Objective To explore the correlations of transcranial sonography of substantia nigra(SN-TCS)characteristics with MRI iron deposition on substantia nigra in patients with Parkinson disease(PD).Methods Data of SN-TCS an...Objective To explore the correlations of transcranial sonography of substantia nigra(SN-TCS)characteristics with MRI iron deposition on substantia nigra in patients with Parkinson disease(PD).Methods Data of SN-TCS and craniocerebral MRI in 120 PD patients were retrospectively analyzed.The patients were divided into iron deposition positive group(positive group,n=46)and iron deposition negative group(negative group,n=74)according to quantitative susceptibility mapping(QSM)value.Then parameters of SN-TCS and MRI were compared between groups(both P<0.05),and correlation analysis were also performed.Results The proportion of high echo positive,strong echo area and QSM value of substantia nigra,as well as of hyper-substantia nigra area/midbrain area(S/M)in positive group were all higher than those in negative group(all P<0.001).No significant difference of midbrain area was found between groups(P>0.05).Strong echo area of substantia nigra and S/M based on SN-TCS were both low-medium positively correlated with substantia nigra QSM value showed on MRI(r=0.497,0.529,both P<0.001).Conclusion SN-TCS characteristics of PD patients were correlated with MRI iron deposition on substantia nigra,among which strong echo area and S/M were valuable for evaluating iron deposition on substantia nigra.展开更多
The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridin...The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models. Rhizoma Polygonaticould downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustn Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustn Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.展开更多
Levodopa is the gold-standard treatment for Parkinson＇s disease. However, although it alleviates the clinical symptoms, it cannot delay the progressive apoptosis of dopaminergic neurons or prevent motor complications...Levodopa is the gold-standard treatment for Parkinson＇s disease. However, although it alleviates the clinical symptoms, it cannot delay the progressive apoptosis of dopaminergic neurons or prevent motor complications in the long term. In the present study, we investigated the effect of Shudipingchan granule on neuronal apoptosis in a rat model of Parkinson＇s disease, established by injecting 6-hydroxydopamine into the substantia nigra pars compacta and ventral tegmental area. We then administered levodopa （20 mg/kg intraperitoneally, twice daily） with or without Shudipingchan granule （7.5 mL/kg intragastrically, twice daily）, for 4 weeks. The long-term use of levodopa accel- erated apoptosis of nigral cells and worsened behavioral symptoms by activating the extracellular signal-regulated kinase pathway and downstream apoptotic factors. However, administration of Shudipingchan granule with levodopa reduced expression of phosphorylated extracellular signal-regulated kinase 1/2 and Bax, increased tyrosine hydroxylase and Bcl-2, reduced apoptosis in the substantia nigra, and markedly improved dyskinesia. These findings suggest that Shudipingchan granule suppresses neuronal apoptosis by inhibiting the hyper- phosphorylation of extracellular signal-regulated kinase and downregulating expression of anti-apoptotic genes. Shudipingchan granule, used in combination with levodopa, can effectively reduce the symptoms of Parkinson＇s disease.展开更多
Recent researches have found that 7 Tesla SWI can detect the alteration of substantia nigra hyperintensity in Parkinson's disease(PD),multiple system atrophy(MSA),and progressive supranuclear palsy(PSP).The aim of...Recent researches have found that 7 Tesla SWI can detect the alteration of substantia nigra hyperintensity in Parkinson's disease(PD),multiple system atrophy(MSA),and progressive supranuclear palsy(PSP).The aim of this study was to investigate whether 3 Tesla SWI(3T SWI)can visualize anatomical alterations occurring in a hyperintense structure of the substantia nigra in PD and vascular parkinsonism(VP),and whether the evaluation of abnormal signal can be used as a factor in the differential diagnosis of PD and VP.Using 3 Tesla MRI,we evaluated 38 healthy subjects,33 patients with PD and 34 patients with VP.Two blinded readers independently assessed the images.We found that the dorsolateral nigral hyperintensity was absent in 31 of 33 patients with PD and 15 of 34 patients with VP.The dorsolateral nigral hyperintensity was present in 19 of 34 patients with VP and 35 of 38 healthy controls.Group comparisons of absence of dorsolateral nigral hyperintensity revealed significant differences between the patients with PD and those with VP(P<0.001).The sensitivity of SWI for PD was 93.9%and the specificity was 92.1%.Visual assessment of dorsolateral nigral hyperintensity on high-field SWI scans may serve as a new simple diagnostic imaging marker for PD.And our study results indicate that 3T SWI can be used as a tool to identify PD and VP.展开更多
Acupuncture for the treatment of Parkinson＇s disease has a precise clinical outcome. This study investigated the effect of electroacupuncture at Fengfu （GV16） and Taichong （LR3） acupoints in rat models of Parkin...Acupuncture for the treatment of Parkinson＇s disease has a precise clinical outcome. This study investigated the effect of electroacupuncture at Fengfu （GV16） and Taichong （LR3） acupoints in rat models of Parkinson＇s disease induced by subcutaneous injection of rotenone into rat neck and back. Reverse transcription-PCR demonstrated that brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression was significantly increased in the substantia nigra of rat models of Parkinson＇s disease, and that abnormal behavior of rats was significantly improved following electroacupuncture treatment. These results indicated that electroacupuncture treatment upregulated brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression in the substantia nigra of rat models of Parkinson＇s disease. Thus, electroacupuncture may be useful in the treatment of Parkinson＇s disease.展开更多
An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43...An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43（GAP-43） is closely associated with neurite outgrowth and axon regeneration during neural development. We speculate that an enriched environment can reduce damage to dopaminergic neurons by affecting the expression of GAP-43. This study is designed to test this hypothesis. Three-month-old female senescence-accelerated mouse prone 8（SAMP8） mice were housed for 3 months in an enriched environment or a standard environment. These mice were then subcutaneously injected in the abdomen with 14 mg/kg MPTP four times at 2-hour intervals. Morris water maze testing demonstrated that learning and memory abilities were better in the enriched environment group than in the standard environment group. Reverse-transcription polymerase chain reaction, immunohistochemistry and western blot assays showed that m RNA and protein levels of GAP-43 in the substantia nigra were higher after MPTP application in the enriched environment group compared with the standard environment group. These findings indicate that an enriched environment can increase GAP-43 expression in SAMP8 mice. The upregulation of GAP-43 may be a mechanism by which an enriched environment protects against MPTP-induced neuronal damage.展开更多
To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral si...To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67 % vs 6.67 %, P <0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67 %, P >0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P <0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD.展开更多
Because neurons are susceptible to oxidative damage and thioredoxin reductase 1 is extensively distributed in the central nervous system and has antioxidant properties, we speculated that the enzyme may be involved in...Because neurons are susceptible to oxidative damage and thioredoxin reductase 1 is extensively distributed in the central nervous system and has antioxidant properties, we speculated that the enzyme may be involved in the pathogenesis of Parkinson＇s disease. A Parkinson＇s disease model was produced by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine into C57BL/6 mice. Real-time reverse transcription-PCR, western blot analysis and colorimetric assay showed that the levels of thioredoxin reductase 1 mRNA and protein were decreased, along with a significant reduction in thioredoxin reductase activity, in the midbrain of Parkinson＇s disease mice compared with normal mice. Immunohistochemical staining revealed that the number of thioredoxin reductase 1-positive neurons in the substantia nigra pars compacta of Parkinson＇s disease mice was significantly decreased compared with normal mice. These experimental findings suggest that the expression of thioredoxin reductase 1 in the substantia nigra pars compacta of Parkinson＇s disease mice is significantly decreased, and that the enzyme may be associated with disease onset.展开更多
