OBJECTIVES: The present study was designed to investigate the effect of ginseng saponin and its major active metabolite on the HPA axis under acute stress-i.c.v. injection stress, and immobilization stress, and to examine whether nitric oxide is involved in the mechanism of ginseng saponin on the HPA axis under acute stress. METHODS: In the experiment to study the effect of ginseng on HPA axis during stress, various dose of GTS were injected intracerebroventricularly(i.c.v.) or intraperitoneally(i.p.). Plasma corticosterone levels were measured 30 min after the i.c.v. injection stress. Immobilization stress was applied for 30 min and then blood was cellected for the assays of plasma corticosterone levels immediately after the completion of immobilization stress. To determine the active ginsenosides that can affect the stressinduced plasma corticosterone levels, various dose of each gisendosides(Rb1, Rb2, Rc, Re, Rf, Rg1, 20(S)-Rg3, and 20(R)-Rg3) were injected i.c.v. or i.p.. In the experiment to determine the involvement of the nitric oxide in the inhibitory effect of ginseng on the HPA, NG-Nitro-L-arginine methyl ester(L-NAME) and ginsenosides were coadministered i.c.v. or i.p., and plasma corticosterone levels were measured 30 min after stress was applied. RESULTS: First, the present study showed that ginseng total saponin, ginsenoside Rg3(S form), and ginsenoside Rc administered i.c.v. attenuated the intracerebroventricular injection stress-induced increase in plasma corticosterone levels, and these effects were removed by nitric oxide co-injection. Second, ginseng total saponin and ginsenoside Rc administered i.p. attenuated the immobilization stress-induced increase in plasma corticosterone levels, but ginsenoside Rg3(S form) did not attenuate the immobilization stress-induced increase in plasma corticosterone levels. The attenuative effects of ginseng total saponin and ginsenoside Rc in the immobilization stress-induced increase in plasma corticosterone levels were not affected by L-NAME co-injection. CONCLUSION: This study suggests that ginseng saponin attenuated stress-induced increase in plasma corticosterone levels and these effects were mediated by different mechanisms according to the components of ginseng saponin, and routes of administration.
Animals ; Axis, Cervical Vertebra ; Corticosterone* ; Ginsenosides ; Immobilization ; Mice* ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Nitroarginine ; Panax* ; Plasma* ; Saponins*

BACKGROUND: Etomidate produces minimal cardiovascular effects in clinical uses but their effects on the porcine coronary arteries are not well known yet. We studied the direct effects of etomidate on porcine coronary arterial tone and the underlying mechanism of its vascular relaxation. METHODS: Porcine coronary arterial ring segments (3-4 mm) with or without endothelium were suspended in modified Krebs solution (37oC) and preconstricted with K+ 40 mM. Changes in tension were measured following cumulative administrations of etomidate (10(-5), 5 x 10(-5) and 10(-4) M). Relaxation caused by etomidate (10(-4)M) were measured in the presence of either NG-nitro-L-arginine methyl ether (L-NAME 10(-5)M, n = 13), indomethacin (2.8 x 10(-5)M, n = 12), methylene blue (2 x 10(-5)M, n = 12), tetraethylammonium (TEA, KCa2+ blocker, 20 mM, n = 11), glibenclamide (n = 13) 2.5 x 10(-5)M or 4-aminopyridine (4-AP, K+ DR blocker, 10 4 M, n = 12). Effects of etomidate on Ca2+ influx through the voltage operated channel (VOC) of the vascular cells were also evaluated. RESULTS: Arterial reings were relaxed by etomidate in a concentration-dependent manner and these effects were not affected by endothelium. In the etomidate pretreated group, arterial ring in a calcium-free solution showed no contraction with KCl 40 mM, but the contraction after the administration of calcium 2.5 mM less than without etomidate group (47.2 +/- 8.7% vs 97.7 +/- 10.6%). The other group, not pretreated with etomidate, showed the same vascular tone of the control group in a slow upstroke manner with calcium administration. Pretreatment with either L-NAME, indomethacin and methylene blue did not affect etomidate-induced vasorelaxation. The TEA, glibenclamide and 4-AP pretreated groups also did not affect the vascular relaxation. CONCLUSIONS: Etomidate relaxes the porcine coronary artery in a concentration-dependent manner withor without endothelium, via inhibition of Ca2+ influx through the voltage-operated Ca2+ channel.
4-Aminopyridine ; Calcium ; Coronary Vessels* ; Endothelium ; Ether ; Etomidate* ; Glyburide ; Indomethacin ; Methylene Blue ; NG-Nitroarginine Methyl Ester ; Nitroarginine ; Relaxation ; Tea ; Tetraethylammonium ; Vasodilation

