The traditionally used oriental herbal medicine Moutan Cortex Radicis [MCR; Paeonia Suffruticosa Andrews (Paeoniaceae)] exerts anti-inflammatory, anti-spasmodic, and analgesic effects. In the present study, we investigated the therapeutic effects of differently fractioned MCR extracts in a 6-hydroxydopamine (OHDA)-induced Parkinson's disease model and neuro-blastoma B65 cells. Ethanol-extracted MCR was fractionated by n-hexane, butanol, and distilled water. Adult Sprague-Dawley rats were treated first with 20 μg of 6-OHDA, followed by three MCR extract fractions (100 or 200 mg·kg) for 14 consecutive days. In the behavioral rotation experiment, the MCR extract-treated groups showed significantly decreased number of net turns compared with the 6-OHDA control group. The three fractions also significantly inhibited the reduction in tyrosine hydroxylase-positive cells in the substantia nigra pars compacta following 6-OHDA neurotoxicity. Western blotting analysis revealed significantly reduced tyrosine hydroxylase expression in the substantia nigra pars compacta in the 6-OHDA-treated group, which was significantly inhibited by the n-hexane or distilled water fractions of MCR. B65 cells were exposed to the extract fractions for 24 h prior to addition of 6-OHDA for 30 min; treatment with n-hexane or distilled water fractions of MCR reduced apoptotic cell death induced by 6-OHDA neurotoxicity and inhibited nitric oxide production and neuronal nitric oxide synthase expression. These results showed that n-hexane- and distilled water-fractioned MCR extracts inhibited 6-OHDA-induced neurotoxicity by suppressing nitric oxide production and neuronal nitric oxide synthase activity, suggesting that MCR extracts could serve as a novel candidate treatment for the patients with Parkinson's disease.
Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Antiparkinson Agents ; pharmacology ; therapeutic use ; Cell Death ; drug effects ; Cell Line ; Disease Models, Animal ; Drugs, Chinese Herbal ; chemistry ; Neurons ; pathology ; Nitric Oxide ; analysis ; Nitric Oxide Synthase Type I ; biosynthesis ; Oxidopamine ; toxicity ; Paeonia ; chemistry ; Parkinsonian Disorders ; chemically induced ; drug therapy ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Plants, Medicinal ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects ; enzymology ; Tyrosine 3-Monooxygenase ; genetics ; metabolism

Calcium Channels, L-Type ; genetics ; metabolism ; Esophageal Achalasia ; genetics ; metabolism ; Esophagus ; metabolism ; Humans ; Nitric Oxide Synthase ; metabolism ; Nitric Oxide Synthase Type I ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, Calcium-Sensing ; genetics ; metabolism

OBJECTIVEThe study was aimed to investigate the inhibitory effect and mechanism of astragaloside IV (ASI) on the activation of microglial cells.

METHODAfter pre-incubated with ASI for 2 h, microglial cells BV-2 were stimulated with interferon-γ (IFN-γ) for 1. 5 h and 24 h, respectively. Secretion of nitric oxide (NO) in the medium was measured by Griess method. Production of tumor necrosis factor alpha (TNF-α) was detected by ELISA approach. Cellular gene expressions of CD11b, TNF-α, interleukin 1β (IL-1β) and induced nitric oxide synthase (iNOS) were examined by quantitative-PCR analysis. Total and phosphorylation of STAT1, IκB and NF-κB was analyzed by Western blot method.

RESULTASI could significantly inhibit the increased secretion of TNF-α and NO from BV-2 cells upon IFN-γ stimulation (P < 0.001). Further study showed that ASI significantly down-regulated gene expression of IL-1β and TNF-α (P < 0.01, P < 0.05) and exhibited a trend to reduce that of iNOS. IFN-γ and ASI have no obvious effect on gene expression of CD11b. Moreover, ASI inhibited the phosphorylation of STAT1, IκB and NF-κB elicited by IFN-γ stimulation.

CONCLUSIONASI could restrain microglial activation through interfering STAT1/IκB/NF-κB signaling pathway, reducing gene expres- sion of IL-1β and TNF-α, and thus inhibiting the production of proinflammatory mediators such as NO and TNF-α.

Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; I-kappa B Proteins ; genetics ; metabolism ; Interferon-gamma ; genetics ; metabolism ; Mice ; NF-kappa B ; genetics ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; STAT1 Transcription Factor ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology

OBJECTIVETo explore the mechanism of electroacupuncture (EA) in the treatment of post-traumatic stress disorder (PTSD).

METHODSThirty male Sprague-Dawley rats were randomly divided into a normal group, a model group and an electroacupuncture group. The single prolonged stress (SPS) method was used to set up the PTSD models in latter two groups. After SPS Stimulation, EA group was treated with 2Hz electroacupuncture at Baihui (GV 20) and Zusanli (ST 36) for 30 min, once a day for a week. Reverse transcriptase polymerase chain reaction (RT-PCR) and immuno-histochemistry were used to detect the mRNA and protein expression of nNOS in the hippocampus of rats in the each group.

RESULTS(1) The nNOS mRNA expression in hippocampus in model group was higher than that in normal group (P < 0.05). But the expression in EA group was lower significantly than that in model group (P < 0.05). (2) The nNOS protein expression in hippocampus CA1 and CA3 in model group was higher than that in normal group (P < 0.05). But after electroacupuncture treatment, its expression in EA group was lower significantly than that in model group (P < 0.05). The nNOS protein expression in hippocampal CA2 had no difference among all three groups.

CONCLUSIONThe elevated nNOS expression in hippocampus may be involved in the pathological process of PTSD. Electroacupuncture play a down-regulation effects in the hippocampal nNOS expression, which may be one mechanism of electroacupuncture for treatment of PTSD.

Animals ; Electroacupuncture ; Hippocampus ; enzymology ; Humans ; Male ; Nitric Oxide Synthase Type I ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stress Disorders, Post-Traumatic ; enzymology ; genetics ; therapy

OBJECTIVETo investigate the distribution of GAD67 and the co-localization with bNOS in the main olfactory bulb of GAD67-GFP knock-in mouse.

METHODSPolymerase chain reaction was applied to identify the genotype of GAD67-GFP knock-in mouse, the animals were sacrificed and frozen sections of olfactory bulb were prepared. The Nissl-staining was performed to show an framework of the neuron in the olfactory bulb. The distribution of GAD67 and co-localization with bNOS were detected by immunofluorescence technique.

RESULTSThe proportion of GAD67-positive cells among DAPI-positive cells were (42.98 ± 0.92)% in glomerular layer, (23.64 ± 0.84)% in mitral cell layer and (77.75 ± 0.84)% in granule cell layer; the bNOS-positive cells mainly existed in glomerular layer and mitral cell layer, very few in granule cell layer. No co-localization of GAD67 and bNOS in granule cell layer and mitral cell layer was found, but there was dispersed distribution in glomerular layer.

CONCLUSIONGAD67-positive neurons mainly appear in glomerular layer and granule cell layer, and the bNOS is mostly expressed in glomerular layer and mitral cell layer; while the co-localization of GAD67 and bNOS only occurs in glomerular layer of olfactory bulb.

Animals ; Gene Knock-In Techniques ; Glutamate Decarboxylase ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Mice, Transgenic ; Neurons ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; Olfactory Bulb ; metabolism ; Tissue Distribution

OBJECTIVETo assess the association between nitric oxide synthase 1 (NOS1) gene polymorphisms and schizophrenia.

METHODSTwenty eight tag single nucleotide polymorphisms (SNPs) of NOS1 in 382 schizophrenic patients and 448 healthy individuals sampled from Chinese Han population were analyzed by a Illumina GoldenGate Genotyping Assay.

RESULTSOne SNP (rs1520811) was found to be associated with schizophrenia, which however becomes negative after Bonferroni correction (P>0.05). Further analysis has failed to identify any association between particular haplotypes and the disease.

CONCLUSIONOur results did not support a significant association between NOS1 gene polymorphisms and schizophrenia.

