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Objective To investigate the biological effects of the Mucuna pruriens (M. pruriens) seed extracts that lacked L-DOPA, which was formerly reported as the active ingredient, on erectile dysfunction (ED) both in vitro and in vivo. Methods Seed of M. pruriens plant that cultivated in Mae Taeng District, Chiang Mai Province, Thailand, was collected. Component of its seeds were extracted and isolated into 2 fractions using methanol, polar and nonpolar. Each fraction was investigated for phytochemicals using gas chromatography and mass spectroscopy and was screened for biological activity in vitro using three different cell lines. The most biological active fraction was used to treat both streptozotocin (STZ)-induced diabetes mellitus-erectile dysfunction (DM-ED) male Wistar rats and normal rats (n = 6 per groups) to compare the effect on sexual behavior parameters, including number of intromission, mounting and ejaculation, with that of rats given Sildenafil by individually pairing with their female counterparts. Penile tissues and serums were collected to determine histological structure, related gene expression and biomolecules. Results The phytochemicals of the polar fraction were possibly catechol and its derivatives plus polyphenols, whereas the nonpolar fraction consisted of lipid derivatives. L-DOPA was not detected in either of the extracts. The polar fraction was able to up-regulate the expression of ED-related genes including eNOS and nNOS in vitro which subsequently promotes nitric oxide production and maintains intracellular cyclic guanosine monophosphate levels. When administrated to DM-ED rats, the polar extract significantly improved all sexual behavior parameters in DM-ED rats compared to untreated group (18.3 ± 1.8 to 10.8 ± 2.9 for intromission, 9.8 ± 2.2 to 5.7 ± 1.3 for mounting, and 1.8 ± 0.6 to 0.2 ± 0.4 for ejaculation). That effect might due to the ability of the extract to stimulate the expression of eNOS and nNOS which results in nitric oxide production and subsequently maintains cyclic guanosine monophosphate levels in penile tissue. Moreover, this extract may also prevent penile tissue deterioration due to diabetes. Conclusions The polar extract of M. pruriens seed can be used for ED therapy, especially in patients with metabolic diseases including diabetes. The action of the extract might be due to catechol and its derivatives and polyphenols.
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OBJECTIVE@#To investigate the biological effects of the Mucuna pruriens (M. pruriens) seed extracts that lacked l-DOPA, which was formerly reported as the active ingredient, on erectile dysfunction (ED) both in vitro and in vivo.@*METHODS@#Seed of M. pruriens plant that cultivated in Mae Taeng District, Chiang Mai Province, Thailand, was collected. Component of its seeds were extracted and isolated into 2 fractions using methanol, polar and nonpolar. Each fraction was investigated for phytochemicals using gas chromatography and mass spectroscopy and was screened for biological activity in vitro using three different cell lines. The most biological active fraction was used to treat both streptozotocin (STZ)-induced diabetes mellitus-erectile dysfunction (DM-ED) male Wistar rats and normal rats (n = 6 per groups) to compare the effect on sexual behavior parameters, including number of intromission, mounting and ejaculation, with that of rats given Sildenafil by individually pairing with their female counterparts. Penile tissues and serums were collected to determine histological structure, related gene expression and biomolecules.@*RESULTS@#The phytochemicals of the polar fraction were possibly catechol and its derivatives plus polyphenols, whereas the nonpolar fraction consisted of lipid derivatives. l-DOPA was not detected in either of the extracts. The polar fraction was able to up-regulate the expression of ED-related genes including eNOS and nNOS in vitro which subsequently promotes nitric oxide production and maintains intracellular cyclic guanosine monophosphate levels. When administrated to DM-ED rats, the polar extract significantly improved all sexual behavior parameters in DM-ED rats compared to untreated group (18.3 ± 1.8 to 10.8 ± 2.9 for intromission, 9.8 ± 2.2 to 5.7 ± 1.3 for mounting, and 1.8 ± 0.6 to 0.2 ± 0.4 for ejaculation). That effect might due to the ability of the extract to stimulate the expression of eNOS and nNOS which results in nitric oxide production and subsequently maintains cyclic guanosine monophosphate levels in penile tissue. Moreover, this extract may also prevent penile tissue deterioration due to diabetes.@*CONCLUSIONS@#The polar extract of M. pruriens seed can be used for ED therapy, especially in patients with metabolic diseases including diabetes. The action of the extract might be due to catechol and its derivatives and polyphenols.