PURPOSE: To investigate the role of nitric oxide(NO) on the survival and proliferation of human Tenon capsule fibroblasts(HTCF) in tissue culture. METHODS: Following exposure to NO donor such as sodium nitroprusside(SNP) or to NO synthase inhibitor such as N omega-Nitro-L-arginine methyl ester(L-NAME) at various concentrations for 24 hr in the media with or without serum, the cellular proliferation and nitrite production were assessed by rapid colorimetric assay (MTT assay) and Griess assay, respectively. RESULTS: L-NAME decreased the cellular proliferation of HTCF in a dose-dependent manner. In contrast, SNP increased the proliferation in a dose-dependent manner. The differences of nitrite production were more remarkable in the serum-deprived condition with the L-NAME or SNP administration. CONCLUSIONS: NO donor has a proproliferative, and NOS inhibitor has a antiproliferative activities on the proliferation of HTCF. This suggests that NO may be act as an important modulator in wound healing.
Cell Proliferation ; Fibroblasts* ; Humans* ; NG-Nitroarginine Methyl Ester ; Nitric Oxide Synthase ; Nitric Oxide* ; Nitroarginine ; Nitroprusside ; Sodium ; Tenon Capsule* ; Tissue Donors ; Wound Healing

AIMS OF STUDY: During reflex micturition, the urethral outlet remains open (relaxed) to promote urinary emptying. The mechanisms involved in the relaxation of urethral outlet is thought to be complex including nitric oxide (NO) pathway and beta-adrenergic activity. The aims of the study focused on these several issues related to the neural control of urethral outlet in vivo. MATERIALS & METHODS: Female rats weighing 200~300 gm were anesthetized wish urethane. Catheters were inserted into femoral artery for drug administration.4 two-way catheter (16 G angiocath) was inserted into the bladder for saline infusion and pressure monitoring. A separate cannula (PE 50) was placed into the urethra via external urethral meatus or proximal urethrat opening to record urethral pressure. The bladder was filled with saline at a rate of 0.1 ml/min to induce reflex micturition. Urethral pressure was recorded via cannula through which saline was infused at a rate of 0.05 ml/min. Isovolumetric bladder contraction and urethral pressure were recorded simultaneously. After an equilibration period of 30 minutes, baseline intravesical and urethral pressure were recorded for 10 minutes prior to drug administration. NG-nitro-L-arginine methylester (L-NAME, 10 to 15 mg/kilrogram, i.v.), L-arginine (150 mg/kilrogram, i.v.), propranolol (1 microM., 0.1 ml/250 mg, i.a.), and phenylephrine (1 0~100 microM, i.a.) were administrated. RESULTS: During isovolumetric bladder contraction, urethral pressure was decreased simultaneously, and then returned to the resting states in conjunction with end of the bladder contraction. After the administration of L-NAME, the magnitude of reflex urethral relaxation was decreased significantly (42.6 +/- 15.1% of the control, p<0.01), and this effect was reversed by addition of L-arginine. Administration of propranolol also inhibited urethral relaxation (66.4% of the control). Administration of L-NAME followed by propranolol almost completely abolished the urethral relaxation. Administration of phenylephrine increased the resting urethral tone (mean; 4 cmH2O) significantly, and the magnitude of urethral relaxation was decreased substantially. CONCLUSION: These RESULTS suggest that urethral relaxation is mediated by several neural factors. NO seems like to a potent mediator in a reflex relaxation of the urethral smooth muscle during micturition. Also, beta-adrenergic stimulation play an important role for urethral relaxation. alpha-adrenergic nerve discharge, contributed to contraction of urethral smooth muscle, shows inhibitory effect against the reflex urethral relaxation.
Animals ; Arginine ; Catheters ; Female ; Femoral Artery ; Humans ; Muscle, Smooth ; NG-Nitroarginine Methyl Ester ; Nitric Oxide* ; Nitroarginine ; Phenylephrine ; Propranolol ; Rats ; Reflex ; Relaxation* ; Urethane ; Urethra ; Urinary Bladder ; Urination

