In order to further investigate the mechanisms of action of berberine (Ber), we assessed the effects of Ber on the mRNA expression of nitric oxide synthases (NOS) in rat corpus cavernosum. After incubation with Ber for 1 or 3 h respectively, the levels of NOS mRNA were examined by reverse transcription polymerase chain reaction (RT-PCR). Our results showed that there were iNOS and eNOS mRNA expressions in rat corpus cavernosum. Ber enhanced eNOS mRNA expression in rat penis, but exhibited no effect on the expression of iNOS mRNA (P > 0.05). The present study indicated that the relaxation of Ber involved the NO-cGMP signal transduction pathway. The enhancing effect of Ber on eNOS mRNA expression might associated with its relaxation of corpus cavernosum.
Berberine/*pharmacology ; Connective Tissue/physiopathology ; Nitric Oxide Synthase/*biosynthesis ; Nitric Oxide Synthase/genetics ; Nitric Oxide Synthase Type I/biosynthesis ; Nitric Oxide Synthase Type I/genetics ; Nitric Oxide Synthase Type III/biosynthesis ; Nitric Oxide Synthase Type III/genetics ; Penile Erection/*physiology ; Penis/*metabolism ; Penis/physiology ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics

OBJECTIVESTo investigate the effect of aging on the expression of nitric oxide synthase I (NOS I) and the activity of nitric oxide synthase in the rat penis.

METHODSThirty male rats from three age groups(adult, old and senescent) were investigated: 1. The expressions of NOS I protein and mRNA in the penis were detected by Western blot and RT-PCR, respectively. 2. NOS activity in the penis was detected with ultraviolet spectrophotometry.

RESULTSIn rats of the old and the senescent groups, the expression of NOS I protein decreased significantly as compared to that of the adult group. NOS I mRNA expression was well related to its protein expression. NOS activity had no statistical difference between the adult group and the old group, but it reduced significantly in the senescent group as compared to that of the adult group(P < 0.01).

CONCLUSIONSIt is maybe one of the main mechanisms of erectile dysfunction in the aging male that the aging causes the decreases of NOS I protein and mRNA expression and NOS activity.

Aging ; metabolism ; Animals ; Erectile Dysfunction ; enzymology ; Male ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type I ; Penis ; enzymology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate protein and mRNA expression of nitric oxide synthase (NOS) isoforms I and III in rat penile tissues.

METHODSPenile tissues from male SD rats were investigated. The protein expression of NOS I and III in rat penis was confirmed by immunoprecipitation and Western blot. Gene expression of both NOS I and III was identified by reverse transcription-polymerase chain reaction.

RESULTSThe mRNA and protein products of NOS I and III are both expressed in normal rat penile spongium tissues. But, NOS III mRNA and protein products are more highly expressed than those of NOS.

CONCLUSIONSThese findings support the possibility that penile erection is regulated by different NOS isoforms in rat penile tissues.

Animals ; Blotting, Western ; Male ; Nitric Oxide Synthase ; analysis ; genetics ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type III ; Penis ; enzymology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate gene expression of three nitric oxide synthase isozymes in injured spinal cord tissue.

METHODSThirty-six adult SD rats were randomly divided into six groups: a normal group and five injury groups, with six per each group. Animals in the injury groups were sacrificed at 2, 6, 12, 24, 48 h after injury. A compression injury model on the spinal cord was made according to Nystrom B et al and gene expression of the three NOS isozymes were examined by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSGene expression of nNOS and eNOS were detectable in the normal group and were up-regulated quickly after injury, reaching a maximum at 6 h: (0.633 +/- 0.012) and (1.236 +/- 0.207). Gene expression of iNOS was detectable only in the injury groups and it was gradually up-regulated after injury, reaching a maximum at 24 h: (1.043 +/- 0.049).

CONCLUSIONInjury to the spinal cord leads to early up-regulation of cNOS and late up-regulation of iNOS. Different NOS isozymes may play different roles in secondary spinal cord injury.

Animals ; Female ; Gene Expression Regulation, Enzymologic ; Male ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type II ; Nitric Oxide Synthase Type III ; RNA ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Spinal Cord ; enzymology ; pathology ; Spinal Cord Injuries ; enzymology ; genetics

OBJECTIVETo investigate the gene and protein expressions of three isoforms of nitric oxide synthase (NOS) and gene expression of Caspase-3, and effect of dexamethasone on them in neonatal rats with lipopolysaccharide (LPS)-induced endotoxemic brain damage.

METHODSExpressions of the three isoforms of NOS and caspase-3 mRNA in the brain were investigated by RT-PCR in postnatal 7-day Wistar rats with acute endotoxemia by intraperitoneal administration of LPS. Regional distributions of NOSs were examined by immunohistochemical technique.

RESULTSnNOS and Caspase-3 mRNA were obviously detected. eNOS mRNA was faintly expressed, but iNOS mRNA was undetectable in the control rat brain. The expressions of NOS mRNA of three isoforms were weak 2 h after LPS (5 mg/mg) delivery, peaked at 6 h, and thereafter, reduced gradually up to 24 h. The expression intensity was in the order of nNOS> iNOS> eNOS. Widespread nNOS, scattered eNOS distribution and negative iNOS were identified in the control rat brain and all isoforms of NOS could be induced by LPS which reached the apex at 24 h in the order of nNOS> iNOS> eNOS as detected by immunostaining. Although Caspase-3 mRNA could be found in all groups, DNA fragmentation was only seen at 6 h and 24 h. The expressions of NOS and Caspase-3 mRNA were inhibited in the rat brain when dexamethasone was administrated.