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AIM To investigate the effects of Shuxuening Injection (Ginkgo biloba leaf extract) on serum lactic acid (Lac),soluble CD14-st (Presepsin) and nitric oxide synthase (NOS) levels in sepsis patients.METHODS One hundred and eight patients with sepsis treated by routine treatment in our hospital from Jan.2014 to Oct.2016 were randomly divided into two groups,control group and Shuxuening group (therapy group).Two weeks were one therapeutic course.Before the treatment (the onset of patients within 3 hours),at 6 hours and 5 days after the treatment,Lac and Presepsin levels were detected,and the changes of nitric oxide (NO),NOS,inducible nitric oxide synthase (iNOS) and sequential organ failure assessment (SOFA) score were observed.Incidence of major adverse cardiac events (MACE) and 28-day survival were recorded at the same time.RESULTS Before the treatment,there were no significant differences in SOFA score and the levels of Lac,Presepsin,NO,NOS and iNOS between the two groups (P ＞ 0.05).Six hours after the treatment,the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05),both the two groups had lower levels of Lac and Presepsin than those before the treatment (P ＜ 0.05);five days after the treatment,the levels of Lac and Presepsin in the two groups were lower than those at 6 hours after the treatment (P ＜ 0.05),the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05).The SOFA score,NO,NOS and iNOS levels after the treatment in the therapy group were lower than those in the control group (P ＜ 0.05).The levels of Lac and Presepsin in sepsis patients were positively correlated with SOFA score (r =0.245,0.261,P =0.011,0.006).The patients in the therapy group had lower incidence of MACE and 28-day mortality rate than those in the control group (P ＜ 0.05).CONCLUSION The therapeutic effect of Shuxuening Injection combined with routine treatment on sepsis patients is superior to that of routine treatment,which can improve the prognosis of patients to a certain extent.
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AIM To investigate the effects of Shuxuening Injection (Ginkgo biloba leaf extract) on serum lactic acid (Lac),soluble CD14-st (Presepsin) and nitric oxide synthase (NOS) levels in sepsis patients.METHODS One hundred and eight patients with sepsis treated by routine treatment in our hospital from Jan.2014 to Oct.2016 were randomly divided into two groups,control group and Shuxuening group (therapy group).Two weeks were one therapeutic course.Before the treatment (the onset of patients within 3 hours),at 6 hours and 5 days after the treatment,Lac and Presepsin levels were detected,and the changes of nitric oxide (NO),NOS,inducible nitric oxide synthase (iNOS) and sequential organ failure assessment (SOFA) score were observed.Incidence of major adverse cardiac events (MACE) and 28-day survival were recorded at the same time.RESULTS Before the treatment,there were no significant differences in SOFA score and the levels of Lac,Presepsin,NO,NOS and iNOS between the two groups (P ＞ 0.05).Six hours after the treatment,the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05),both the two groups had lower levels of Lac and Presepsin than those before the treatment (P ＜ 0.05);five days after the treatment,the levels of Lac and Presepsin in the two groups were lower than those at 6 hours after the treatment (P ＜ 0.05),the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P ＜ 0.05).The SOFA score,NO,NOS and iNOS levels after the treatment in the therapy group were lower than those in the control group (P ＜ 0.05).The levels of Lac and Presepsin in sepsis patients were positively correlated with SOFA score (r =0.245,0.261,P =0.011,0.006).The patients in the therapy group had lower incidence of MACE and 28-day mortality rate than those in the control group (P ＜ 0.05).CONCLUSION The therapeutic effect of Shuxuening Injection combined with routine treatment on sepsis patients is superior to that of routine treatment,which can improve the prognosis of patients to a certain extent.
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Objective To compare the expression of nitric oxide synthase (NOS ) in patients with gastroesophageal reflex disease (GERD)and healthy controls.Methods The distribution and relative protein amount of two NOS subtypes (nNOS and iNOS)were determined with immunohistological method,and their mRNA levels were measured with real-time PCR method.Results The nNOS and iNOS were mostly distributed in the cytoplasm in epithelia of esophageal mucosa.The nNOS and iNOS in reflux esophagitis (RE)were significantly higher than in non-erosive reflux disease (NERD)patients and healthy controls (P <0.05 ).The mRNA levels of nNOS and iNOS were also significantly higher in RE patients than in NERD patients and healthy controls (P <0.05).Conclusion The expressions of nNOS and iNOS were increased in the esophagus of RE patients,which may be related to the effects of NO on the onset of GERD.
