ABSTRACT
ABSTRACT Background: The role of autonomic nervous system in the development and maintenance of portal hypertension is not fully elucidated. It is known that the gene expression of norepinephrine in the superior mesenteric artery varies with time, and it may contribute for splanchnic vasodilation and its consequent hemodynamic repercussions. It is still not known exactly how the adrenergic expression behaves at the heart level in the initial stages of this process. Aim: To evaluate the immunohistochemical expression of the enzyme tyrosine hydroxylase (tyrosine 3-monooxygenase), involved in the synthesis of norepinephrine, in the myocardium of rats submitted to partial ligation of the portal vein. Methods: Twenty-four Wistar rats were divided into two groups: Sham Operated and Portal Hypertension. The partial ligation was performed in the Portal Hypertension group, and after 1/6/24 h and 3/5/14 days the animals were euthanized. Immunohistochemical analysis was performed to quantify the expression of the stained enzyme using the ImageJ program. Results: The Portal Hypertension group expressed percentages between 4.6-6% of the marked area, while the Sham Operated group varied between 4-5%. Although there was no statistical significance, the percentage stained in the Portal Hypertension group followed an increasing pattern in the first 6 h and a decreasing pattern after 24 h, which was not observed in the Sham Operated group. Conclusion: The expression of noradrenaline in rat myocardium during the first two weeks after partial ligation of the portal vein, with tyrosine hydroxylase as marker, did not show differences between groups over time.
RESUMO Racional: O papel do sistema nervoso autônomo na hipertensão portal não está completamente elucidado. Sabe-se que, nessa condição, a expressão gênica da norepinefrina na artéria mesentérica superior modifica-se com o tempo, podendo ser importante contribuinte para a vasodilatação esplâncnica e suas repercussões hemodinâmicas. Apesar dos estudos sobre as repercussões cardiovasculares na hipertensão portal, ainda não se sabe como a expressão adrenérgica se comporta a nível cardíaco nas etapas iniciais desse processo. Objetivo: Avaliar a expressão imunoistoquímica da enzima tirosina hidroxilase (tirosina 3-mono-oxigenase), relacionada à síntese da norepinefrina, no miocárdio de ratos submetidos à ligadura parcial da veia porta. Métodos: Foram utilizados 24 ratos, distribuídos em dois grupos: Sham Operated e Hipertensão Portal. A ligadura parcial da veia porta foi realizada apenas no grupo Hipertensão Portal e, após 1/6/24 h e 3/5/14 dias, os animais foram eutanasiados. Foi feita a análise imunoistoquímica para quantificar a expressão da enzima corada, utilizando o programa ImageJ. Resultados: No grupo Hipertensão Portal, o miocárdio expressou percentuais entre 4,6-6% de área marcada, enquanto que no grupo Sham Operated variou entre 4-5%, sem significância estatística. Apenas no grupo Hipertensão Portal, a porcentagem corada pela enzima seguiu padrão crescente nas primeiras 6 h e decrescente após 24 h. Conclusão: A expressão da noradrenalina no miocárdio de ratos durante as primeiras duas semanas após a ligadura parcial da veia porta, tendo como marcador a enzima tirosina hidroxilase, não apresentou diferenças entre grupos ao longo do tempo.
Subject(s)
Animals , Male , Rats , Norepinephrine/biosynthesis , Hypertension, Portal/etiology , Myocardium/metabolism , Tyrosine 3-Monooxygenase/biosynthesis , Catecholamines/physiology , Norepinephrine/physiology , Rats, Wistar , Disease Models, AnimalABSTRACT
PURPOSE: The purpose of this study is to investigate the clinical significance of tyrosine hydroxylase (TH) expression in peripheral blood (PB) at diagnosis in patients with neuroblastoma. MATERIALS AND METHODS: TH mRNA expression in PB was measured by reverse transcription quantitative real-time polymerase chain reaction in 210 patients who were newly diagnosed with neuroblastoma from July 2005 to June 2015 and the clinical significance of TH expression in PB at diagnosis was evaluated. RESULTS: TH expression was positive in 60 patients (28.6%). Fifty of 60 TH-positive patients had metastatic tumors and the remaining 10 had localized tumors. TH expression was associated with high-risk features (i.e., advanced stage, older age, unfavorable pathology, and MYCN amplification) at diagnosis. Among TH-positive patients, higher TH expression level was observed in high-risk patients than in low- or intermediate-risk patients (p=0.035). The probability of 5-year progression-free survival (PFS) was lower in TH-positive patients than in TH-negative patients (63.8±6.9% vs. 94.7±2.1%, p < 0.001). In analysis confined to high-risk patients, the 5-year probability of PFS remained lower in TH-positive patients (55.7±8.2% vs. 89.6±5.8%, p < 0.001). Among TH-positive patients, a higher expression level of TH was associated with a worse outcome. In multivariate analyses, positive TH expression in PB at diagnosis was an independent poor prognostic factor for PFS. CONCLUSION: The treatment intensity should be tailored according to TH expression in PB at diagnosis.
