Mutación de protocadherina 19 (PC-DH19), en paciente con epilepsia refractaria / Mutation of protocadherin 19 (PCDH19), in patients with refractory epilepsy

Paciente de 4 años de edad, con epilepsia de difícil manejo, cuya etiología se atribuye a patología autoinmune y que finalmente se diagnostica una mutación de protocadherina (PCDH19). Se discute la fisiopatología, características clínicas, exámenes y los posibles tratamientos.

Four-year-old patient with intractable epilepsy, whose etiology is attributed to autoimmune pathology and who is eventually diagnosed with a protocadherin mutation (PCDH19). Pathophysiology, clinical characteristics, examinations and possible treatments are discussed.

Allopregnanolone suppresses mechanical allodynia and internalization of neurokinin-1 receptors at the spinal dorsal horn in a rat postoperative pain model

BACKGROUND: To identify a new strategy for postoperative pain management, we investigated the analgesic effects of allopregnanolone (Allo) in an incisional pain model, and also assessed its effects on the activities of the primary afferent fibers at the dorsal horn. METHODS: In experiment 1, 45 rats were assigned to Control, Allo small-dose (0.16 mg/kg), and Allo large-dose (1.6 mg/kg) groups (n = 15 in each). The weight bearing and mechanical withdrawal thresholds of the hind limb were measured before and at 2, 24, 48, and 168 h after Brennan's surgery. In experiment 2, 16 rats were assigned to Control and Allo (0.16 mg/kg) groups (n = 8 in each). The degree of spontaneous pain was measured using the grimace scale after the surgery. Activities of the primary afferent fibers in the spinal cord (L6) were evaluated using immunohistochemical staining. RESULTS: In experiment 1, the withdrawal threshold of the Allo small-dose group was significantly higher than that of the Control group at 2 h after surgery. Intergroup differences in weight bearing were not significant. In experiment 2, intergroup differences in the grimace scale scores were not significant. Substance P release in the Allo (0.16 mg/kg) group was significantly lower than that in the Control group. CONCLUSIONS: Systemic administration of Allo inhibited mechanical allodynia and activities of the primary afferent fibers at the dorsal horn in a rat postoperative pain model. Allo was proposed as a candidate for postoperative pain management.

Neurosteroid Levels in Patients with Obsessive-Compulsive Disorder

OBJECTIVE: Changes in serum neurosteroid levels have been reported in stress-related disorders such as anxiety and depression, but not in patients with obsessive-compulsive disorder (OCD). We thus investigated such changes in patients with OCD. METHODS: We compared the serum levels of progesterone, pregnanolone, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate (DHEA-S), cortisol and testosterone in 30 patients with OCD and 30 healthy controls. RESULTS: When male and female patients were evaluated together, DHEA and cortisol levels were significantly higher in patients with OCD than the control group. When the genders were evaluated separately, DHEA and cortisol levels were higher in female patients than the female controls. The increase in DHEA levels in female patients is likely an effect of the hypothalamic-pituitary-adrenal (HPA) axis. In contrast, cortisol levels in male patients were higher than the control group, while testosterone levels were lower. The increased cortisol and decreased testosterone levels in male patients likely involves the hypothalamic-pituitary-gonadal (HPG) axis. CONCLUSION: These findings suggest that neurosteroid levels in patients with OCD should be investigated together with the HPA and HPG axes in future studies.

Correlation between neurotransmitters and neurosteroids and premenstrual syndrome patients of Gan-yang ascending syndrome and Gan-qi stagnation syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the pathogenesis of premenstrual syndrome (PMS), and the correlation between anger and depression and PMS of Gan-yang ascending syndrome (GYAS) and Gan-qi stagnation syndrome (GQSS) by detecting the neuro-reproductive hormones of PMS patients of GYAS and GOSS, thus providing theoretical reliance for diagnostic standards for clinical normative PMS.</p><p><b>METHODS</b>Using techniques such as HPLC, HPLC-MC, ELISA, and radioimmunoassay (RIA), levels of serum sex hormones (follicle stimulating hormone, luteinizing hormone, estradiol, progesterone, testosterone, and prolactin), plasma neurotransmitters (gamma-aminobutyric acid, beta-endorphin, glutamic acid, dopamine, 5-HT, adrenaline, and noradrenaline), neurosteroids (allopregnanolone, pregnenolone, and dehydroepiandrosterone) in the follicular phase and the luteal phase of PMS patients of GYAS (30 cases) and GQSS (30 cases) were detected, and compared with the healthy control group (30 cases).</p><p><b>RESULTS</b>There was no statistical difference in either index of the follicular phase among the 3 groups. Compared with the healthy control group, the testosterone level in PMS patients of GYAS in the luteal phase showed increasing tendency (P > 0.05). The levels of dopamine and 5-HT of PMS patients of GYAS in the luteal phase were higher and the gamma-aminobutyric acid level was lower than those of the healthy control group (all P < 0.05). The levels of adrenaline and noradrenaline of PMS patients of GYAS and GQSS in the luteal phase were higher than those of the healthy control group (all P < 0.05). The levels of allopregnanolone and pregnenolone of PMS patients of GYAS and GQSS in the luteal phase were lower, and the dehydroepiandrosterone level was higher than those of the healthy control group (all P < 0.05). The ratios of dehydroepiandrosterone/allopregnanolone and dehydroepiandrosterone/pregnenolone of PMS patients of GYAS and GQSS in the luteal phase were higher than those of the healthy control group (P < 0.05).</p><p><b>CONCLUSION</b>The decreased levels of pregnenolone and allopregnanolone, increased dehydroepiandrosterone levels, and increased ratios of dehydroepiandrosterone/allopregnanolone and dehydroepiandrosterone/pregnenolone might be one of biological factors for anger and depression in PMS patients of GYAS and GQSS.</p>