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The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 (3~30 microM), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 (10 microM) also time-dependently inhibited the CA secretion evoked by DMPP (100 microM, a selective neuronal nicotinic receptor agonist) and high K+ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 (50 microg/mL), the secretory responses of CA evoked by veratridine (a selective Na+ channel activator (50 microM), Bay-K-8644 (an L-type dihydropyridine Ca2+ channel activator, 10 microM), and cyclopiazonic acid (a cytoplasmic Ca2+-ATPase inhibitor, 10 microM) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 (10 microM) and L-NAME (an inhibitor of NO synthase, 30 microM), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 (10 microM) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of Ca2+ and Na+ into the adrenomedullary chromaffin cells and also by suppressing the release of Ca2+ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.
Subject(s)
Animals , Rats , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester , Adrenal Medulla , Calcium , Catecholamines , Chromaffin Cells , Cytoplasm , Dimethylphenylpiperazinium Iodide , Membranes , Neurons , NG-Nitroarginine Methyl Ester , Nitric Oxide Synthase , Receptors, Nicotinic , Veins , VeratridineABSTRACT
@#Objective To study the effect of Tanshinone ⅡA(Tan ⅡA) on the contents of nitrous oxide(NO) and the activities of nitric oxide synthase(NOS) and immunologic NOS(iNOS) following cerebral ischemia reperfusion(I/R)injury in rats.Methods 40 Wistar rats were randomly divided into 4 groups,which were sham group,I/R group,low dosage Tan ⅡA treated group and high dosage Tan ⅡA treated group.The focal middle cerebral artery occlusion(MCAO) model was made.Rats were pretreated with Tan ⅡA for 3 d respectively before MCAO.After 90 min MCAO following 24 h of reperfusion,HE staining was investigated.The contents of NO and the activities of NOS and iNOS were also investigated.Results The change of ischemic impairment in low or high dosage Tan ⅡA treated group was lighter than that of I/R group,and high dosage Tan ⅡA treated group was lighter than that of low dosage Tan ⅡA treated group.Compared with the sham group,the contents of NO and the activities of NOS and iNOS increased in the ischemic territory(P<0.05).Compared with I/R group,low and high dose Tan ⅡA treated group reduced the contents of NO and the activities of NOS and iNOS dose-dependently(P<0.05).Conclusion Tan ⅡA may reduced cerebral ischemia-reperfusion injure by reducing the contents of NO and the activities of NOS and iNOS dose-dependently.
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OBJECTIVE: Resveratrol, a polyphenolic phytoalexin, is extracted abundantly from the red wine and grapes and biosynthesized as a defense agent to infection, ultraviolet and ozon etc. Recently, The cancer-preventive, anti-inflammatory and anti-oxidative effects of resveratrol have been reported. The aim of this study was to investigate the effect of resveratrol on the expression of nitric oxide synthases in hypoxic-ischemic brain injury in the neonatal rat model. METHODS: Embryonic cortical neuronal cell culture of rat brain was performed with pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for in vitro approaches. In addition, unilateral carotid artery ligation was induced in seven-days old neonatal rats for in vivo approaches. The real-time PCR using iNOS, eNOS and nNOS primer, and the western blotting using the same antibodies were done to identify the effects of resveratrol. RESULTS: The expression of iNOS, eNOS and nNOS in both cell culture and animal model of neonatal HI brain injury revealed that, as indicated by western blotting and real-time PCR, the expression of iNOS was decreased in the hypoxia group while those of eNOS and nNOS were increased in the hypoxia group compared with the normoxia group. The expression of iNOS was increased in the resveratrol-treated group while those of eNOS and nNOS decreased in the resveratrol-treated group compared with a hypoxic group. CONCLUSION: The present study demonstrates resveratrol might affect nitric oxide synthases expression in HI injury of the perinatal period
Subject(s)
Animals , Pregnancy , Rats , Hypoxia , Antibodies , Blotting, Western , Brain , Brain Injuries , Carotid Arteries , Cell Culture Techniques , Ligation , Models, Animal , Neurons , Nitric Oxide , Real-Time Polymerase Chain Reaction , Sesquiterpenes , Stilbenes , Vitis , WineABSTRACT