The aim of this study was to evaluate neurotoxicity of Nitric oxide(NO) on cornea after excimer laser photorefractive keratectomy(PRK). PRK was performed on rabbit eyes. According to the time table, tear samples were collected with microcapillary tubes and corneal sensitivity was measured with a Cochet-Bonnet esthesiometer. No generation in the tear fluid was analyzed. To demonstrate NO Synthase(NOS), immunohistochemical localization was performed on frozen sections from rat eyeball tissue. Western blot analysis was used for detection of peroxynitrite, powerful oxidant of NO. NO generation was increased and reached to a maximum value(0.69+/-0.22micrometer/microgram) after 96 hours of PRK, as compared with in normal subjects(Mean: 0.30+/-0.08micrometer/microgram) and was not increased in the treated group with topical application of Ng-nitro-L-arginine methyl ester(L-NAME), a competitive inhibitor of constitutive NOS(cNOS) and inducible NOS(iNOS). Corneal sensitivity decreased below pretreatment levers after three postoperative days, but it was not observed in the L-NAME applied group. We have confirmed that a very strong iNOS and BNOS immunoreactivity was present in corneal keratocytes. Western blot analysis identifed the bands of nitrotyro-sine-proteins suggesting in vivo peroxynitrite toxicity. Our results suggested that NO generated from the enzyme after PRK decreased corneal sensitivity by damaging corneal sensory nerve through the NO and iths oxidant peroxitrite. Therefore topical application of a NOS inhibitor may be effective in maintaining corneal sensitivity.
Animals ; Blotting, Western ; Cornea ; Corneal Keratocytes ; Frozen Sections ; Lasers, Excimer* ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Nitroarginine ; Peroxynitrous Acid ; Photorefractive Keratectomy* ; Rats

BACKGROUND: It is generally accepted that propofol does not inhibit hypoxic pulmonary vasoconstriction (HPV). However, because the previous studies for the effects of propofol on HPV were established in vivo, the effects of physiologic variables could not be ruled out. Therefore, we investigated the effects of various concentrations of propofol on HPV at isolated rat lungs and the relationship of these effects of propofol on HPV and endothelium-derived relaxing factor (EDRF) and an ATP-dependent K+ channel which were candidates as the mechanism of HPV. METHODS: In 30 isolated rat lungs, after three hypoxic challenges for 5 minutes, we administered saline in the control group, N(G)-nitro-L-arginine methyl ester (L-NAME) in the L group and glibenclamide in the G group followed by three hypoxic challenges for 5 minutes. In addition, we studied the effects of various concentrations of propofol on HPV in the three groups. RESULTS: L-NAME and glibenclamide did not alter baseline pulmonary arterial pressure but L-NAME significantly enhanced HPV. Clinical concentrations of propofol did not affect HPV and high concentrations of propofol inhibited HPV. The pretreatment of L-NAME and glibenclamide did not alter the inhibition of HPV even at high concentrations of propofol. CONCLUSIONS: The EDRF and ATP-dependent K+ channel did not largely contribute to baseline pulmonary arterial tone but EDRF might be released and downregulate HPV. Clinical concentrations of propofol did not inhibit HPV but high concentrations of propofol inhibited HPV. In addition, the mechanism of inhibition of HPV at high concentrations of propofol did not relate to the EDRF pathway and ATP-dependent K+ channel.
Animals ; Arterial Pressure ; Endothelium-Dependent Relaxing Factors ; Glyburide* ; Lung* ; NG-Nitroarginine Methyl Ester* ; Propofol* ; Rats* ; Vasoconstriction*

The present study was aimed to investigate whether and to what extent hypertension affects the relaxation of the corpus cavernosum. The corpus cavernosum was isolated from 12-week 2- kidney, 1-clip hypertensive rats. The corporal strips were isolated and suspended longitudinally in an organ bath. They were precontracted with phenylephrine, and their responses to electrical field stimulation (EFS) were examined. EFS caused a frequency-dependent contraction (60%) or relaxation (40%) of the corpus cavernosum precontracted with phenylephrine. The contraction response was inhibited or abolished and only frequency-dependent relaxation appeared in the presence of atropine (0.00001mol/L) and guanethidine (0.00001mol/L). The relaxation response to EFS of the corporal preparation precontracted with phenylephrine was attenuated or abolished in the presence of L-NAME (0.0001mol/L). The corporal preparation from the hypertensive rats also showed a frequency-dependent relaxation, however, the degree of which was lower at a low frequency of stimulation than that from the normotensive control. These results suggest that endothelium-derived nitric oxide released upon neural stimulation partly mediate the relaxation of the corpus cavernosum. It is also suggested that hypertension is associated with a partly attenuated relaxation response to EFS.
Animals ; Atropine ; Baths ; Guanethidine ; Hypertension ; Kidney ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Phenylephrine ; Rats* ; Relaxation