Effects of neuroactive steroid allopregnanolone on the damage of cortical neurons / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the protective mechanism of neuroactive steroid allopregnanolone on N-methyl-D-aspartate (NMDA) induced toxicity in primary mouse cortical neurons.</p><p><b>METHODS</b>Primary cultured mouse cortical neurons were subjected to allopregnanolone, the expression of beta-aminobutyric acid receptor beta2 subunit (beta2-GABA-R) mRNAs was detected by RT-PCR and Akt phosphorylation was assayed by Western blot using Akt-phosphoserine 473-specific antibody. After the cultured mouse cortical neurons were pretreated with or without allopregnanolone prior to treatment with NMDA , DNA isolated was analyzed by agarose gel electrophoresis and proteins collected were analyzed by Western blot with anti-cleaved-PARP, anti-cleaved caspase-3, and anti-cleaved caspase-9 antibodies.</p><p><b>RESULTS</b>When cultured mouse cortical neurons were exposed to allopregnanolone both the expression of beta2-GABA-R mRNAs and Akt phosphorylation increased. Allopregnanolone inhibited the NMDA-induced apoptosis and decreased the level of active-PARP, active-caspase-3 and active-caspase-9 notably at a final concentration of 5 x 10(6) mol/L.</p><p><b>CONCLUSION</b>Pretreatment with allopregnanolone may be neuroprotective on NMDA-induced neuronal cells apoptosis by increasing beta2-GABA-R expression and Akt phosphorylation.</p>

[Comparison of the effects of progesterone, allopregnanolone and gender on suppressing edema formation after traumatic brain injury in rats]

Brain edema is one of the most important causes of death within the first few days following head trauma. In this study we investigated the role of gender as well as the effects of progesterone and allopregnanolone one hour after diffuse traumatic brain injury on edema formation in rats. This interventional-experimental study was performed on 12 groups of female and male rats. They were divided into 12 groups as follows: 1 and 2: intact female and male rats, 3 and 4: trauma male and female rats, 5: vehicle of progesterone [benzyl alcohol with sesame oil], 6: sham [ovariectomized female rats: ovx], 7: sham [no ovx], 8: sham[male], 9 and 10: low dose [4mg/kg] and high dose [8mg/kg] of progesterone, 11: allopregnanolone and 12: vehicle of allopregnanolone [water]. Hormones were injected i. p one hour after diffuse traumatic brain injury through Marmarou model. The results showed a significant increase of 5.32 times in Evans blue and 2.42% in water content in trauma male group comparing to control groups, while in female rats the difference was significant just for Evans blue [4.68 times]. Evans blue and water content were also significantly greater in traumatic males than female rats [1.57 times and 2.04% respectively]. After injection of low and high doses of progesterone, there was a significant decrease in water content [2.21% and 2.30%] and Evans blue content [2.55 and 2.98 times]. Allopregnanolone significantly decreased these parameters [2.36% and 1.82 times respectively]. Moreover, the injection of progesterone in both low and high doses increased the serum progesterone of female ovarectomized rats as compared to vehicle group. Based on these results, it can be concluded that the rate of edema formation in traumatic male rats is higher compared to traumatic female rats. Moreover, both progesterone and allopregnanolone decrease edema formation in ovariectomized female rats

Influence of epipregnanolone on the modulation of rapid tolerance to ethanol by neurosteroids

