod increased more significantly than those in Group Norm, and Group Sham （P〈0.01）. The total TH positive neurons in Group EA were obviously more than those in Group Mod （P〈0.01）, and the numbers of TNF-α and IL-1β positive cells in Group EA were obviously less than those in Group Mod （P〈0.01）, indicating that EA could markedly lower the contents of such pre-inflammatory factors like TNF-α and IL-1β in SN of experimental rats with PD. Conclusion EA may provide its protective functions on dopaminergic neurons by way of alleviating inflammatory reactions in SN of rats with PD.
文摘Objective To study the apoptotic effects of 1 methyl 4 phenyl 1,2,3,6 tetrahydropyridine (MPTP) on the nigral dopaminergic neurons of mice and 1 methyl 4 phenylpyridium ion (MPP +) on pheochromocytoma (PC12) cells, as well as the antagonism of Eldepryl against MPTP's apoptotic effect Methods Three groups of C 57 BL mice were treated with MPTP, Eldepryl plus MPTP and normal saline, respectively, for 7 days before performing TUNEL (terminal deoxyneucleotidyl transferase mediated dUTP x nick end labeling) and FACS (fluorescence activated cell sorting) analyses of neuronal apoptosis in the substantia nigra The same tests were employed in cell culture to examine apoptosis in PC12 cells treated with MPP +, MPTP or PBS Results Intraperitoneal administration of MPTP 30?mg/kg could induce nigral apoptosis, and oral use of Eldepryl prior to MPTP treatment could completely prevent the nigral apoptosis caused by MPTP MPP +, an intermediate metabolite of MPTP, could lead to the apoptosis of PC12 cells, whereas MPTP itself had no such effect on PC12 cells Conclusions The experiment indicated that the neurotoxin, MPTP, might cause the death of nigral neurons through a mechanism of apoptosis and this effect might be mediated by its bioactive intermediate metabolite MPP + Eldepryl could protect the neurotoxicity from MPTP
基金supported in part by grants from the US National Institutes of Health(CA131408,CA136748,CA155270,ES024015)to Li Chuan-Yuan
文摘Dopaminergic(DA)neuron-like cells obtained through direct reprogramming of primary human fibroblasts offer exciting opportunities for treatment of Parkinson’s disease.A significant obstacle is the low efficiency of conversion during the reprogramming process.Here,we demonstrate that the suppression of p53 significantly enhances the efficiency of transcription factor-mediated conversion of human fibroblasts into functional dopaminergic neurons.In particular,blocking p53 activity using a dominant-negative p53(p53-DN)in IMR90 cells increases the conversion efficiency by 5–20 fold.The induced DA neuron-like cells exhibit dopamine neuron-specific gene expression,significant dopamine uptake and production capacities,and enables symptomatic relief in a rat Parkinson’s disease model.Taken together,our findings suggest that p53 is a critical barrier in direct reprogramming of fibroblast into dopaminergic neurons.