BACKGROUND： To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson＇s disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine （MPTP）, does ...BACKGROUND： To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson＇s disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine （MPTP）, does not exist. In addition, there are no reports of analysis of differential protein expression. OBJECTIVE： To separate and evaluate MPTP-induced differential protein expression through the use of proteomics in the substantia nigra of a mouse model of chronic Parkinson＇s disease. DESIGN： Randomized controlled animal study. SETTING： Department of Neurology, the First Affiliated Hospital, Chongqing Medical University. MATERIALS： Sixteen 8-10-week old, healthy, male, C57BL mice, weighing 20-25 g, and of clean grade, were provided by the Experimental Animal Center of Chongqing Medical University. The experimental animals were disposed according to ethical criteria. MPTP was provided by Sigma Company, USA; Pdquest 2D image analysis software and gelatum/irradiance image analysis system （ChemiDoc XRS） by Bio-Rad, USA; and Voyager DE-PROMALD1-TOF-MS mass spectroscopy analyzer by AB1 Company, USA. METHODS： This study was performed in Chongqing Neurological Laboratory between November 2006 and July 2007. Mice were randomly divided into model and control groups, with 8 mice in each group. Mice in the model group were received a subcutaneous injection of MPTP （25 mg＆g）, twice a week, for five successive weeks, to establish a chronic Parkinson＇s disease model. Mice in the control group received the same volume of a subcutaneous saline injection at the same time points. Mice were sacrificed by anesthesia to rapidly obtain the midbrain for protein separation of the substantia nigra. MAIN OUTCOME MEASURES： （1） 2-ED handbook （Bio-Rad Company） was referenced for two-dimensional electrophoresis, （2） PDQUEST8,0 analytical electrophoresis pattern was adopted to evaluate differential protein expression. （3） Peptide mass finger print map and data were retrieved on http：//www.prospector.ucsf.edu to compare differential substantia nigral protein expression in the two groups. RESULTS： Two-dimensional gel electrophoresis of substantia nigra tissue indicated that there were 33 differential protein expressions between the two groups. Three new proteins were evaluated, including α -enolase, which exhibited regulated expression, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B. CONCLUSION： There are three proteins that exhibit differential expression in the substantia nigra- α -enolase, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B.展开更多
Excessive theta(θ)frequency oscillation and synchronization in the basal ganglia(BG)has been reported in elderly parkinsonian patients and animal models of levodopa(L-dopa)-induced dyskinesia(LID),particularly theθo...Excessive theta(θ)frequency oscillation and synchronization in the basal ganglia(BG)has been reported in elderly parkinsonian patients and animal models of levodopa(L-dopa)-induced dyskinesia(LID),particularly theθoscillation recorded during periods when L-dopa is withdrawn(the off L-dopa state).To gain insight into processes underlying this activity,we explored the relationship between primary motor cortex(M1)oscillatory activity and BG output in LID.We recorded local field potentials in the substantia nigra pars reticulata(SNr)and M1 of awake,inattentive resting rats before and after L-dopa priming in Sham control,Parkinson disease model,and LID model groups.We found that chronic L-dopa increasedθsynchronization and information flow between the SNr and M1 in off L-dopa state LID rats,with a SNr-to-M1 flow directionality.Compared with the on state,θoscillational activity(θsynchronization and informationflow)during the off state were more closely associated with abnormal involuntary movements.Our findings indicate thatθoscillation in M1 may be consequent to abnormal synchronous discharges in the BG and support the notion that M1θoscillation may participate in the induction of dyskinesia.展开更多
BACKGROUND： Following ischemia, apoptosis is observed at the ipsilateral ventropostenor thalamic nucleus and substantia nigra, which are distant from, but connected to, the ischemic cerebral cortex, in animals with n...BACKGROUND： Following ischemia, apoptosis is observed at the ipsilateral ventropostenor thalamic nucleus and substantia nigra, which are distant from, but connected to, the ischemic cerebral cortex, in animals with normotension. However, secondary brain damage in hypertension has not been clearly investigated. OBJECTIVE： The present study determined whether neuronal apoptosis is associated with neuronal loss in the ipsilateral ventroposterior thalamic nucleus and substantia nigra following cortical ischemia in adult hypertensive rats. Results should provide options for determining a time window for anti-apoptotic therapy. DESIGN, TIME AND SETTING： All experimental procedures in this randomized, controlled trial were conducted at the Neurological Laboratory of the First Affiliated Hospital of Sun Yat-sen University of China between October 2006 and July 2008. MATERIALS： Monoclonal primary antibodies specific to mouse anti-rat microtubule-associated protein 2 and glial fibrillary acidic protein were respectively purchased from Sigma Chemical, USA and BD Pharmingen, USA. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling （TUNEL） detection kits were purchased from Roche Applied Science, Switzerland and Upstate, USA, respectively. METHODS： A total of 64 male, Sprague Dawiey rats, aged 60-90 days, were equally and randomly divided into middle cerebral artery occlusion and sham surgery groups. Renovascular hypertension was established in both groups by renal artery occlusion. Right distal middle cerebral artery occlusion was performed by electrocoagulation in the middle cerebral artery occlusion group. MAIN OUTCOME MEASURES： Microtubule-associated protein 2 and glial fibrillary acidic protein were detected by immunohistochemistry, and apoptotic cells were observed by TUNEL assay. The stainings were separately detected in the ipsilateral ventroposterior thalamic nucleus and substantia nigra. RESULTS： During the 4 weeks following distal middle cerebral artery occlusion in renovascular hypertensive rats, microtubule-associated protein 2 expression gradually, but significantly, decreased （P 〈 0.05）. Expression of glial fibrillary acidic protein increased significantly in the ipsilateral ventroposterior thalamic nucleus and substantia nigra （P 〈 0.05） and reached a peak at 4 weeks. In addition, number of apoptotic cells was significantly increased in both areas compared with the sham controls （P 〈 0.05）, with a peak at 2 weeks. CONCLUSION： Results suggested that neuronal loss in the ipsilateral ventroposterior thalamic nucleus and substantia nigra following distal middle cerebral artery occlusion in hypertensive rats could be a secondary event resulting from apoptosis. The temporal apoptosis profile provides options for determining a time window for anti-apoptotic therapy at 2 weeks after stroke.展开更多
Previous studies have shown that Zhichan powder elevated immunity and suppressed oxidation in mice. Rat models of Parkinson＇s disease were induced by stereotaxically injecting 6-hydroxydopamine into the substantia ni...Previous studies have shown that Zhichan powder elevated immunity and suppressed oxidation in mice. Rat models of Parkinson＇s disease were induced by stereotaxically injecting 6-hydroxydopamine into the substantia nigra. The rat models were intragastrically treated with Zhichan powder, which is composed of milkvetch root, ginseng, bunge swallowwort root, himalayan teasel root. Magnolia officinalis, Ligustrum lucidum Ait. and szechwan Iovage rhizome. Immunohistochemistry and reverse transcription-PCR results demonstrated that mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cvtochrome C, Bax, casDase-3, and p53 significantly increased, but Bcl-2 expression significantly decreased in the substantia nigra of rats with Parkinson＇s disease. Following Zhichan powder administration, mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cytochrome C, Bax, caspase-3, and p53 diminished, but Bcl-2 expression increased in the rat substantia nigra. These results indicate that Zhichan powder regulates signal transduction protein expression, inhibits apoptosis, and exerts therapeutic effects on Parkinson＇s disease.展开更多