OBJECTIVE: The objective of the present study was to investigate the effect of epipregnanolone on the influence of neurosteroids on the development of rapid tolerance to the motor impairing and hypothermic effects of ethanol. METHOD: Experiment 1: on Day 1 groups of mice were pretreated with saline or with epipregnanolone. After 30 min each group was further divided in subgroups that received ethanol or saline. Thirty, 60 and 90 min after the injections the animals were tested on the rota-rod or the body temperature was measured. On Day 2 all groups received ethanol and a similar procedure was followed to evaluate rapid tolerance. Experiment 2 and 3: On Day 1 groups of mice were treated with epipregnanolone and after 15 min each group was divided into three groups in order to receive pregnenolone sulfate, dehydroepiandrosterone sulfate or saline. Thirty minutes later, each group was further divided into two subgroups in order to receive ethanol or saline, respectively, and 30, 60 and 90 min later the animals were tested as in the experiment 1. On Day 2 all groups received ethanol and a similar procedure was followed to evaluate rapid tolerance. RESULTS: Pretreatment with epipregnanolone (0.10-0.30 mg/kg) significantly blocked the development of tolerance to the motor impairing and hypothermic effects induced by ethanol in mice. Considering tolerance to ethanol-induced motor impairment, epipregnanolone (0.15 mg/kg) reversed the stimulatory action of dehydroepiandrosterone sulfate (0.15 mg/kg), but did not affect the actions of pregnenolone sulfate (0.08 mg/kg). Moreover, epipregnanolone prevented the inhibitory action of allotetrahydrodeoxycorticosterone (0.10 mg/kg). In relation to ethanol-induced hypothermia, the results showed that pretreatment with epipregnanolone (0.30 mg/kg) significantly prevented the stimulatory action of dehydroepiandrosterone sulfate and pregnenolone sulfate, as well as the inhibitory action of...

OBJETIVO: O objetivo do presente estudo foi o de investigar o efeito da epipregnanolona sobre a influência de neuroesteróides no desenvolvimento da tolerância rápida aos efeitos de incoordenação motora e hipotermia induzidos pelo etanol. MÉTODO: Experimento 1: no Dia 1, grupos de camundongos foram pré-tratados com salina ou com epipregnanolona. Após 30 min, cada grupo foi subdividido recebendo etanol ou salina. Aos 30, 60 e 90 min após as injeções, os animais foram testados no rota-rod ou a temperatura corporal foi avaliada. No Dia 2, todos os grupos receberam etanol e um procedimento similar foi seguido para avaliar a tolerância rápida. O pré-tratamento com a epipregnanolona (0,10-0,30 mg/kg) bloqueou significantemente o desenvolvimento da tolerância aos efeitos de incoordenação motora e hipotermia induzidos pelo etanol em camundongos. Experimento 2 e 3: no Dia 1, grupos de animais foram tratados com epipregnanolona e, após 15 min, cada grupo foi dividido em três grupos para receber sulfato de pregnanolona, sulfato de dehidroepiandrosterona ou salina. Após 30 min, cada grupo foi dividido em dois subgrupos para receber etanol ou salina, respectivamente, e após 30, 60 e 90 min os animais foram testados como no experimento 1. No Dia 2, todos os grupos receberam etanol e 30 min após foram testados como mencionado no experimento 1. RESULTADOS: Considerando a tolerância ao prejuízo motor induzido pelo etanol, a epipregnanolona (0,15 mg/kg) bloqueou a ação estimulatória do sulfato de dehidroepiandrosterona (0,15 mg/kg), mas não afetou a ação do sulfato de pregnanolona (0,08 mg/kg). Entretanto, a epipregnanolona bloqueou a ação inibitória da alotetrahidrodeoxicorticosterona (0,10 mg/kg). Em relação à hipotermia induzida pelo etanol, os resultados demonstraram que o pré-tratamento com epipregnanolona (0,30 mg/kg) bloqueou significantemente a ação estimulatória do sulfato de dehidroepiandrosterona e do sulfato de pregnanolona, bem como a ação...

Effects of morphine dependence on the levels of neurosteroids in rat brain / 药学学报

<p><b>AIM</b>To establish the rat model of morphine-induced conditioned place preference (CPP) and to investigate the effects of morphine psychical dependence on the levels of neurosteroids in rat brain.</p><p><b>METHODS</b>Rats were ip administered morphine 5 mg x kg(-1) for 10 days to induce CPP in morphine group. The concentrations of dehydroepiandrosterone (DHEA), pregnenolone (PREG), allopregnanolone (AP), dehydroepiandrosterone sulfate (DS) and pregnenolone sulfate (PS) in nucleus accumbens (Nac), hypothalamus (Ht), amygdale (A) and plasma of rats were determined with liquid chromatography-negative atmospheric pressure ionization mass spectrometry (LC-MS).</p><p><b>RESULTS</b>Trained with morphine for 10 days resulted in the acquisition of CPP in morphine group with the time that the rats spent in drug-pairing room was longer than that of control group. Compared with control group, morphine treatment could significantly decrease the contents of DHEA in Nac and plasma, decrease that of PREG in Ht.</p><p><b>CONCLUSION</b>Morphine could induce the CPP in rats and affected the contents of some neurosteroids in rat brain, which suggests that endogenous neurosteroids might he related to the development of morphine dependence.</p>