CONCLUSIONLPS-induced NO production induces apoptosis of neurons through mechanism involving the Caspase-3 activation, which may play an important role in the pathogenesis of brain damage during endotoxemia, and neuro-protective effects of dexamethasone may be partially realized by inhibiting the expression of NOS mRNA.

Animals ; Animals, Newborn ; Apoptosis ; Brain ; drug effects ; enzymology ; Caspase 3 ; Caspases ; genetics ; metabolism ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Endotoxemia ; chemically induced ; enzymology ; Female ; Lipopolysaccharides ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; Nitric Oxide Synthase ; genetics ; metabolism ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type II ; Nitric Oxide Synthase Type III ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction

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

OBJECTIVETo investigate the feasibility of transfection of recombinant human endothelial nitric oxide synthase (eNOS) into human hypertrophic scar fibroblasts (HSFbs), and to observe NO secretion and the synthesis of collagen I and III.

METHODSRecombinant human eNOS with karyocyte expressive vector was constructed in vitro, then was transfected into HSFbs which was isolated from hypertrophic scar tissues and cultured in vitro (T group). The HSFbs untransfected (normal culture) or transfected with empty-vector was used as control group and empty-vector group respectively. The mRNA expression of eNOS, collagen I and III was determined by Realtime PCR. The content of NO was determined by NO assay kit.

RESULTSThe expression of eNOS mRNA in T group was 5.92 +/- 0.21, which was obviously higher than that in empty-vector group (0.98 +/- 0.13, P < 0.05). The expression of collagen I mRNA (0.76 +/- 0.15), and collagen III (0.79 +/- 0.08) in T group was significantly lower than those in empty-vector group (0.98 +/- 0.15, 1.02 +/- 0.12, P < 0.05, respectively). The content of NO in T group (36.1 +/- 0.8 micromol/L) was obviously higher than that in empty-vector group (28.4 +/- 1.0 micromol/L, P < 0.01) and control group (27.7 +/- 1.3 micromol/L, P < 0.01).

CONCLUSIONHSFbs can be the target cells for eNOS gene transfection. The transfected cells can express eNOS and produce NO, which inhibit the synthesis of collagen.

Cicatrix, Hypertrophic ; metabolism ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; metabolism ; Fibroblasts ; metabolism ; Humans ; In Vitro Techniques ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; genetics ; RNA, Messenger ; metabolism ; Transfection

AIMTo investigate the effect of icariin on erectile function and the expression of nitric oxide synthase (NOS) isoforms in castrated rats.

METHODSThirty-two adult male Wistar rats were randomly divided into one sham-operated group (A) and three castrated groups (B, C and D). One week after surgery, rats were treated with normal saline (groups A and B) or oral icariin (1 mg/[kg.day] for group C and 5 mg/[kg.day] for group D) for 4 weeks. One week after treatment, the erectile function of the rats was assessed by measuring intracavernosal pressure (ICP) during electrostimulation of the cavernosal nerve. The serum testosterone (ST) levels, the percent of smooth muscle (PSM) in trabecular tissue, and the expression of mRNA and proteins of neuronal nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS) and phosphodiesterase V (PDE5) in corpus cavernosum (CC) were also evaluated.

RESULTSICP, PSM, ST and the expression of nNOS, iNOS, eNOS and PDE5 were significantly decreased in group B compared with those in group A (P 0.01). However, ICP, PSM and the expression of nNOS and iNOS were increased in groups C and D compared with those in group B (P 0.05). Changes in ST and the expression of eNOS and PDE5 were not significant (P 0.05) in groups C and D compared with those in group B.

CONCLUSIONOral treatment with icariin ( 98.6 % purity) for 4 weeks potentially improves erectile function. This effect is correlated with an increase in PSM and the expression of certain NOS in the CC of castrated rats. These results suggest that icariin may have a therapeutic effect on erectile dysfunction.

3',5'-Cyclic-GMP Phosphodiesterases ; genetics ; metabolism ; Animals ; Blood Pressure ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; Drugs, Chinese Herbal ; pharmacology ; Erectile Dysfunction ; drug therapy ; metabolism ; Flavonoids ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Male ; Muscle, Smooth ; drug effects ; physiology ; Nitric Oxide Synthase ; genetics ; metabolism ; Nitric Oxide Synthase Type I ; genetics ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Orchiectomy ; Penile Erection ; drug effects ; Penis ; drug effects ; enzymology ; Pressure ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Testosterone ; blood

AIMTo investigate the effect of aging on the expression of nitric oxide synthase I (NOS I) and the activity of NOS in rat penis.

METHODSSixty male rats from 3 age groups (adult, old and senescent) were investigated. The expression of NOS I protein and mRNA in rat penis were detected by Western blot and RT-PCR respectively and the NOS activity, with ultraviolet spectrophotometry.

RESULTSIn the old and senescent group, NOS I protein expression was significantly decreased as compared with the adult. NOS I mRNA expression was well correlated with the protein expression. NOS activity was not statistically different between the adult and old groups, but it was significantly reduced in the senescent compared with the adult group (P<0.01).

CONCLUSIONThe aging-induced decreases in NOS I expression and NOS activity may be one of the main mechanisms leading to erectile dysfunction in the senescent rats.

Aging ; metabolism ; Animals ; Blotting, Western ; Male ; Nitric Oxide Synthase ; genetics ; metabolism ; Nitric Oxide Synthase Type I ; Penis ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

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