Histochemistry and EM were used in this experiment to study the effect of EAon the spinal dorsal horn of rat. After 30 minutes EA, the pain threshold of EA rats were elevated400%, the acid phosphatase reaction in spin...Histochemistry and EM were used in this experiment to study the effect of EAon the spinal dorsal horn of rat. After 30 minutes EA, the pain threshold of EA rats were elevated400%, the acid phosphatase reaction in spinal dorsal horn of EA rats increased obviously, the roundagranular vesicles of the C-terminals of EA group was more than that of control’s. The results sug-gested that the histochemical components and numbers of synaptic vesicles in SG and their functionscould be regulated by acupuncture.展开更多
BACKGROUND： Previous researches suggested that expression level of calbindin D28K mRNA decreased in substantia nigra （SN） of model rats with Parkinson disease （PD）, and this might be related to the decrease of an...BACKGROUND： Previous researches suggested that expression level of calbindin D28K mRNA decreased in substantia nigra （SN） of model rats with Parkinson disease （PD）, and this might be related to the decrease of anti-degeneration potentials of dopaminergic neurons. OBJECTIVE： To observe expression changes of calbindin D28K in SN dopaminergic neurons during their degeneration and death in midbrain of PD model rats. DESIGN： Completely randomized grouping design SETTING： Department of Neurobiology, Xuzhou Medical College MATERIALS＂ A total of 92 healthy male SD rats, with the age of 3 months, weighing 200-250 g, were selected from Experimental Animal Center of Xuzhou Medical College [certification： SCXK （su） 2003-0003]. Calbindin D28K（CB）, tyroxine hydroxylase （TH）, ABC kit, 6-hydroxydopamine （6-OHDA） and Nissl dyes were provided by Sigma Company, and sheep serum was provided by Beijing Zhongshan Company. METHODS： The experiment was carried out in the Neurobiological Center of Xuzhou Medical College from October 2003 to October 2004. ① With lot method, rats were divided into blank control group （n=28）, experimental control group （n=-28） and experimental group （n=36）. Rats in experimental group were injected with 6-OHDA at right corpus striatum for PD modeling; rats in experimental control group were injected with saline at the same site; rats in blank control group did not give any injections.② On the 7^th, 14^th, 21^st and 28^th days, SN segments on right midbrain from every 5 rats in experimental group were fixed, embedded with paraffin and cut into successively coronary pieces. Rats in other two groups were treated with the same methods and then stained with Nissl to show neuronal form. Meanwhile, CB and TH antibodies staining with immunohistochemistry were used to show CB containing dopaminergic neurons and dopaminergic neurons, and cells were calculated and observed under optic microscope. ③ On the 14^th and 28^th days, every 4 rats in experimental group and every 4 rats in control group were selected to obtain their brains and separate SN on the injured side. Western blot was used to detect expression of calbindin D28K, protein band was scanned with imaging equipment, and data were analyzed with LabWorks software. MAIN OUTCOME MEASURES：① On the 7^th, 14^th, 21^st and 28^th days, Nissl staining results of SN neurons and immunohistochemical staining results of CB and TH antibodies; ② On the 14^th and 28^th days, Western blot results of calbindin D28K in SN neurons, RESULTS; Among 92 rats, 2 rats in experimental group died after 1 day due to 6-OHDA injection and other 90 rats were involved in the final analysis. ①Nissl staining results： On the 7^th day of 6-OHDA injection, most neuronal somas on right SN pars compacta were shown as deep pycnosis or lysis breakage; on the 14^th and 21^st days, amount of neurons was decreased remarkably; on the 28^th day, most neurons in SN pars compacta disappeared. ② Results of immunohistochemical staining： Amount of positive neurons of calbindin D28K in right SN pars compacta was not changed on the 7^th day after 6-OHDA injection; on the 14^th day, the amount was higher in experimental group than that in both control groups （P 〈 0.01） and was decreased till the 21^st day, but it was still higher than that in the two control groups （P 〈 0.05）; on the 28^th day, positive neurons of calbindin D28K nearly disappeared, and the amount was lower than that in the two control groups （P 〈 0.01）. In addition, on the 7^th day after 6-OHDA injection into corpus striatum, positive TH neurons decreased 24% in right SN, and there was significant difference from that in control groups; on the 14^th, 21^st and 28^th days, positive TH neurons decreased 37%, 46% and 64%, respectively. ③ Western blot results： On the 14＇h day after 6-OHDA injection into corpus striatum, expression of calbindin D28K in right SN was higher in experimental group than that in both control groups （P 〈 0.05）; however, on the 28^th day, the expression was lower than that in the two control groups （P 〈 0.01 ）. CONCLUSION ： During degeneration and death of dopaminergic neurons, CB expression in SN pars compacta increases firstly and decreased gradually.展开更多
To date, complex components of available reagents have been used for directional induction of neural stem cells into dopaminergic neurons, resulting in a poor ability to repeat experiments. This study sought to invest...To date, complex components of available reagents have been used for directional induction of neural stem cells into dopaminergic neurons, resulting in a poor ability to repeat experiments. This study sought to investigate whether a homogenate of the substantia nigra of adult rats and/or basic fibroblast growth factor could directionally induce neural stem cells derived from the subventricular zone of embryonic rats to differentiate into dopaminergic neurons. Tyrosine hydroxylase-positive cells were observed exclusively after induction with the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor for 48 hours in vitro. However, in the groups treated with homogenate supernatant or basic fibroblast growth factor alone, tyrosine hydroxylase expression was not observed. Moreover, the content of dopamine in the culture medium of subventricular zone neurons was significantly increased at 48 hours after induction with the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor. Experimental findings indicate that the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor could directionally induce neural stem cells derived from the subventricular zone of embryonic rats to differentiate into dopaminergic neurons in the substantia nigra with the ability to secrete dopamine.展开更多
Parkinson＇s disease is a progressive neurological disease characterized by the degeneration of dopaminergic neurons in the substantia nigra.A highly similar pattern of neurodegeneration can be induced by 1-methyl-4-p...Parkinson＇s disease is a progressive neurological disease characterized by the degeneration of dopaminergic neurons in the substantia nigra.A highly similar pattern of neurodegeneration can be induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP） or 1-methyl-4-phenylpyridinium ion（MPP＋）,which cause the death of dopaminergic neurons.Administration of MPTP or MPP＋ results in Parkinson＇s disease-like symptoms in rodents.However,it remains unclear whether intracerebroventricular MPP＋ administration affects neurogenesis in the substantia nigra and subgranular zone or whether brain-derived neurotrophic factor alters the effects of MPP＋.In this study,MPP＋（100 nmol） was intracerebroventricularly injected into mice to model Parkinson＇s disease.At 7 days after administration,the number of bromodeoxyuridine（Brd U）-positive cells in the subgranular zone of the hippocampal dentate gyrus increased,indicating enhanced neurogenesis.In contrast,a reduction in Brd U-positive cells was detected in the substantia nigra.Administration of brain-derived neurotrophic factor（100 ng） 1 day after MPP＋ administration attenuated the effect of MPP＋ in the subgranular zone and the substantia nigra.These findings reveal the complex interaction between neurotrophic factors and neurotoxins in the Parkinsonian model that result in distinct effects on the catecholaminergic system and on neurogenesis in different brain regions.展开更多