Subject(s)
Humans , Diagnosis , Disease-Free Survival , Multivariate Analysis , Neuroblastoma , Pathology , Polymerase Chain Reaction , Real-Time Polymerase Chain Reaction , Reverse Transcription , RNA, Messenger , Tyrosine 3-Monooxygenase , TyrosineABSTRACT
Objective To observe the effects of Eldepryl on expressions of tyrosine hydroxylase (TH) and glial cell line-derived neurotrophic factor (GDNF) in substantia nigra and striatum in Parkinson’s disease (PD) and to explore the protective mechanism of Eldepryl on dopaminergic neuron . Methods Healthy male Sprague-Dawley (SD) rats (n=72) were randomly divided into control group, model group and Eldepryl group (n=24 in each group). Each group was divided random?ly into 2 subgroups as 4 day treatment group and 8 day treatment group (n=12 in each subgrop). Pakinson’s disease model was established by injecting rotenone subcutaneously back the neck, rats in the control group were injected with an equal vol?ume of sunflower oil subcutaneously at the same location. Rats in the Eldepryl group were then given Eldepryl 0.5 mg·kg-1 in?tragastrically every day for 4 or 8 consecutive days and rats in model group and control group were given an equal volume of saline instead. The expression of TH and GDNF in substantia nigra and striatum were detected by immunohistochemistry and Western blotting. Results Immunohistochemistry and Western blotting showed that strong expression of TH positive cells with little expression of GDNF positive cells were seen in substantia nigra and striatum in rats of control group, and there was no significant difference between subgroup of 8 day treatment and 4 day treatment within control group. The expression of TH cells and GDNF were both significantly reduced in model group compared with those in control group (both P<0.05), and there was no significant difference between subgroup of 8 day treatment and 4 day treatment within each group. The ex?pression of TH positive cells were significantly reduced in Eldepryl group compared with those in control group, and were sig?nificantly increased compared with those in model group. The expression of GDNF positive cells were significantly increased in Eldepryl group compared with those in control group and model group (all P<0.05). And there were significantly more ex?pression of TH positive cells and GDNF positive cells at subgroup of 8 day treatment compared with those at subgroup of 4 day treatment within Eldepryl group with (all P<0.05). Conclusion These data suggest that Eldepryl can protect the dam?age of dopaminergic neurons in substantia nigra and striatum of PD rats. And its therapeutic mechanism may be associated with increased expression of GDNF.
ABSTRACT
BACKGROUND:Stem cel therapy is superior to drug therapy for recovery of patient’s physiological mode, and cel transplantation therapy is becoming a trend. OBJECTIVE:To observe the changes in dopamine content in the stratum of Parkinson’s disease rats after transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cel s. METHODS:After identification by enzyme digestion, pEGFP-C2-TH plasmid was transfected into the fourth generation of human umbilical cord blood mesenchymal stem cel s by electroporation method, and then transfected cel s were injected into the right cerebral ventricle of Parkinson’s disease rats (experimental group). PBS injection was performed in the control group. Migration of dopamine in the brain tissue of rats was observed, and the content of dopamine was detected by high performance liquid chromatography. RESULTS AND CONCLUSION:At 8 weeks after cel transplantation, the cel s gradual y migrated to the ventricles;after 12 weeks, the cel s migrated to the cortex, and expressed tyrosine hydroxylase antigen. Meanwhile, the content of dopamine was significantly higher in the experimental group than the control group (P<0.05). These results reveal that the intraventricular transplantation of tyrosine hydroxylase-modified human umbilical cord blood mesenchymal stem cel s has obvious therapeutic effect on Parkinson’s disease rats.
ABSTRACT
Objective To investigate the efficiency of levodopa in dopa responsive dystonia (DRD) patients and drugs safety in pregnancy cases confirmed by genetic detection.Methods The clinical characteristics of two patients were analyzed.Direct sequences were performed in guanosine triphosphate (GTP) cyclohydrolase Ⅰ (GCH1) gene and tyrosine hydroxylase (TH) gene mutation screening.Results Case 1 was a young man exhibiting writer's cramp and dystonia of lower legs with marked diurnal fluctuation.Writer's cramp could not be relieved by treatment of low dose levodopa/benserazide.After increasing dose,the symptom of writer's cramp appeared occasionally.Case 2 was a young woman who experienced gait disorder.The symptom disappeared completely by levodopa treatment.She used levodopa and benzhexol during pregnancy.By 38 gestational weeks,she gave birth to a healthy baby.Sequence analysis of GCH1 gene in case 1 revealed a mutation (c.230C ＞ G p.S77C) that is a novel pathogenic mutation.The confirmed mutation c.628delC (p.His210Thrfs* 5) found in case 2 had been reported previously.No mutations in TH gene were detected in two patients.Condnsions Most of DRD patients have dramatic response to levodopa,but patients exhibited writer' s cramp may respond to levodopa incompletely.The previous reports indicate that no adverse events have been reported in DRD pregnant women with the monotherapy of levodopa.
