Correlación genotipo-fenotipo cardiovascular de la dopamina B- hidroxilasa (DBH) / Cardiovascular correlation genotype-phenotype of dopamine B-hydroxylase (DBH)

Introducción. Al convertir dopamina en norepinefrina la dopamina β-hidroxilasa (DβH) regula el tono dopaminérgico y adrenérgico. Los alelos rs1989787, rs1611115 y rs1108580 del gen DβH se asocian con actividad deficiente de la enzima y por eso evaluamos sus influencias sobre variables cardiovasculares clínicas, bioquímicas y farmacológicas. Métodos: A 44 voluntarios sanos con los haplotipos triple homocigoto nativo (CC/CC/AA), triple heterocigoto (CT/CT/AG), doble homocigoto mutado (TT/CC/GG) y homocigoto mutado para el rs1611115 (CC/TT/AA) les medimos presión arterial sistólica (PAS), presión arterial diastólica (PAD) y frecuencia cardíaca (FC) en posición decúbito, sentado, de pie, bajo estímulo de frío, con la maniobra de Valsalva y post ingestión de clonidina; adicionalmente medimos actividad enzimática por espectrofotometría y concentraciones séricas de dopamina y norepinefrina por ELISA. Resultados: Edad promedio 21,6±3,5 años, 54,5 porciento mujeres. No hubo diferencias de PAS, PAD y FC obtenidas bajo diferentes condiciones; sólo encontramos cambios significativos en los descensos de la PAS y la PAD postingestión de clonidina. No hubo relación de las variables clínicas, bioquímicas y farmacológicas mencionadas con los alelos rs1989787 y rs1108580, pero los portadores del alelo mutado T del rs1611115 sí tenían frecuencias cardíacas estadísticamente inferiores a los portadores del alelo nativo C. La actividad enzimática y las concentraciones de dopamina y norepinefrina no se correlacionaron con las variables cardiovasculares, pero se encontró correlación directa entre la actividad de la enzima y las concentraciones de norepinefrina. Conclusión: El polimorfismo -970C>T del gen DβH es el único asociado con menores frecuencias cardíacas en portadores del alelo mutado T.

Introduction. To the converting dopamine in norepinephrine the enzymedopamine β-hydroxylase (DβH) regulates dopaminergic and adrenergictone. The alleles rs1989787, rs1611115 and rs1108580 of the gene DβH areassociated with deficient activity of the enzyme and thus the influence ofthese SNPs in clinical, biochemical and pharmacological variables relatedwith the cardiovascular system was evaluated. Methods: 44 healthy volunteerswith homozygous triple native (CC/CC/AA), triple heterozygous (CT/CT/AG), double mutated homozygous (TT/CC/GG) and mutated homozygoushaplotypes for the rs1611115 (CC/TT/AA) were subjected to measurementsof systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate(HR) in the positions prone, sitting, standing, under cold stimulus, with theValsalva maneuver and after clonidine ingestion; additionally the enzymaticactivity of the DβH and the concentrations of dopamine and norepinephrinewere determined by spectrophotometry and ELISA, respectivelly. Results: Theaverage age of the group was 21.6±3.5 years, and 54.5% were women. Therewere no differences in SBP, DBP and HR obtained under different conditions;we only found significant changes in SBP and DBP decrease after clonidineingestion. There were no associations between clinical, biochemical andpharmacological variables with the rs1989787 and rs1108580 alleles, however,carriers of the mutant allele T of the rs1611115 had heart rates statisticallylower than the native C allele carriers. Neither enzymatic activity nor dopamineand norepinephrine levels were correlated with cardiovascular variables, but adirect correlation between enzyme activity and norepinephrine concentrationswas found. Conclusion: The-970C>T polymorphism of DβH gene is the onlyone associated with lower heart rates in carriers of the mutated allele T.

Association between polymorphism of dopamine-β-hydroxylase genes and violent behavior in adolescents / 重庆医学

Objective To explore the relationship between the rs1611115 and rs739398 polymorphism of dopamine-β-hydroxy-lase genes and violent behavior in adolescents .Methods Adolescent male offenders were divided into violent group (107 samples) and nonviolent group (107 samples) .Besides ,107 male adolescents in vocational school were served as normal controls .The rs1611115 and rs739398 polymorphisms of DβH gene were measured by used SNaPshot SNP technology in 321 samples .Chi-square test was used to compare the distribution difference between three groups of each gene type and the allele frequency distribution . Results There was statistically significant difference among violent group ,nonviolent group and normal control group in the geno-type and allele frequencies of DβH gene rs1611115 polymorphism(P0 .05) .There was no statistically significant difference among violent group ,nonvio-lent group and normal control group in the genotype and allele frequencies of rs739398 polymorphism(P>0 .05) .Conclusion No association was found between the rs739398 polymorphism of DβH gene and adolescents with violent behavior .But the rs1611115 polymorphism of DβH gene was related to adolescent criminals with violent and nonviolent behavior .