BACKGROUND： Cellular adhesion molecule mediates the effects of glial cell line-derived neurotrophic factor in dopaminergic neurons during nervous system development. It is assumed that cellular adhesion molecule play...BACKGROUND： Cellular adhesion molecule mediates the effects of glial cell line-derived neurotrophic factor in dopaminergic neurons during nervous system development. It is assumed that cellular adhesion molecule plays an important role in dopaminergic neuron development. OBJECTIVE： To investigate N-cadherin expression in the midbrain substantia nigra during rat development. DESIGN, TIME AND SETTING： The developmental biology, controlled study was performed at the Laboratory of Research Center of Neurobiology, Xuzhou Medical College, China from April to October 2007. MATERIALS： Sprague Dawley rats （embryonic days 16, 18, and 20, as well as postnatal days 1, 3, 7, 14, and 21） were selected for this study. N-cadherin antibody was purchased from Santa Cruz, USA. METHODS： N-cadherin expression in the midbrain substantia nigra of perinatal rats was detected using reverse transcription polymerase chain reaction, Western blot, and immunohistochemistry methods. N-cadherin-positive cells were counted. MAIN OUTCOME MEASURES： N-cadherin mRNA, protein, and positive cells in the rat midbrain substantia nigra were quantified. RESULTS： The results of reverse transcription polymerase chain reaction and Western blot demonstrated that altered N-cadherin mRNA levels were similar to protein levels in the midbrain substantia nigra. N-cadherin mRNA and protein expression was low in the midbrain substantia nigra of embryonic day 16 rats but gradually increased, and reached a peak at postnatal day 1. N-cadherin mRNA and protein levels were still high at postnatal days 3 and 7, but significantly decreased at postnatal day 14. There was no significant difference in N-cadherin mRNA and protein expression between postnatal days 14 and 21 （P 〉 0.05）. The immunohistochemistry results demonstrated that there was no significant difference in the quantity of N-cadherin-positive cells per area in rats from embryonic day 16 to postnatal day 7 （P 〉 0.05）. The quantity of N-cadherin-positive cells per area was significantly decreased in rats at postnatal day 14. There was no significant difference in the number of N-cadherin-positive cells per area between postnatal days 14 and 21 （P 〉 0.05）. CONCLUSION： N-cadherin expression was reduced during embryonic stages, but reached a peak at postnatal days 1-7, and then gradually decreased in the rat midbrain substantia nigra.展开更多
The corneal sutotantia propria in guinea pig is composed of the fibroblasts,fibers andintercelular maxtrix.Tab fibers belong to collagen fibrils.The fibrils are fine(about 37nm in diameter)and form lamelae being paral...The corneal sutotantia propria in guinea pig is composed of the fibroblasts,fibers andintercelular maxtrix.Tab fibers belong to collagen fibrils.The fibrils are fine(about 37nm in diameter)and form lamelae being parallel to one another.The lamelae are approximately 0.75￣1.5μmthick. The fibrils in adjacent lamelae run across each other at right angles and parallel to the cornealsurface.in fibroblasts are dispersed between the fibril lamelae. Their appearances are extremelyflat. The nucleus is large and located at the canter of the cell.The cytoplasm is abundant in roughendoplasmic reticulum.The corned fibroblasts give out very long processes extended between thelamelae.The cytoplasmic brocades of a cell are in able contant with these of neighboring cells.Thecontant surfaces show irregular and interdigital forms. In some of the intercellular contant regionsthere are gap junctions.展开更多
The substantia nigra pars reticulate(SNr),which plays a pivotal role in motor control,is the key structure in integrating information for cortex,basal ganglia and thalamus.Abnormal gait and posture deficits can be rev...The substantia nigra pars reticulate(SNr),which plays a pivotal role in motor control,is the key structure in integrating information for cortex,basal ganglia and thalamus.Abnormal gait and posture deficits can be reversed by SNr deep brain stimulation(DBS)in certain Parkinson’s disease cases.However,functional characterization of SNr,which is the key for the optimization of DBS effect,remains elusive.In current study,we recorded extracellular single unit in SNr of urethane anesthetized rats.We have found that urethane can induce slow delta and theta oscillations in SNr local field potential.The high gamma oscillation observed is positively correlated with the occurrence of action potential.The putative GABAergic neurons have a mean firing rate of(20.82±2.04)Hz,of which 65.2%display a regular firing pattern and 34.8%show irregular firing.Our results demonstrated the heterogeneous property of SNr and provided possible theoretical basis for promoting the next generation of DBS electrode design and optimization of clinical DBS parameters.展开更多
Background:A century ago,a mystery between a virus and Parkinson's disease(PD)was described.Owing to the limitation of human brain biopsy and the challenge of electron microscopy in observing virions in human brai...Background:A century ago,a mystery between a virus and Parkinson's disease(PD)was described.Owing to the limitation of human brain biopsy and the challenge of electron microscopy in observing virions in human brain tissue,it has been difficult to study the viral etiology of PD.Recent discovery of virobiota reveals that viruses coexist with humans as symbionts.Newly developed transcriptomic sequencing and novel bioinformatic approaches for mining the encrypted virome in human transcriptome make it possible to study the relationship between symbiotic viruses and PD.Nevertheless,whether viruses exist in the human substantia nigra(SN)and whether symbiotic viruses underlie PD pathogenesis remain unknown.Methods:We collected current worldwide human SN transcriptomic datasets from the United States,the United Kingdom,the Netherlands,and Switzerland.We used bioinformatic approaches including viruSITE and the Viral-Track to identify the existence of viruses in the SN of patients.The comprehensive RNA sequencing-based virome analysis pipeline was used to characterize the virobiota in the SN.The Pearson's correlation analysis was used to examine the association between the viral RNA fragment counts(VRFCs)and PD-related human gene sequencing reads in the SN.The differentially expressed genes(DEGs)in the SN between PD patients and non-PD individuals were used to examine the molecular signatures of PD and also evaluate the impact of symbiotic viruses on the SN.Findings:We observed the existence of viruses in the human SN.A dysbiosis of virobiota was found in the SN of PD patients.A marked correlation between VRFC and PD-related human gene expression was detected in the SN of PD patients.These PD-related human genes correlated to VRFC were named as the virus-correlated PD-related genes(VPGs).We identified 3 bacteriophages(phages),including the Proteus phage VB_PmiS-Isfahan,the Escherichia phage phix174,and the Lactobacillus phage Shal,that might impair the gene expression of neural cells in the SN of PD patients.The Proteus phage VB_PmiS-lsfahan was a common virus in the SN of patients from the United Kingdom,the Netherlands,and Switzerland.VPGs and DEGs together highlighted that the phages might dampen dopamine biosynthesis and weaken the cGAS-STING function.Interpretation:This is the first study to discover the involvement of phages in PD pathogenesis.A lifelong low symbiotic viral load in the SN may be a contributor to PD pathogenesis.Our findings unlocked the black box between brain virobiota and PD,providing a novel insight into PD etiology from the perspective of phage-human symbiosis.展开更多
文摘Objective To explore the correlations of transcranial sonography of substantia nigra(SN-TCS)characteristics with MRI iron deposition on substantia nigra in patients with Parkinson disease(PD).Methods Data of SN-TCS and craniocerebral MRI in 120 PD patients were retrospectively analyzed.The patients were divided into iron deposition positive group(positive group,n=46)and iron deposition negative group(negative group,n=74)according to quantitative susceptibility mapping(QSM)value.Then parameters of SN-TCS and MRI were compared between groups(both P<0.05),and correlation analysis were also performed.Results The proportion of high echo positive,strong echo area and QSM value of substantia nigra,as well as of hyper-substantia nigra area/midbrain area(S/M)in positive group were all higher than those in negative group(all P<0.001).No significant difference of midbrain area was found between groups(P>0.05).Strong echo area of substantia nigra and S/M based on SN-TCS were both low-medium positively correlated with substantia nigra QSM value showed on MRI(r=0.497,0.529,both P<0.001).Conclusion SN-TCS characteristics of PD patients were correlated with MRI iron deposition on substantia nigra,among which strong echo area and S/M were valuable for evaluating iron deposition on substantia nigra.