@#Objective To investigate the neuronal nitric oxide synthase (nNOS) distribution in intramural striated sphincter of female rats. Methods 4 female Wistar rats were anesthetized with chloral hydrate. The whole bladder and urethra specimen were harvested, fixed in formalin and imbedded in paraffin wax. Longitudinal and transverse sections were developed with 2 urethra respectively. Routine HE staining and Mallory phosphotungstic acid-hematoxylin staining were used for observation of urethral microstructure. nNOS distribution in intramural striated sphincter was determined with immunohistochemical staining. Results The urethras of female Wistar rat were tubular structure about 1.5～2.0 cm long. From outside inward,the structure included circular striated muscle layer,circular smooth muscle layer,longitudinal smooth muscle layer,dense connective tissue and epithelium. The longitudinal sections showed intramural striated muscle predominates the middle and distal third,not the whole urethral length. The transverse sections showed intramural striated muscle is closed ring shape,about 5～10 layers uneven distributed with more layers in the posterior wall than in the front wall. Strong positive expression of nNOS immunoreactivity was found in some intramural striated muscle fiber. Conclusion nNOS expresses in some intramural striated muscle fiber, surggesting nitric oxide may play an important role on intramural striated sphincter.
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PURPOSE: Mycophenolic acid (MPA), a potent inhibitor of inosine-monophosphate dehydrogenase (IMPDH), was used as a new immunosuppressive drug since 1990s. It was reported that MPA increased neuronal survival after excitotoxic injury, induced apoptosis in microglial cells, inhibited the induction of inducible nitric oxide synthase (iNOS) in astrocytes. and inhibited microglial cell proliferation in N-methyl-D-aspartate (NMDA) induced hippocampal cells. However, the effects of MPA on the perinatal hypoxic-ischemic (HI) brain injury had not been yet evaluated. Therefore, we examined whether MPA could be neuroprotective in the HI brain injury. METHODS: Cortical cells were cultured using a 18-day-pregnant Sprague-Dawley (SD) rats and incubated in 1% O2 incubator for hypoxia. MPA (10 ug/mL) before or after a HI insult were treated. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2.5 hours of hypoxic exposure (8% O2). MPA (10 mg/kg) were administrated intraperitoneally before or after a HI insult. Nitric oxide (NO) activity and expression of N-methyl-D-aspartate (NMDA) receptors also measured using Real-time PCR with primer pairs of isoforms of NOS; iNOS, endothelial NOS (eNOS), neuronal NOS (nNOS), and subunits of NMDA receptors; NR1, NR2A, NR2B, NR2C, NR2D. RESULTS: The expression of iNOS was decreased in the hypoxia group but increased in the MPA-treated group. However express or that eNOS and nNOS were inversed. The expression of all NMDA receptor subunits except NR2B was decreased in the hypoxia group but increased in the MPA-treated group. CONCLUSION: This study indicates that the administration of MPA before a HI insult could significantly protect against perinatal HI brain injury via some parts of NO-mediated or excitotoxic mechanisms.
Subject(s)
Animals , Rats , Hypoxia , Apoptosis , Astrocytes , Brain Injuries , Brain , Cell Proliferation , Incubators , Mycophenolic Acid , N-Methylaspartate , Neurons , Nitric Oxide Synthase Type II , Nitric Oxide Synthase , Nitric Oxide , Oxidoreductases , Protein Isoforms , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptors, N-Methyl-D-AspartateABSTRACT
@#ObjectiveTo investigate the effects of nitric oxide synthase (NOS) genetic transfection on the intimal hyperplasia of venous autografts. MethodsThe external jugular veins were autografted into abdominal aorta arteries in 20 Wistar rats, which were divided evenly into experimental or control groups. The transplanted veins of experimental group were immersed in the adenovirus-mediated eNOS gene solution for 15 minutes just before anastomosis. The transplanted vascular samples were taken out 2 weeks after operation. The intimal thickness(IH), degree of restenosis(DR), expression of PCNA and NOS mRNA were determined with histology and transcription polymerase chain reaction (PCR). ResultsThe IH, DR and PCNA decreased, while the expression of eNOS mRNA increased comparing with control group(P<0.01). ConclusionTransfection of NOS gene can inhibit the intimal hyperplasia of venous autografts.