Electroacupuncture Analgesia Is Improved by Adenoviral Gene Transfer of Dopamine Beta-hydroxylase into the Hypothalamus of Rats

Electroacupuncture (EA) is a modified form of acupuncture that utilizes electrical stimulation. We previously showed that EA stimulated rats were divided into responders that were sensitive to EA and non-responders that were insensitive to EA based on the tail flick latency (TFL) test. The dopamine beta-hydroxylase (DBH) gene was more abundantly expressed in the hypothalamus of responder rats than non-responder rats. To determine whether overexpression of DBH gene expression in the hypothalamus modulate EA analgesia, we constructed a DBH encoding adenovirus and which was then injected into the hypothalamus of SD rats. Microinjection of DBH or control GFP virus into the hypothalamus had no changes on the basal pain threshold measured by a TFL test without EA treatment. However, the analgesic effect of EA was significantly enhanced from seven days after microinjection of the DBH virus, but not after injection of the control GFP virus. DBH expression was significantly higher in the hypothalamus of DBH virus injected rat than control GFP virus or PBS injected rats. Moreover, expression of the DBH gene did not affect the body core temperature, body weight, motor function or learning and memory ability. Although the functional role of DBH in the hypothalamus in the analgesic effect of EA remains unclear, our findings suggest that expression of the DBH gene in the hypothalamus promotes EA analgesia without obvious side-effects.

Xanthoangelol and 4-Hydroxyderricin Are the Major Active Principles of the Inhibitory Activities against Monoamine Oxidases on Angelica keiskei K

Monoamine oxidase inhibitors (MAOI) have been widely used as antidepressants. Recently, there has been renewed interest in MAO inhibitors. The activity-guided fractionation of extracts from Angelica keiskei Koidzumi (A. keiskei K.) led to the isolation of two prenylated chalcones, xanthoangelol and 4-hydroxyderricin and a flavonoid, cynaroside. These three isolated compounds are the major active ingredients of A. keiskei K. to inhibit the MAOs and DBH activities. Xanthoangelol is a nonselective MAO inhibitor, and a potent dopamine beta-hydroxylase (DBH) inhibitor. IC50 values of xanthoangelol to MAO-A and MAO-B were calculated to be 43.4 microM, and 43.9 microM. These values were very similar to iproniazid, which is a nonselective MAO inhibitor used as a drug against depression. The IC50 values of iproniazid were 37 microM, and 42.5 microM in our parallel examination. Moreover, IC50 value of xanthoangelol to DBH was calculated 0.52 microM. 4-Hydroxyderricin is a potent selective MAO-B inhibitor and also mildly inhibits DBH activity. The IC50 value of 4-hydroxyderricin to MAO-B was calculated to be 3.43 microM and this value was higher than that of deprenyl (0.046 microM) used as a positive control for selective MAO-B inhibitor in our test. Cynaroside is a most potent DBH inhibitor. The IC50 value of cynaroside to DBH was calculated at 0.0410 microM. Results of this study suggest that the two prenylated chalcones, xanthoangelol and 4-hydroxyderricin isolated from A. keiskei K., are expected for potent candidates for development of combined antidepressant drug. A. keiskei K. will be an excellent new bio-functional food material that has the combined antidepressant effect.

Effects of 6-hydroxydopamine and inhibitors for vesicular monoamine transporter on monoamine neurotransmitter and rate-limiting enzyme gene of them in PC12 cells / 中华神经科杂志