基金supported by the Natural Science Foundation of Fujian Province, No. 2009J06018
文摘The Chinese herbs Herba Epimedii, Fructus Ligustri Lucidi and Rhizoma Polygonati were injected into Parkinson＇s disease mice established via intraperitoneal injection of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride. The selective monoamine oxidase B inhibitor selegiline was used as a positive control drug. After successive administration for 4 weeks, Herba Epimedii could downregulate the expression of caspase-3 and increase the brain-derived neurotrophic factor level, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models. Rhizoma Polygonaticould downregulate the expression of caspase-3 and FasL, and increase neural growth factor and brain-derived neurotrophic factor levels. Fructus Ligustn Lucidi could downregulate caspase-3 expression. Rhizoma Polygonati and Fructus Ligustn Lucidi did not produce obvious effects on tyrosine hydroxylase activity. Herba Epimedii and Fructus Ligustri Lucidi yielded similar effects on apoptosis-promoting factors to those elicited by selegiline. Herba Epimedii and Rhizoma Polygonati significantly increased the levels of neurotrophic factors compared with selegiline. Herba Epimedii significantly increased tyrosine hydroxylase activity compared with selegiline. It is indicated that the kidney-tonifying Chinese herbal preparation can downregulate the expression of apoptosis-promoting factors, increase neurotrophic factors levels in the substantia nigra and striatum, as well as increase tyrosine hydroxylase activity in the substantia nigra of Parkinson＇s disease mouse models, thereby exerting a stronger or similar neuroprotective effects compared with selegiline.
基金financially supported by the National Natural Science Foundation of China,No.81302926,30472207the Major Project of Shanghai Science and Technology Commission of China,No.15401970100the Shanghai Municipal Commission of Health and Family Planning of China,No.ZY3-RCPY-2-2005
文摘Levodopa is the gold-standard treatment for Parkinson＇s disease. However, although it alleviates the clinical symptoms, it cannot delay the progressive apoptosis of dopaminergic neurons or prevent motor complications in the long term. In the present study, we investigated the effect of Shudipingchan granule on neuronal apoptosis in a rat model of Parkinson＇s disease, established by injecting 6-hydroxydopamine into the substantia nigra pars compacta and ventral tegmental area. We then administered levodopa （20 mg/kg intraperitoneally, twice daily） with or without Shudipingchan granule （7.5 mL/kg intragastrically, twice daily）, for 4 weeks. The long-term use of levodopa accel- erated apoptosis of nigral cells and worsened behavioral symptoms by activating the extracellular signal-regulated kinase pathway and downstream apoptotic factors. However, administration of Shudipingchan granule with levodopa reduced expression of phosphorylated extracellular signal-regulated kinase 1/2 and Bax, increased tyrosine hydroxylase and Bcl-2, reduced apoptosis in the substantia nigra, and markedly improved dyskinesia. These findings suggest that Shudipingchan granule suppresses neuronal apoptosis by inhibiting the hyper- phosphorylation of extracellular signal-regulated kinase and downregulating expression of anti-apoptotic genes. Shudipingchan granule, used in combination with levodopa, can effectively reduce the symptoms of Parkinson＇s disease.
文摘Recent researches have found that 7 Tesla SWI can detect the alteration of substantia nigra hyperintensity in Parkinson's disease(PD),multiple system atrophy(MSA),and progressive supranuclear palsy(PSP).The aim of this study was to investigate whether 3 Tesla SWI(3T SWI)can visualize anatomical alterations occurring in a hyperintense structure of the substantia nigra in PD and vascular parkinsonism(VP),and whether the evaluation of abnormal signal can be used as a factor in the differential diagnosis of PD and VP.Using 3 Tesla MRI,we evaluated 38 healthy subjects,33 patients with PD and 34 patients with VP.Two blinded readers independently assessed the images.We found that the dorsolateral nigral hyperintensity was absent in 31 of 33 patients with PD and 15 of 34 patients with VP.The dorsolateral nigral hyperintensity was present in 19 of 34 patients with VP and 35 of 38 healthy controls.Group comparisons of absence of dorsolateral nigral hyperintensity revealed significant differences between the patients with PD and those with VP(P<0.001).The sensitivity of SWI for PD was 93.9%and the specificity was 92.1%.Visual assessment of dorsolateral nigral hyperintensity on high-field SWI scans may serve as a new simple diagnostic imaging marker for PD.And our study results indicate that 3T SWI can be used as a tool to identify PD and VP.
基金supported by the National Natural Science Foundation of China,No.30973787,30973809the Open Research Fund of Zhejiang First-foremost Key Subject-Acupuncture & Moxibustion,No.ZTK2010A10+1 种基金the Natural Science Foundation of Hubei Province,No.2009CDA068the Integrated Traditional and Western Medicine project by the Health Department of Hubei Province,No.2010Z-Z01
文摘Acupuncture for the treatment of Parkinson＇s disease has a precise clinical outcome. This study investigated the effect of electroacupuncture at Fengfu （GV16） and Taichong （LR3） acupoints in rat models of Parkinson＇s disease induced by subcutaneous injection of rotenone into rat neck and back. Reverse transcription-PCR demonstrated that brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression was significantly increased in the substantia nigra of rat models of Parkinson＇s disease, and that abnormal behavior of rats was significantly improved following electroacupuncture treatment. These results indicated that electroacupuncture treatment upregulated brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression in the substantia nigra of rat models of Parkinson＇s disease. Thus, electroacupuncture may be useful in the treatment of Parkinson＇s disease.
基金supported by a grant from the Health Department of Hebei Province of China,No.20120056,20140314the Funding Project for Introduced Abroad Study Personnel of Hebei Province of China,No.C2011003039
文摘An enriched environment protects dopaminergic neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP）-induced neuronal injury, but the underlying mechanism for this is not clear. Growth associated protein-43（GAP-43） is closely associated with neurite outgrowth and axon regeneration during neural development. We speculate that an enriched environment can reduce damage to dopaminergic neurons by affecting the expression of GAP-43. This study is designed to test this hypothesis. Three-month-old female senescence-accelerated mouse prone 8（SAMP8） mice were housed for 3 months in an enriched environment or a standard environment. These mice were then subcutaneously injected in the abdomen with 14 mg/kg MPTP four times at 2-hour intervals. Morris water maze testing demonstrated that learning and memory abilities were better in the enriched environment group than in the standard environment group. Reverse-transcription polymerase chain reaction, immunohistochemistry and western blot assays showed that m RNA and protein levels of GAP-43 in the substantia nigra were higher after MPTP application in the enriched environment group compared with the standard environment group. These findings indicate that an enriched environment can increase GAP-43 expression in SAMP8 mice. The upregulation of GAP-43 may be a mechanism by which an enriched environment protects against MPTP-induced neuronal damage.
文摘To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67 % vs 6.67 %, P <0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67 %, P >0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P <0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD.