ABSTRACT
Objective To explore the role of Th17-and Treg-derived catecholamines during collagen-induced ar?thritis (CIA) progression. Methods Eighteen male DBA/1 mice were randomly divided into control group, CIA model groupⅠ(day 35) and CIA model groupⅡ(day 55). The CIA models were induced by typeⅡcollagen (CⅡ) injection from tails. mRNA expression of Th17 specific transcription factor include ROR-γt, cytokines, IL-17 IL-22, Treg specific transcription factor, Foxp3, cytokines, TGF-βand tyrosine hydroxylase (TH) in lymph nodes were examined by real-time PCR. Co-local?ization of ROR-γt or Foxp3, with TH, vesicular monoamine transporter-2 (VMAT-2) or monoamine oxidase (MAO) in lymph nodes were observed by immunofluorescence staining. Results In lymph nodes of mice in CIAⅠgroup and CIAⅡgroup, mRNA expression of ROR-γt, IL-17, TH and IL-22 were upregulated, while mRNA expression of Foxp3 and TGF-βex?pression was downregulated compared to those expression in control group. The upregulated expression of IL-17 was signifi?cantly reduced in CIAⅡgroup compared with that in CIAⅠgroup. In the lymph nodes of both intact and CIA mice, co-lo?calization of ROR-γt or Foxp3 with TH, VMAT-2 or MAO was seen in some cells. The numbers of cells that are double-pos?itive of ROR-γt/TH,ROR-γt/VMAT-2 and ROR-γt/MAO IL-17 were increased in CIA groups compared to those in con?trol group. And they are significantly reduced in CIAⅡgroup compared with those in CIAⅠgroup. Conclusion The abili?ty to synthesize catecholamines in Th17 cells was increased in lymph node in mice from CIA groups compared to that in con?trol group. The increased catecholamines production from Th17 cells in lymph nodes may be involved in the anti-inflammato?ry progression in CIA.
ABSTRACT
Objective To investigate the clinical characteristics,treatment effect,long-term follow up results,guanosine triphosphate (GTP) cyrclohydrolase Ⅰ (GCH Ⅰ)gene and tyrosine hydroxylase(TH) gene mutations in patients with dopa-responsive dystonia (DRD).Methods The clinical features of 3 families with 4 affected members were analyzed and all of 4 patients were screened for mutations of the GCH Ⅰ gene and TH gene with DNA sequences.Results Four patients were females,average age at onset was (15.3 ± 5.6) years (range:from 9 to 20 years).The initial symptoms were a gait disorder,stiffness or tremor of the lower limbs in all patients presented with diurnal fluctuation.As the increase of disease duration,bilateral hand tremor was found in three patients,systemic torsion was found in one patient and torticollis was found in one patient.All patients' symptoms were in complete remission after administration of low dose of levodopa.Four patients were followed up for 0.5 to 10.0 years,and all were still responsive to the levodopa treatment and effective dosage was decreased as the increase of the disease duration.No longterm side effects of levodopa had occurred after long-term treatment.One patient was found to have c.607G ＞A(p,Gly203Arg) heterogenetic mutation in GCH I gene.Molecular analysis revealed a compound heterozygous mutation in the TH gene (p.Y447Ter and p.V468M) in one patient.No point mutations in both genes were found in other patients.Conclusions DRD patients have dramatic and sustained response to levodopa and no long-term side effects of levodopa after long-term treatment.The detection of GCH Ⅰ and TH gene mutations is helpful in early diagnosis but the negative results could not exclude the diagnosis of DRD.
ABSTRACT
Objective To construct genetically engineered cells that secrete human TH and GDNF at the same time by stable co-transfection pcDNA3.0/hTH and pcDNA3.1/hGDNF in SH-SY5Y cells and study their effects on the gene therapy of Parkinson′s disease (PD). Methods pcDNA3.0/hTH and pcDNA3.1/hGDNF were constructed by inserting human TH and GDNF cDNA into pcDNA3.0 and pcDNA3.1, respectively. SH-SY5Y cells were transfected with pcDNA3.0/hTH and pcDNA3.1/hGDNF and the positive cell clones of human TH and GDNF cDNA engineered cells could be identified by RT-PCR. The engineered cells were co-cultured with primary dopaminergic neurons in mouse. The number and growth condition of primary dopaminergic neurons were examined by Immunocytochemistry. Results The number of primary dopaminergic neurons protected by double-gene engineered cells increased at least by 2.8 times in comparison with the control cells(P
ABSTRACT
Objective To investigate the effect of repeated psychological stress on central nerve dopamine system, and the effect of tyrosine intervention. Methods Rats were subjected to psychological stress by Communication Box model continuously for 14 days (30min/d), and fed with tyrosine in the dose of 250mg/kg, 500mg/kg or 1000mg/kg respectively. Tyrosine hydroxylase (TH) and fos protein of the brain tissue in the ventral tegumental area (VTA), nucleus accumbens (Nac) and mesoprefrontal cortex (mPFC) area were detected by immunohistochemical staining. Results In the ventral tegumental area (VTA), 5.1mm posterior to anterior fontanel, TH positive neurons were obviously less in number in psychological stress group than that in control group (P