Pregnanolone effects on the blood pressure of stress-induced hypertension in rats / 生理学报

This study was conducted to determine the effect of pregnanolone (PGN) on blood pressure of a rat model of stress-induced hypertension (SIH). This model was established by applying electric shock to animal feet together with noise. PGN was administered intraperitoneally at 0.24 mg/kg.d(-1) and blood pressure, angiotensin II (Ang II) levels, and the expression of Fos-like protein immunoreactive (FLI) neurons in brain areas were determined. Rats were randomly divided into five groups: (1) control, (2) stressed for 1 h, (3) stressed for 1 h after PGN pretreatment, (4) stressed for a 2 h session, twice a day, for 15 d, and (5) stressed for a 2 h session after PGN pretreatment, twice a day, for 15 d. The results showed that increased systolic pressure of tail artery caused by a 15-d stress treatment was significantly reduced by PGN pretreatment (P<0.001). Ang II levels, measured by radioactive immunoreactivity, were significantly elevated (P<0.001) after the rats were stressed for 1 h or 15 d, the Ang II level was significantly reduced by PGN treatment in both 1 h and 15 d stress groups (P<0.05). Only a small number of FLI neurons were found in the brain areas of the control group, 15 d stress group, and 15 d stress with PGN group. In the 1 h stress group, more FLI neurons were found in the lateral habenular nucleus, the medial habenular nucleus, the paraventricular nucleus, the central nucleus of amgydaloid and the lateral hypothalamus compared with the control group. PGN pretreatment significantly prevented the increase in the number of FLI neurons. These results indicate that PGN pretreatment prevents elevation of tail artery systolic pressure in SIH rats and that this effect of PGN may be mediated through reducing Ang II level and inhibiting the activity of cardiovascular center involved in stress.

The Effect of Repeated Stress on the Modulation of Neuroactive Steroids at the GABAA-Benzodiazepine Receptor Complex in Rats / 신경정신의학

OBJECTIVES: Pregnanolone is a potent positive modulator of the gamma-aminobutyric acid(GABA) response that enhances the binding of [3H]flunitrazepam to the GABA A receptor. Recently, it was reported that chronic treatment with pregnanolone uncouples allosteric interactions between steroid and benzodiazepine recognition sites. The present study was designed to assess the effect of repeated stress on the modulation of neuroactive steroids on the GABA A receptor. METHODS: The effect of steroids on the ligands binding to GABA A receptor was investigated using cerebral cortices of unstressed and repeatedly immobilized rats. Male Sprague-Dawley rats, weighing 200-250g were forced to suffer an immobilization stress for 2 hours. RESULTS: Pregnanolone enhanced the binding of [3H]flunitrazepam to GABA A receptor in both of unstressed and repeatedly stressed rats. However, repeatedly stressed rats showed significantly higher values in EC50 and lower values in E max of enhancement binding of [3H]flunitrazepam than those of unstressed rats. CONCLUSIONS: From these findings, it can be concluded that repeated stress reduced the positive modulation of neuroactive steroid on the GABA A-receptor complex.

The Effect of Neuroactive Steroids on the GAVA-Benzodiazepine Receptor Complex / 대한정신약물학회지

Pregnolone[5beta-pregnan-3alpha-ol-one(5beta3alpha)] and allopregnanolone [(5alpha-pregnan-3alpha-ol-20-one(5alpha3alpha))] are neuroactive steroids that are reduced metabolites of progesterone. It was reported that Neuroactive steroids may have anxiolytic and anticonvulsant action similar to benzodiazepines and barbiturates. Therefore, the present study was designed to assess the interaction of steroids with GABAA-benzodiazepine receptor complex. The effect of steroids on the ligands binding to GABAA receptor complex was investigated using rat cortices. 5beta3alpha and 5alpha3alpha enhanced the binding of [3H] flunitrazepam to GABAA receptor, but testosterone, progesterone and dexamethasone did not. GABA also showed the enhancement of [3H] flunitrazepam binding, but did not show the additive effect. Unlike to GABA, 5beta3alpha and 5alpha3alpha did not affect on the [3H] muscimol binding to rat cortices. From these findings, it can be concluded that Neuroactive steroids are potent positive modulators of the GABA A receptor, and do not act at GABA binding site.

Anaesthetic steroids--a review.

Steroids produce anaesthesia besides producing the well known metabolic and hormonal effects. A number of anaesthetic steroids have been synthesized and tried clinically. Hydroxydione, althesin, minaxolone and pregnanolone are among those studied in detail. They act through GABAergic mechanism and are known to be advantageous over barbiturates.