基金supported by the Fundamental Research Funds for the Central Universities, No. lzujbky-2011-83Project of the International Cooperation and Communion Department of Chinese Education Ministry (46th batch)the Science Foundation of Key Laboratory of Preclinical Study for New Drugs of Gansu Province, No. GSKFKT-0804
文摘Because neurons are susceptible to oxidative damage and thioredoxin reductase 1 is extensively distributed in the central nervous system and has antioxidant properties, we speculated that the enzyme may be involved in the pathogenesis of Parkinson＇s disease. A Parkinson＇s disease model was produced by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine into C57BL/6 mice. Real-time reverse transcription-PCR, western blot analysis and colorimetric assay showed that the levels of thioredoxin reductase 1 mRNA and protein were decreased, along with a significant reduction in thioredoxin reductase activity, in the midbrain of Parkinson＇s disease mice compared with normal mice. Immunohistochemical staining revealed that the number of thioredoxin reductase 1-positive neurons in the substantia nigra pars compacta of Parkinson＇s disease mice was significantly decreased compared with normal mice. These experimental findings suggest that the expression of thioredoxin reductase 1 in the substantia nigra pars compacta of Parkinson＇s disease mice is significantly decreased, and that the enzyme may be associated with disease onset.
基金the National Natural Science Foundation of China, No. 30370499
文摘BACKGROUND： To date, a complete protein expression profile of the midbrain substantia nigra in a mouse model of chronic Parkinson＇s disease, induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine （MPTP）, does not exist. In addition, there are no reports of analysis of differential protein expression. OBJECTIVE： To separate and evaluate MPTP-induced differential protein expression through the use of proteomics in the substantia nigra of a mouse model of chronic Parkinson＇s disease. DESIGN： Randomized controlled animal study. SETTING： Department of Neurology, the First Affiliated Hospital, Chongqing Medical University. MATERIALS： Sixteen 8-10-week old, healthy, male, C57BL mice, weighing 20-25 g, and of clean grade, were provided by the Experimental Animal Center of Chongqing Medical University. The experimental animals were disposed according to ethical criteria. MPTP was provided by Sigma Company, USA; Pdquest 2D image analysis software and gelatum/irradiance image analysis system （ChemiDoc XRS） by Bio-Rad, USA; and Voyager DE-PROMALD1-TOF-MS mass spectroscopy analyzer by AB1 Company, USA. METHODS： This study was performed in Chongqing Neurological Laboratory between November 2006 and July 2007. Mice were randomly divided into model and control groups, with 8 mice in each group. Mice in the model group were received a subcutaneous injection of MPTP （25 mg＆g）, twice a week, for five successive weeks, to establish a chronic Parkinson＇s disease model. Mice in the control group received the same volume of a subcutaneous saline injection at the same time points. Mice were sacrificed by anesthesia to rapidly obtain the midbrain for protein separation of the substantia nigra. MAIN OUTCOME MEASURES： （1） 2-ED handbook （Bio-Rad Company） was referenced for two-dimensional electrophoresis, （2） PDQUEST8,0 analytical electrophoresis pattern was adopted to evaluate differential protein expression. （3） Peptide mass finger print map and data were retrieved on http：//www.prospector.ucsf.edu to compare differential substantia nigral protein expression in the two groups. RESULTS： Two-dimensional gel electrophoresis of substantia nigra tissue indicated that there were 33 differential protein expressions between the two groups. Three new proteins were evaluated, including α -enolase, which exhibited regulated expression, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B. CONCLUSION： There are three proteins that exhibit differential expression in the substantia nigra- α -enolase, tumor necrosis factor ligand superfamily member 4, and cyclin-dependent kinase inhibitor 1B.
基金supported by the National Natural Science Foundation of China(81771210)the Natural Science Foundation of Guangdong Province,China(2015A030313288)。
文摘Excessive theta(θ)frequency oscillation and synchronization in the basal ganglia(BG)has been reported in elderly parkinsonian patients and animal models of levodopa(L-dopa)-induced dyskinesia(LID),particularly theθoscillation recorded during periods when L-dopa is withdrawn(the off L-dopa state).To gain insight into processes underlying this activity,we explored the relationship between primary motor cortex(M1)oscillatory activity and BG output in LID.We recorded local field potentials in the substantia nigra pars reticulata(SNr)and M1 of awake,inattentive resting rats before and after L-dopa priming in Sham control,Parkinson disease model,and LID model groups.We found that chronic L-dopa increasedθsynchronization and information flow between the SNr and M1 in off L-dopa state LID rats,with a SNr-to-M1 flow directionality.Compared with the on state,θoscillational activity(θsynchronization and informationflow)during the off state were more closely associated with abnormal involuntary movements.Our findings indicate thatθoscillation in M1 may be consequent to abnormal synchronous discharges in the BG and support the notion that M1θoscillation may participate in the induction of dyskinesia.
基金Chinese Medical Board of USA, No.CMB00-730the Na-tional Natural Science Foundation of China, No. 30770764, 30973108the Fund of Health Department of Guangdong Province Department of China, No.A2009172
文摘BACKGROUND： Following ischemia, apoptosis is observed at the ipsilateral ventropostenor thalamic nucleus and substantia nigra, which are distant from, but connected to, the ischemic cerebral cortex, in animals with normotension. However, secondary brain damage in hypertension has not been clearly investigated. OBJECTIVE： The present study determined whether neuronal apoptosis is associated with neuronal loss in the ipsilateral ventroposterior thalamic nucleus and substantia nigra following cortical ischemia in adult hypertensive rats. Results should provide options for determining a time window for anti-apoptotic therapy. DESIGN, TIME AND SETTING： All experimental procedures in this randomized, controlled trial were conducted at the Neurological Laboratory of the First Affiliated Hospital of Sun Yat-sen University of China between October 2006 and July 2008. MATERIALS： Monoclonal primary antibodies specific to mouse anti-rat microtubule-associated protein 2 and glial fibrillary acidic protein were respectively purchased from Sigma Chemical, USA and BD Pharmingen, USA. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling （TUNEL） detection kits were purchased from Roche Applied Science, Switzerland and Upstate, USA, respectively. METHODS： A total of 64 male, Sprague Dawiey rats, aged 60-90 days, were equally and randomly divided into middle cerebral artery occlusion and sham surgery groups. Renovascular hypertension was established in both groups by renal artery occlusion. Right distal middle cerebral artery occlusion was performed by electrocoagulation in the middle cerebral artery occlusion group. MAIN OUTCOME MEASURES： Microtubule-associated protein 2 and glial fibrillary acidic protein were detected by immunohistochemistry, and apoptotic cells were observed by TUNEL assay. The stainings were separately detected in the ipsilateral ventroposterior thalamic nucleus and substantia nigra. RESULTS： During the 4 weeks following distal middle cerebral artery occlusion in renovascular hypertensive rats, microtubule-associated protein 2 expression gradually, but significantly, decreased （P 〈 0.05）. Expression of glial fibrillary acidic protein increased significantly in the ipsilateral ventroposterior thalamic nucleus and substantia nigra （P 〈 0.05） and reached a peak at 4 weeks. In addition, number of apoptotic cells was significantly increased in both areas compared with the sham controls （P 〈 0.05）, with a peak at 2 weeks. CONCLUSION： Results suggested that neuronal loss in the ipsilateral ventroposterior thalamic nucleus and substantia nigra following distal middle cerebral artery occlusion in hypertensive rats could be a secondary event resulting from apoptosis. The temporal apoptosis profile provides options for determining a time window for anti-apoptotic therapy at 2 weeks after stroke.