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@# ObjectiveTo observe the curative effects of nerve growth factor (NGF) on experimental acute cerebral thromboemblia rats and study the mechanisms preliminarily.Methods24 model rats were randomly divided into three groups treated respectively with NGF, citicoline sodium (CS) and normal saline (NS) for 20 days, and the neurological grades of animals were observed before and after treatment. Then, 55 rats were randomly divided into three groups: the treated group (25 model rats, treated with NGF), control group (25 model rats, treated with NS) and normal group (5 normal rats, without treatment), the levels of nitric oxide synthase (NOS) of all animals were measured at 1 h, 3 h, 6 h, 12 h and 24 h after acute cerebral thromboemblia established.ResultsThe neurological grades of both NGF and CS treated groups were significantly lowered after treatment compared with NS control group ( P<0.05). NOS levels of cerebral thromboemblia areas were higher than that in the control group 1 hour, 3 hours after acute cerebral thromboemblia, the levels of NOS in NGF treatment group were obviously lower than that in the control group post-traumatic 1 hour, 3 hours and 6 hours.ConclusionNGF can accelerate the nervous function recovery of the rat with acute cerebral thromboemblia, the mechanisms is that NGF prohibits neurotoxicity of NOS.
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Endogenous nitric oxide (NO) has been known to regulate the salivary secretion and glandular blood flow. However, nitric oxide synthase (NOS) responsible for NO synthesis has not been well studied in lacrimal glands. The present study was aimed to investigate the distribution of nitric oxide synthase isoforms (endothelial, neuronal, and inducible NOS). Immunohistochemistry, using monoclonal mouse anti-endothelial NOS, anti-neuronal NOS, and anti-inducible NOS, was performed in exorbital lacrimal glands of the rat. Endothelial NOS (eNOS)-positive immunoreactivity was observed in vascular endothelium, intralobular duct and interlobular duct of the exorbital lacrimal gland of the rats, and also in the 3 major salivary glands of the rat. eNOSpositive immunoreactivity was most prominent in the intralobular and interlobular duct was well concentrated in cytoplasm of columnar epithelial duct cell. However, eNOS-positive immunoreactivity of the intercalated duct and serous acinus was absent. Neuronal NOS (nNOS)-positive immunoreactivity was seen in ganglion cells of exorbital lacrimal gland. iNOS or nNOS-positive immunoreactivy was not detected either in excretory ducts or in acinar cells. Inducible NOS-positive immunoreactivity was not seen. There results reveal the presence of eNOS and nNOS in the exorbital lacrimal gland, which may be related with regulation of the glandular secretion and blood flow.
Subject(s)
Animals , Mice , Rats , Acinar Cells , Cytoplasm , Endothelium, Vascular , Ganglion Cysts , Immunohistochemistry , Lacrimal Apparatus , Neurons , Nitric Oxide Synthase , Nitric Oxide , Protein Isoforms , Salivary GlandsABSTRACT
Aim To study the role of nitric oxide by measuring the expression and activity of nitric oxide synthase in hippocampus of rats tolerance and dependence to the anticonvulsant action of flurazepam.Method Animal models of toleran and dependence to flurazepam were established. Nitric oxide synthase in hippocampus was measured using Western blot and immunohistochemistry, and its activity was evaluated using colorimetry. Result Hippocampal nNOS increased in rats tolerance to flurazepam. In rats dependence to flurazepam, hippocampal nNOS increased but without statistical significance. Total NOS activity increased dramatically both in flurazepam tolerane and dependene rats.Conclusion NO may play a role in rats tolerance and dependence to the anticonvulsant action of flurazepam.