ObjectiveTo study the effects of 6-hydroxydopamine (6-OHDA),and inhibitors for vesicular monoamine transporter (VMAT) on 5-hydroxytryptamine (5-HT),norepinephrine (NE) and dopamine (DA) and the expressions of tryptophan hydroxylase (TpH) mRNA,dopamine-beta-hydroxylase (DβH) mRNA and tyrosine hydroxylase (TH) mRNA in PC12 cells.Methods The cell viability was determined using MTT assay, the density of 5-HT, NE and DA was detected using enzyme-linked immunosorbent assay,and the expressions of TpHmRNA,DβHmRNA and THmRNA were detected using RT-PCR in PC12 cells at different time points (0,12,24,36,48 h )after exposure to different concentrations of 6-OHDA(25,50,100,200 μmol/L),and VMAT inhibitors,reserpine (50,100,400,1600 nmol/L),which combined with 6-OHDA( 100 μmol/L).Results (1)The cell viability declined with the increasing concentration of 6-OHDA which showed time dependence.The cell viability in PC12 cell which treated with reserpine decreased significantly in the responding group.The density of 5-HT in PC12 cell did not decrease with the increasing concentration of 6-OHDA,but the change had the time dependence,and the density of 5-HT was lowest at 36 h.The density of NE decreased with the increasing concentration of 6-OHDA which showed time dependence. The density of DA in PC12 cell decreased with the increasing concentration of 6-OHDA,but the change did not have the time dependence.The density of 5-HT,NE and DA in PC12 cell which treated with reserpine decreased significantly in the responding group. (2) The expressions of TpHmRNA, DβHmRNA and THmRNA in PC12 cell decreased with the increasing concentration of 6-OHDA which showed time dependence.The expressions of TpHmRNA(0.006 ± 0.001,0.003 ± 0.000,0.003 ± 0.000,0.002 ± 0.000) ; DβHmRNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ±0.001,0.001 ± 0.000) and THm RNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ± 0.001,0.001 ± 0.000) in PC12 cell which treated with reserpine decreased significantly in the responding group(F =13.336,9.000,9.393,all P =0.000).Conclusions6-OHDA can decrease the cell viability in PC12 cell,reduce the density of 5-HT,NE and DA and decrease the expressions of TpHmRNA,DβHmRNA and THmRNA,and the effects have dose and time dependence.Reserpine can aggravate this damage.

Anti-stress effects of ginseng via down-regulation of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) gene expression in immobilization-stressed rats and PC12 cells

Catecholamines are among the first molecules that displayed a kind of response to prolonged or repeated stress. It is well established that long-term stress leads to the induction of catecholamine biosynthetic enzymes such as tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) in adrenal medulla. The aim of the present study was to evaluate the effects of ginseng on TH and DBH mRNA expression. Repeated (2 h daily, 14 days) immobilization stress resulted in a significant increase of TH and DBH mRNA levels in rat adrenal medulla. However, ginseng treatment reversed the stress-induced increase of TH and DBH mRNA expression in the immobilization-stressed rats. Nicotine as a ligand of the nicotinic acetylcholine receptor (nAChR) in adrenal medulla stimulates catecholamine secretion and activates TH and DBH gene expression. Nicotine treatment increased mRNA levels of TH and DBH by 3.3- and 3.1-fold in PC12 cells. The ginseng total saponin exhibited a significant reversal in the nicotine-induced increase of TH and DBH mRNA expression, decreasing the mRNA levels of TH and DBH by 57.2% and 48.9%, respectively in PC12 cells. In conclusion, immobilization stress induced catecholamine biosynthetic enzymes gene expression, while ginseng appeared to restore homeostasis via suppression of TH and DBH gene expression. In part, the regulatory activity in the TH and DBH gene expression of ginseng may account for the anti-stress action produced by ginseng.

Association of dopamine ?-hydroxylase polymorphism with attention deficit hyperactivity disorder in children / 北京大学学报(医学版)

Objective: To study the association of dopamine ?-hydroxylase gene 5′ flanking region polymorphism -1021 C→T with attention deficit hyperactivity disorder (ADHD) in Han children. Methods: ADHD was diagnosed according to the DSM-Ⅳ criteria . DNA was extracted and PCR was performed to examine the DBH-1021C→T polymorphism. HHRR was used to test the association of dopamine ?-hydroxylase gene with different subtypes of ADHD. Results: HHRR results showed this polymorphism had a tendency to be associated with the inattention subtype (P=0.067) and the combined subtype (P=0.076). The T allele was the protective factor of the inattentive subtype (P= 0.07), and the risk factor of the combined subtype (P=0.08). After dividing the 292 nuclear families according to sex, DBH-1021C→T polymorphism was found to be associated with the combined subtype (P=0.04) with the T allele as the risk factor (P=0.02). There were no positive findings among the girl nuclear families of ADHD. Conclusion: DBH gene is related to ADHD combined subtype and inattention subtype. The genetic basis of ADHD combined subtype and is inattention is different, and is influenced by sex factor.

Changed DBH expression of the neurons in lumbar sympathetic ganglia in response to Buerger's disease of lower limb / 重庆医科大学学报

Objective:To observe the expression of dopamme beta hydroxylase (DBH) in the lumbar sympathetic ganglia neurons of the patients with Buerger's disease of lower limb.Methods:The DBH expression of the neurons in lumbar sympathetic ganglia of 14 patients with Buerger's disease were observed by immunohistochemistry staining.11 patients without it served as the controls.Results:The DBH expression of the patients were reinforced that of control group.Conclusion:The enhanced expression of DBH may result in vasospasm,accelerating the development of Buerger's disease.