文摘Previous studies have shown that Zhichan powder elevated immunity and suppressed oxidation in mice. Rat models of Parkinson＇s disease were induced by stereotaxically injecting 6-hydroxydopamine into the substantia nigra. The rat models were intragastrically treated with Zhichan powder, which is composed of milkvetch root, ginseng, bunge swallowwort root, himalayan teasel root. Magnolia officinalis, Ligustrum lucidum Ait. and szechwan Iovage rhizome. Immunohistochemistry and reverse transcription-PCR results demonstrated that mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cvtochrome C, Bax, casDase-3, and p53 significantly increased, but Bcl-2 expression significantly decreased in the substantia nigra of rats with Parkinson＇s disease. Following Zhichan powder administration, mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cytochrome C, Bax, caspase-3, and p53 diminished, but Bcl-2 expression increased in the rat substantia nigra. These results indicate that Zhichan powder regulates signal transduction protein expression, inhibits apoptosis, and exerts therapeutic effects on Parkinson＇s disease.
文摘Histochemistry and EM were used in this experiment to study the effect of EAon the spinal dorsal horn of rat. After 30 minutes EA, the pain threshold of EA rats were elevated400%, the acid phosphatase reaction in spinal dorsal horn of EA rats increased obviously, the roundagranular vesicles of the C-terminals of EA group was more than that of control’s. The results sug-gested that the histochemical components and numbers of synaptic vesicles in SG and their functionscould be regulated by acupuncture.
基金Scientific and Technological Project of State Ministry of Education, No. 204055 the Natural Science Foundation of Educational Bureau of Jiangsu Province, No. 02KJB310009
文摘BACKGROUND： Previous researches suggested that expression level of calbindin D28K mRNA decreased in substantia nigra （SN） of model rats with Parkinson disease （PD）, and this might be related to the decrease of anti-degeneration potentials of dopaminergic neurons. OBJECTIVE： To observe expression changes of calbindin D28K in SN dopaminergic neurons during their degeneration and death in midbrain of PD model rats. DESIGN： Completely randomized grouping design SETTING： Department of Neurobiology, Xuzhou Medical College MATERIALS＂ A total of 92 healthy male SD rats, with the age of 3 months, weighing 200-250 g, were selected from Experimental Animal Center of Xuzhou Medical College [certification： SCXK （su） 2003-0003]. Calbindin D28K（CB）, tyroxine hydroxylase （TH）, ABC kit, 6-hydroxydopamine （6-OHDA） and Nissl dyes were provided by Sigma Company, and sheep serum was provided by Beijing Zhongshan Company. METHODS： The experiment was carried out in the Neurobiological Center of Xuzhou Medical College from October 2003 to October 2004. ① With lot method, rats were divided into blank control group （n=28）, experimental control group （n=-28） and experimental group （n=36）. Rats in experimental group were injected with 6-OHDA at right corpus striatum for PD modeling; rats in experimental control group were injected with saline at the same site; rats in blank control group did not give any injections.② On the 7^th, 14^th, 21^st and 28^th days, SN segments on right midbrain from every 5 rats in experimental group were fixed, embedded with paraffin and cut into successively coronary pieces. Rats in other two groups were treated with the same methods and then stained with Nissl to show neuronal form. Meanwhile, CB and TH antibodies staining with immunohistochemistry were used to show CB containing dopaminergic neurons and dopaminergic neurons, and cells were calculated and observed under optic microscope. ③ On the 14^th and 28^th days, every 4 rats in experimental group and every 4 rats in control group were selected to obtain their brains and separate SN on the injured side. Western blot was used to detect expression of calbindin D28K, protein band was scanned with imaging equipment, and data were analyzed with LabWorks software. MAIN OUTCOME MEASURES：① On the 7^th, 14^th, 21^st and 28^th days, Nissl staining results of SN neurons and immunohistochemical staining results of CB and TH antibodies; ② On the 14^th and 28^th days, Western blot results of calbindin D28K in SN neurons, RESULTS; Among 92 rats, 2 rats in experimental group died after 1 day due to 6-OHDA injection and other 90 rats were involved in the final analysis. ①Nissl staining results： On the 7^th day of 6-OHDA injection, most neuronal somas on right SN pars compacta were shown as deep pycnosis or lysis breakage; on the 14^th and 21^st days, amount of neurons was decreased remarkably; on the 28^th day, most neurons in SN pars compacta disappeared. ② Results of immunohistochemical staining： Amount of positive neurons of calbindin D28K in right SN pars compacta was not changed on the 7^th day after 6-OHDA injection; on the 14^th day, the amount was higher in experimental group than that in both control groups （P 〈 0.01） and was decreased till the 21^st day, but it was still higher than that in the two control groups （P 〈 0.05）; on the 28^th day, positive neurons of calbindin D28K nearly disappeared, and the amount was lower than that in the two control groups （P 〈 0.01）. In addition, on the 7^th day after 6-OHDA injection into corpus striatum, positive TH neurons decreased 24% in right SN, and there was significant difference from that in control groups; on the 14^th, 21^st and 28^th days, positive TH neurons decreased 37%, 46% and 64%, respectively. ③ Western blot results： On the 14＇h day after 6-OHDA injection into corpus striatum, expression of calbindin D28K in right SN was higher in experimental group than that in both control groups （P 〈 0.05）; however, on the 28^th day, the expression was lower than that in the two control groups （P 〈 0.01 ）. CONCLUSION ： During degeneration and death of dopaminergic neurons, CB expression in SN pars compacta increases firstly and decreased gradually.
基金sponsored by the General Program of Yunnan Provincial Application Basic Research, No. 2008ZC109M
文摘To date, complex components of available reagents have been used for directional induction of neural stem cells into dopaminergic neurons, resulting in a poor ability to repeat experiments. This study sought to investigate whether a homogenate of the substantia nigra of adult rats and/or basic fibroblast growth factor could directionally induce neural stem cells derived from the subventricular zone of embryonic rats to differentiate into dopaminergic neurons. Tyrosine hydroxylase-positive cells were observed exclusively after induction with the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor for 48 hours in vitro. However, in the groups treated with homogenate supernatant or basic fibroblast growth factor alone, tyrosine hydroxylase expression was not observed. Moreover, the content of dopamine in the culture medium of subventricular zone neurons was significantly increased at 48 hours after induction with the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor. Experimental findings indicate that the homogenate supernatant of the substantia nigra from adult rats and basic fibroblast growth factor could directionally induce neural stem cells derived from the subventricular zone of embryonic rats to differentiate into dopaminergic neurons in the substantia nigra with the ability to secrete dopamine.
基金supported by the National Natural Science Foundation of China,No.31320103906(to TB)
文摘Parkinson＇s disease is a progressive neurological disease characterized by the degeneration of dopaminergic neurons in the substantia nigra.A highly similar pattern of neurodegeneration can be induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine（MPTP） or 1-methyl-4-phenylpyridinium ion（MPP＋）,which cause the death of dopaminergic neurons.Administration of MPTP or MPP＋ results in Parkinson＇s disease-like symptoms in rodents.However,it remains unclear whether intracerebroventricular MPP＋ administration affects neurogenesis in the substantia nigra and subgranular zone or whether brain-derived neurotrophic factor alters the effects of MPP＋.In this study,MPP＋（100 nmol） was intracerebroventricularly injected into mice to model Parkinson＇s disease.At 7 days after administration,the number of bromodeoxyuridine（Brd U）-positive cells in the subgranular zone of the hippocampal dentate gyrus increased,indicating enhanced neurogenesis.In contrast,a reduction in Brd U-positive cells was detected in the substantia nigra.Administration of brain-derived neurotrophic factor（100 ng） 1 day after MPP＋ administration attenuated the effect of MPP＋ in the subgranular zone and the substantia nigra.These findings reveal the complex interaction between neurotrophic factors and neurotoxins in the Parkinsonian model that result in distinct effects on the catecholaminergic system and on neurogenesis in different brain regions.