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Objective To explore the effects of Ganoderma spore(germination activated Ganoderma spore) on survival and expression of neurotrophin_3(NT_3) and nitric oxide synthase(NOS) of injured motor neurons in rat spinal cord. Methods Different dosages of Gandoerma spore were irrigated into the rat's stomach respectively in experimental groups after oneside ventral root cut.The survival ratio of injured motor neurons was counted.NT_3 expression of surviving motor neurons was measured by immunohistochemistry and in situ hybridization methods and NOS activity was measured by enzymo_histochemistry method. Results Thirty_five days after ventral root cut,the survival ratio of motor neurons was 47.32% in control group,and those were 67.11%,72.67% and 81.67% respectively in low,medial and high dosage Ganoderma spore groups.Expressions of NT_3 and NOS of surviving motor neurons in high dosage Ganoderma spore group were higher than those in control group.Conclusion Ganoderma spore may promote the survival of injured spinal motor neurons,and survival ratio of injured motor neurons is related to the dosages of Ganoderma spore.Ganoderma spore may also enhance the expressions of NT_3 and NOS of surviving spinal motor neurons.
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The role of nitric oxide (NO) in the ocular surface remains unknown. We investigated the conditions leading to an increase of NO generation in tear and the main sources of NO in ocular surface tissue. We evaluated the dual action (cell survival or cell death) of NO depending on its amount. We measured the concentration of nitrite plus nitrate in the tears of ocular surface diseases and examined the main source of nitric oxide synthase (NOS). When cultured human corneal fibroblast were treated with NO producing donor with or without serum, the viabilities of cells was studied. We found that the main sources of NO in ocular surface tissue were corneal epithelium, fibroblast, endothelium, and inflammatory cells. Three forms of NOS (eNOS, bNOS, and iNOS) were expressed in experimentally induced inflammation. In the fibroblast culture system, the NO donor (SNAP, S-nitroso-N-acetyl-D, L-penicillamine) prevented the death of corneal fibroblast cells caused by serum deprivation in a dose dependent manner up to 500 micrometer SNAP, but a higher dose decreased cell viability. This study suggested that NO might act as a doubleedged sword in ocular surface diseases depending on the degree of inflammation related with NO concentration.
Subject(s)
Animals , Humans , Rabbits , Apoptosis/drug effects , Aqueous Humor/metabolism , Blood Proteins/pharmacology , Cell Survival/drug effects , Cells, Cultured , Epithelium, Corneal/cytology , Fibroblasts/cytology , Nitric Oxide/biosynthesis , Nitric Oxide Donors/pharmacology , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type I , Nitric Oxide Synthase Type II , Nitric Oxide Synthase Type III , Penicillamine/analogs & derivatives , Peroxynitrous Acid/biosynthesis , Tears/metabolism , Uveitis/metabolismABSTRACT
PURPOSE: NO, a diatomic free radical, plays a diverse physiological and pathophysiological roles in the vascular, neuronal and immune systems. It is produced by nitric oxide synthase (NOS) which consists of three different isoforms. In this study we investigated NOS expression in 84 human breast carcinomas and its associations to other clinicopathological factors. METHODS: The immunohistochemical staining for NOS expression in 84 human breast carcinomas were performed and their medical records were reviewed retrospectively. RESULTS: iNOS expression in tumor cells was observed in 48.2% and eNOS expression was detected in 51.9%. iNOS expression in tumor cells has positive correlation with eNOS expression in tumor and is associated with iNOS expression in stroma and endothelial cells. Although iNOS expression in tumor cells has negative correlation with tumor size (P=0.047) and lymph node metastasis (P=0.002), it has no effects on 5 year overall and disease free survivals. iNOS expression in stroma also has negative correlation with tumor size (P=0.016) and nuclear grade (P=0.025). No significant correlation between eNOS expression and clinicopathological factors was observed but eNOS expression in tumor cells contributed to worse 5 year overall survivals (92.1% vs 77.0%) in marginal significance (P=0.053). CONCLUSION: These data suggest that iNOS expression in tumor may have an inhibitory effect in tumor growth and lymph node metastasis. These results may be further investigated.
Subject(s)
Humans , Breast Neoplasms , Breast , Diatoms , Endothelial Cells , Immune System , Lymph Nodes , Medical Records , Neoplasm Metastasis , Neurons , Nitric Oxide Synthase , Nitric Oxide , Protein Isoforms , Retrospective StudiesABSTRACT
Nitric oxide (NO) is a diffusible cellular mediator generated by the enzyme nitric oxide synthase (NOS). NO involvement has been demonstrated in mechanisms of synaptic plasticity, particularly in hippocampal long-term potentiation, a mechanism that underlies certain forms of learning and memory. Several data have shown that NO production is decreased in the aged rat. Changes in the nNOS mRNA-containing neurons with aging were demonstrat-ed by in situ hybridization. nNOS mRNA-positive cells in aged rats were present in all cortical areas, hippocampus, and cerebellum and the distribution was similar to that of the young adult group. The number of nNOS mRNA-positive cells was significantly decreased in the occipital (86.2%), parietal (81.2%), and temporal cortices (79.4%), also in hippocampal CA1 (86.2%), dentate gyrus (92.3%), and cerebellar Purkinje cells (73.9%). The most severe decrease was found in hippocampal area. These results are consistent with the former studies showing NO decrease in aging brain and nNOS mRNA decrease indicates the involvement of neuronal system containing NOS in the aging brain, especially for learning and memory.