PURPOSE: We investigated the effects of the superoxide radical on rat whole bladder contractility with duroquinone (superoxide radical generator, Dq) and diethyldithiocarbamate (superoxide dismutase inhibitor, DETCA), and the effects of ginseng saponin (GS) against superoxide radical injury. MATERIALS AND METHODS: Isometric tension changes of isolated rat whole bladders were recorded in an organ bath using a force transducer. The acute effects of Dq and Dq preincubated with DETCA were assessed on resting tension, electrical field stimulation, and bethanechol-, ATP-, and KCl-induced contraction. The effects of Dq and Dq preincubated with DETCA in the presence of sodium nitroprusside and GS were investigated. RESULTS: The resting tension of the muscle was not changed by Dq and Dq preincubated with DETCA. Dq had a harmful effect on only ATP- and KCl-induced detrusor contraction, whereas Dq pretreated with DETCA attenuated the induction of detrusor contraction which was reduced in response to the exogenous NO including GS. In the presence of L-NAME, the effects of GS reduced the Dq-induced inhibition on the detrusor contractility. CONCLUSIONS: It is suggested that the superoxide radical may be the cause of voiding difficulty. GS, as a NO synthesis stimulator, seems to act as a scavenger of the superoxide anion. However further study on the effect of each subfraction of GS is needed for clinical application.
Animals ; Baths ; Ditiocarb ; NG-Nitroarginine Methyl Ester ; Nitroprusside ; Panax* ; Rats* ; Saponins* ; Superoxides* ; Transducers ; Urinary Bladder*

PURPOSE: To investigate the effect of hypoxia on the survival and nitric oxide (NO) production of cultured trabecular meshwork (TM) cells. METHODS: After inducing chemical hypoxia with sodium cyanide, the survival and nitrite production of the primarily cultured porcine TM cells were assessed with MTT and Griess assays. The effect of NOS inhibitor, N(omega)-Nitro-L-arginine methyl ester (L-NAME), was also assessed. Flow cytometry using annexin/PI was done to evaluate apoptosis. RESULTS: Chemical hypoxia decreased TM cell survival significantly (p<0.05) with increased NO production. This hypoxia-induced antiproliferative effect was abolished by L-NAME (p<0.05). Flow cytometric analysis revealed that hypoxia induced apoptosis of TM cells, which was inhibited by L-NAME. CONCLUSIONS: Hypoxia decreases the survival of TM cells and induced apoptosis, accompanied by increased NO production. The hypoxia-induced decreased survival of TM cells may be mediated by NO.
Anoxia* ; Apoptosis ; Cell Survival ; Flow Cytometry ; NG-Nitroarginine Methyl Ester ; Nitric Oxide* ; Sodium Cyanide ; Trabecular Meshwork*

OBJECTIVE: Early maternal separation (EMS) during the development has been known to influence the alteration of behavior in adulthood. Nitric oxide (NO) may have been implicated to play a crucial role in the neurodevelopment as an intracellular and intercellular messenger. This study was designed to investigate the neurochemical mechanism of the behavioral changes resulting from EMS during the development in juvenile rats. METHODS: Experimental group consisted of subjects that were removed and weaned from the day on postnatal day 15. Control group were the litters that experienced no EMS until postnatal day 21. On postnatal day 15 and 36, the locomotor activity (LA) was measured. On postnatal day 36 the behavioral changes in the forced swimming test (FST) were also measured. Test drugs were intraperitoneally injected including MK-801 (0.5 mg/kg), N omega-nitro-L-arginine (L-NA, 20 mg/kg), paroxetine (20 mg/kg), and bupropion (150 mg/kg). RESULTS:EMS produced the decrease of LA significantly in juvenile rats (p<0.001). Both MK-801 and L-NA increased LA in experimental group (p<0.001) and control group (p<0.05). The degree of increase was higher in experimental group than in control group. However, both paroxetine and bupropion increased LA in experimental group (p<0.001, p<0.05), but not in control group. In the FST, immobility was significantly increased in experimental group compared with control group (p<0.001). The increases of immobility in experimental group were abolished after injecting MK-801, L-NA, paroxetine, and bupropion, respectively. CONCLUSION: These results indicate that EMS during the development can lead to behavioral abnormalities in juvenile rats. The underlying neurochemical mechanism of this behavioral changes may be, in part, related to the glutamatergic NMDA-NO pathway. This suggests that glutamatergic NMDA-NO pathway vulnerable to stress may predispose to depression.
Animals ; Bupropion ; Depression ; Dizocilpine Maleate ; Motor Activity ; Nitric Oxide ; Nitroarginine ; Paroxetine ; Rats ; Swimming