基金the National Natural Science Foundation of China,No.3570564
文摘BACKGROUND： Cellular adhesion molecule mediates the effects of glial cell line-derived neurotrophic factor in dopaminergic neurons during nervous system development. It is assumed that cellular adhesion molecule plays an important role in dopaminergic neuron development. OBJECTIVE： To investigate N-cadherin expression in the midbrain substantia nigra during rat development. DESIGN, TIME AND SETTING： The developmental biology, controlled study was performed at the Laboratory of Research Center of Neurobiology, Xuzhou Medical College, China from April to October 2007. MATERIALS： Sprague Dawley rats （embryonic days 16, 18, and 20, as well as postnatal days 1, 3, 7, 14, and 21） were selected for this study. N-cadherin antibody was purchased from Santa Cruz, USA. METHODS： N-cadherin expression in the midbrain substantia nigra of perinatal rats was detected using reverse transcription polymerase chain reaction, Western blot, and immunohistochemistry methods. N-cadherin-positive cells were counted. MAIN OUTCOME MEASURES： N-cadherin mRNA, protein, and positive cells in the rat midbrain substantia nigra were quantified. RESULTS： The results of reverse transcription polymerase chain reaction and Western blot demonstrated that altered N-cadherin mRNA levels were similar to protein levels in the midbrain substantia nigra. N-cadherin mRNA and protein expression was low in the midbrain substantia nigra of embryonic day 16 rats but gradually increased, and reached a peak at postnatal day 1. N-cadherin mRNA and protein levels were still high at postnatal days 3 and 7, but significantly decreased at postnatal day 14. There was no significant difference in N-cadherin mRNA and protein expression between postnatal days 14 and 21 （P 〉 0.05）. The immunohistochemistry results demonstrated that there was no significant difference in the quantity of N-cadherin-positive cells per area in rats from embryonic day 16 to postnatal day 7 （P 〉 0.05）. The quantity of N-cadherin-positive cells per area was significantly decreased in rats at postnatal day 14. There was no significant difference in the number of N-cadherin-positive cells per area between postnatal days 14 and 21 （P 〉 0.05）. CONCLUSION： N-cadherin expression was reduced during embryonic stages, but reached a peak at postnatal days 1-7, and then gradually decreased in the rat midbrain substantia nigra.
文摘The corneal sutotantia propria in guinea pig is composed of the fibroblasts,fibers andintercelular maxtrix.Tab fibers belong to collagen fibrils.The fibrils are fine(about 37nm in diameter)and form lamelae being parallel to one another.The lamelae are approximately 0.75￣1.5μmthick. The fibrils in adjacent lamelae run across each other at right angles and parallel to the cornealsurface.in fibroblasts are dispersed between the fibril lamelae. Their appearances are extremelyflat. The nucleus is large and located at the canter of the cell.The cytoplasm is abundant in roughendoplasmic reticulum.The corned fibroblasts give out very long processes extended between thelamelae.The cytoplasmic brocades of a cell are in able contant with these of neighboring cells.Thecontant surfaces show irregular and interdigital forms. In some of the intercellular contant regionsthere are gap junctions.
基金the Science and Technology Commission of Shanghai Municipality(No.18JC1413100)the National Key Research and Development Program(No.2016YFC0906400)。
文摘The substantia nigra pars reticulate(SNr),which plays a pivotal role in motor control,is the key structure in integrating information for cortex,basal ganglia and thalamus.Abnormal gait and posture deficits can be reversed by SNr deep brain stimulation(DBS)in certain Parkinson’s disease cases.However,functional characterization of SNr,which is the key for the optimization of DBS effect,remains elusive.In current study,we recorded extracellular single unit in SNr of urethane anesthetized rats.We have found that urethane can induce slow delta and theta oscillations in SNr local field potential.The high gamma oscillation observed is positively correlated with the occurrence of action potential.The putative GABAergic neurons have a mean firing rate of(20.82±2.04)Hz,of which 65.2%display a regular firing pattern and 34.8%show irregular firing.Our results demonstrated the heterogeneous property of SNr and provided possible theoretical basis for promoting the next generation of DBS electrode design and optimization of clinical DBS parameters.
基金supported by grants from the Noncommunicable Chronic Diseases-National Science and Technology Major Project(2024ZD0530200 and 2024ZD0530201 to R.Z.)the National Natural Science Foundation of China(nos.82170864,81471064,81670779,and 81870590 to R.Z.)+3 种基金the National Key Research and Development Program of China(no.2017YFC1700402 to R.Z.)the Beijing Municipal Natural Science Foundation(nos.7162097 and H2018206641 to R.Z.)the Peking University Research Foundation(no.BMU20140366 to R.Z.)the Scientific Project of Beijing Life Science Academy(no.2023300CB0100 to R.Z.).
文摘Background:A century ago,a mystery between a virus and Parkinson's disease(PD)was described.Owing to the limitation of human brain biopsy and the challenge of electron microscopy in observing virions in human brain tissue,it has been difficult to study the viral etiology of PD.Recent discovery of virobiota reveals that viruses coexist with humans as symbionts.Newly developed transcriptomic sequencing and novel bioinformatic approaches for mining the encrypted virome in human transcriptome make it possible to study the relationship between symbiotic viruses and PD.Nevertheless,whether viruses exist in the human substantia nigra(SN)and whether symbiotic viruses underlie PD pathogenesis remain unknown.Methods:We collected current worldwide human SN transcriptomic datasets from the United States,the United Kingdom,the Netherlands,and Switzerland.We used bioinformatic approaches including viruSITE and the Viral-Track to identify the existence of viruses in the SN of patients.The comprehensive RNA sequencing-based virome analysis pipeline was used to characterize the virobiota in the SN.The Pearson's correlation analysis was used to examine the association between the viral RNA fragment counts(VRFCs)and PD-related human gene sequencing reads in the SN.The differentially expressed genes(DEGs)in the SN between PD patients and non-PD individuals were used to examine the molecular signatures of PD and also evaluate the impact of symbiotic viruses on the SN.Findings:We observed the existence of viruses in the human SN.A dysbiosis of virobiota was found in the SN of PD patients.A marked correlation between VRFC and PD-related human gene expression was detected in the SN of PD patients.These PD-related human genes correlated to VRFC were named as the virus-correlated PD-related genes(VPGs).We identified 3 bacteriophages(phages),including the Proteus phage VB_PmiS-Isfahan,the Escherichia phage phix174,and the Lactobacillus phage Shal,that might impair the gene expression of neural cells in the SN of PD patients.The Proteus phage VB_PmiS-lsfahan was a common virus in the SN of patients from the United Kingdom,the Netherlands,and Switzerland.VPGs and DEGs together highlighted that the phages might dampen dopamine biosynthesis and weaken the cGAS-STING function.Interpretation:This is the first study to discover the involvement of phages in PD pathogenesis.A lifelong low symbiotic viral load in the SN may be a contributor to PD pathogenesis.Our findings unlocked the black box between brain virobiota and PD,providing a novel insight into PD etiology from the perspective of phage-human symbiosis.