Subject(s)
Animals , Humans , Rats , Young Adult , Aging , Brain , Cerebellum , Cerebral Cortex , Dentate Gyrus , Hippocampus , In Situ Hybridization , Learning , Long-Term Potentiation , Memory , Neurons , Nitric Oxide , Nitric Oxide Synthase , Plastics , Purkinje Cells , Rabeprazole , RNA, MessengerABSTRACT
The role of nitric oxide (NO) in ocular surface diseases remains unknown. We investigated the conditions leading to increase NO generation in tears and the main sources of ocular surface tissue. We evaluated the possibility of a dual action (cell survival or cell death) depending on the amount of NO. The concentration of nitrite plus nitrate, the stable end-product of NO, was measured in the tears of various ocular surface diseases. We also examined the main source of nitric oxide synthase (NOS) using immunohistochemical staining & Western blot analysis. When cultured human corneal fibroblasts were treated with NO producing donor with or without serum, the viability of cells was studied. We found that sources of NO in ocular surface tissue primarily included corneal epithelium, fibroblasts, endothelium and inflammatory cells. Three forms of NOS (eNOS, bNOS, & iNOS) were expressed in experimentally induced inflammation. Cell death by NO revealed TUNEL positive staining, however in the EM finding, this NO specific cell death was an atypical necrosis showing perinuclear large vacuolization and mitochondrial swelling. In the fibroblasts culture system, the NO donor (SNAP, S-nitroso-N-acetyl-D, L-penicillamine) prevented the death of corneal fibroblasts caused by serum deprivation in a dose dependent manner up to 500 m SNAP, although a higher dose decreased cell viability. This study suggested that NO might act as a double-edged sword in ocular surface disease depending on the degree of inflammatory condition related with NO concentration.
Subject(s)
Humans , Animals , Cells, Cultured , Cornea/metabolism , Eye Diseases/physiopathology , Nitric Oxide/metabolism , Tears/metabolismABSTRACT
Nitric oxide (NO) involvement has been demonstrated in mechanisms of synaptic plasticity, particularly in hippocampal long-term potentiation, a mechanism that underlies certain forms of learning and memory. Further, NO has been shown to regulate various neurotransmitters which play an important role in learning and memory. Several findings suggest that NO production may be decreased in the aged rat. Changes in the nNOS-containing neurons with aging were demonstrated by immunocytochemistry and in situ hybridization. NOS-immunoreactive cells in aged rats were present in all cortical areas and the hippocampus, and the pattern of distribution was similar to that of the control group. The number of NOS-immunoreactive cells in the cerebral cortex was significantly decreased in the aged rats, but the extent of changes was variable in each area, and ranged from mild decrease (50%). Severely decreased areas were the cingulate cortex, parietal cortex area 1, temporal cortex area 1, 2, 3, medial part of occipital cortex area 2, monocular and binocular part of occipital cortex area 1, entorhinal cortex, hippocampus proper, dentate gyrus and subiculum. Moderately decreased areas (30~50%) were frontal cortex area 1, 2, 3, parietal cortex area 2, forelimb, hindlimb, lateral part of occipital cortex area 2. Slightly decreased area was insular cortex. Morphologically, the number of dendritic branches seemed to be decreased in aged group and the length of dendrites of NOS-IR neurons showed a tendency to shorten. These results indicate the involvement of neuronal system containing NOS in the aging brain, and provide the first morphological evidence for the loss of NOS neurons in the cerebral cortex of the aged rats by immunocytochemistry. Further multidisciplinary investigations involving normal aging and neurodegenerative disease such as Alzheimer's disease are needed to clarify the importance of nitric oxide changes in the cerebral cortex with aging.