Adult ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Male ; Nitric Oxide Synthase Type I ; genetics ; Polymorphism, Single Nucleotide ; Schizophrenia ; enzymology ; genetics ; Young Adult

This study is to investigate the effects on the expression of iNOS and production of NO in the osteogenic differentiation process of rat bone marrow stromal cells (rBMSCs) by icariside II. rBMSCs were cultured by adherence screening method. When the culture dishes were covered with 80% cells, the osteogenic induced cultures were adopted. Icariside II was supplemented into the culture at 1 x 10(-5) mol x L(-1). The activity of iNOS, content of NO and osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-FALP and mineralized bone nodules were compared among the icariside II-supplemented group, L-NMAE group, icariside II + L-NAME group and the control. Total RNA was isolated and the gene expression of iNOS, Osterix and Runx-2 was investigated by real-time PCR. Total protein was also isolated and the secretion of iNOS and collagen I was examined by Western blotting. Icariside II can significantly improved ALP activity, CFU-FALP amount and mineralized nodules. Besides, the mRNA level of factors related to the osteogenic differentiation includes Osterix and Runx-2 also enhanced. The secretion of collagen I also promoted significantly. But all of these effects can be inhibited by L-NAME which can specifically inhibit the activity of iNOS. Icariside II enhances the osteogenic differentiation of rBMSCs significantly, but if the activity of iNOS was blocked by L-NAME, the osteogenic differentiation markers decrease accompanied with iNOS and NO decrease, suggesting that icariside II stimulates the osteogenic differentiation via enhancing the activity of iNOS and promoting the generation of NO.
Alkaline Phosphatase ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Osteogenesis ; drug effects ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Transcription Factors ; genetics ; metabolism

Animals ; Fatigue ; enzymology ; Male ; Mammillary Bodies ; metabolism ; Nitric Oxide Synthase Type I ; genetics ; metabolism ; Physical Exertion ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Stress, Physiological ; physiology

OBJECTIVETo study the expression of heme oxygenase (HO) in the corpus cavernosum of castrated rats and to investigate its role in androgen deficiency-mediated erectile dysfunction.

METHODSForty 10-week old SD rats were randomly divided into Groups A (2-week sham operation), B (4-week sham operation), C (2-week castrated) and D (4-week castrated). The serum testosterone level was determined, and the expressions of HO and nNOS in the corpus cavernosum of the castrated rats were detected by immunohistochemical staining and RT-PCR 2 and 4 weeks after the operation.

RESULTSCompared with the sham operation groups, the castrated rats showed a significant reduction in the level of serum testosterone, (A vs. C: [283.222 +/- 117.171] ng/dl vs. [7.117 +/- 3.700] ng/ dl; B vs. D: [289.280 +/- 87.413] ng/dl vs [48.826 +/- 19.477] ng/dl) (P < 0.01), the expressions of HO-1 and HO-2 proteins (P < 0.01) and the expressions of HO-1, HO-2 and nNOS mRNA (P < 0.01).

CONCLUSIONAndrogen can partly regulate the erectile function of the penis via the HO-CO system.

Animals ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Orchiectomy ; Penis ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; blood

OBJECTIVETo study the effect of the Chinese compound prescription Ginkgo biloba Pingchan Recipe (GBPR) on experimental Parkinson disease (PD) in mice and explore the possible mechanism.

METHODSMale C57/BL6J mice were divided into normal control, PD model and treatment groups. PD model was established by intraperitoneal injection with 1-methl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP) in the mice, and in the treatment group, GBPR was administered intragastrically after the injection. The mice were sacrificed 14 and 28 days later, and using in situ hybridization with Digoxin-labeled nNOS cDNA oligonucleotide probe, neuronal nitric oxide synthase (nNOS) mRNA was detected in the striatum and substantia nigra in the brain of mice.

RESULTSnNOS mRNA expression was detected in the striatum and substantia nigra of the PD model mice, and GBPR treatment significantly reduced its expressions.

CONCLUSIONGBPR has obvious inhibitory effect against the neurotoxicity of NO probably by producing an anti-oxiditive effect through decreasing nNOS synthesis in the brain.

Animals ; Brain ; drug effects ; enzymology ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Ginkgo biloba ; chemistry ; Male ; Mice ; Mice, Inbred C57BL ; Neostriatum ; drug effects ; metabolism ; Nitric Oxide Synthase Type I ; genetics ; Parkinson Disease ; enzymology ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Substantia Nigra ; drug effects ; metabolism