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.
Anxiety ; Axis, Cervical Vertebra ; Dehydroepiandrosterone ; Depression ; Female ; Humans ; Hydrocortisone ; Male ; Obsessive-Compulsive Disorder* ; Pregnanolone ; Progesterone ; Testosterone

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.
Animals ; Benzodiazepines ; Cerebral Cortex ; gamma-Aminobutyric Acid ; Humans ; Immobilization ; Ligands ; Male ; Pregnanolone ; Rats* ; Rats, Sprague-Dawley ; Receptors, GABA-A ; Steroids*

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.
Animals ; Barbiturates ; Benzodiazepines ; Binding Sites ; Dexamethasone ; Flunitrazepam ; gamma-Aminobutyric Acid ; Ligands ; Muscimol ; Pregnanolone ; Progesterone ; Rats ; Receptors, GABA-A ; Steroids* ; Testosterone

Animals ; Habenula ; cytology ; drug effects ; Male ; Microelectrodes ; Neurons ; drug effects ; physiology ; Pain ; Pregnanolone ; pharmacology ; Rats ; Rats, Wistar

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.
Animals ; Extremities ; Hyperalgesia ; Pain, Postoperative ; Pregnanolone ; Rats ; Receptors, Neurokinin-1 ; Spinal Cord ; Spinal Cord Dorsal Horn ; Substance P ; Weight-Bearing

OBJECTIVETo 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.

METHODSUsing 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).

RESULTSThere 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).

CONCLUSIONThe 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.

Adult ; Case-Control Studies ; Dehydroepiandrosterone ; blood ; Estradiol ; blood ; Female ; Follicular Phase ; blood ; Humans ; Luteal Phase ; blood ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Neurotransmitter Agents ; Pregnanolone ; blood ; Pregnenolone ; blood ; Premenstrual Syndrome ; blood ; diagnosis ; Progesterone ; blood ; Young Adult

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.
Angiotensin II ; metabolism ; Animals ; Blood Pressure ; drug effects ; Brain ; metabolism ; Electric Stimulation ; Hypertension ; etiology ; physiopathology ; Male ; Pregnanolone ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Random Allocation ; Rats ; Rats, Wistar ; Stress, Physiological ; complications

AIMTo 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.

METHODSRats 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).

RESULTSTrained 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.

CONCLUSIONMorphine 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.

Amygdala ; metabolism ; Animals ; Brain ; metabolism ; Conditioning, Operant ; physiology ; Dehydroepiandrosterone ; blood ; metabolism ; Dehydroepiandrosterone Sulfate ; blood ; metabolism ; Hypothalamus ; metabolism ; Male ; Morphine Dependence ; metabolism ; Nucleus Accumbens ; metabolism ; Pregnanolone ; blood ; metabolism ; Pregnenolone ; blood ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the protective mechanism of neuroactive steroid allopregnanolone on N-methyl-D-aspartate (NMDA) induced toxicity in primary mouse cortical neurons.

METHODSPrimary 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.

RESULTSWhen 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.

CONCLUSIONPretreatment with allopregnanolone may be neuroprotective on NMDA-induced neuronal cells apoptosis by increasing beta2-GABA-R expression and Akt phosphorylation.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cerebral Cortex ; cytology ; Mice ; N-Methylaspartate ; antagonists & inhibitors ; toxicity ; Neurons ; cytology ; Neuroprotective Agents ; pharmacology ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Pregnanolone ; pharmacology ; Primary Cell Culture ; RNA, Messenger ; genetics ; metabolism ; Receptors, GABA-B ; genetics ; metabolism