• The Korean Journal of Pharmacology
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• v.31 no.2
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• pp.233-239
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• 1995

To verify the effect of immunosuppressants on the endotoxin-induced increase in iNOS activity, the action of immunosuppressants, dexamethasone (1.5 mg/kg), azathioprine (5 mg/kg/day) and cyclosporine (10 mg/kg), were evaluated in mice pretreated with LPS. The intraperitoneal injection of lipopolysaccharide (10 mg/kg) increased the nitric oxide synthase (NOS) activity in the brain and liver to maximum at 1 and 3 hours, respectively. The increase in NOS activity was blocked by the treatment with NOS inhibitor, LNAME(300 mg/kg) and aminoguanidine(100 mg/kg); a protein inhibitor, cycloheximide (10 mg/kg); and a transcription inhibitor of inducible NOS(iNOS), dexamethasone(1.5 mg/kg). Immunosuppressants, azathioprine (5 mg/kg) and cyclosporine (10 mg/kg), effectively blocked the increase in NOS activity. These results suggest that iNOS expression plays an important role in LPS-induced the increase in NOS activity and that immunosuppressants can be used as candidate for therapeutic agents in endotoxemia.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.6
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• pp.337-341
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• 2006

The present study was designed to investigate the protein expression of nitric oxide synthase (NOS) and guanylyl cyclase (GC) activity in ischemia/perfusion (I/R) renal injury in rats. Renal I/R injury was experimentally induced by clamping the both renal pedicle for 40 min in Sprague-Dawley male rats. The renal expression of NOS isoforms was determined by Western blot analysis, and the activity of guanylyl cyclase was determined by the amount of guanosine 3', 5'-cyclic monophosphate (cGMP) formed in response to sodium nitroprusside (SNP), NO donor. I/R injury resulted in renal failure associated with decreased urine osmolality. The expression of inducible NOS (iNOS) was increased in I/R injury rats compared with controls, while endothelial NOS (eNOS) and neuronal NOS (nNOS) expression was decreased. The urinary excretion of NO metabolites was decreased in I/R injury rats. The cGMP production provoked by SNP was decreased in the papilla, but not in glomerulus. These results indicate an altered regulation of NOS expression and guanylyl cyclase activity in I/R-induced nephropathy.

• Korean Journal of Acupuncture
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• v.30 no.4
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• pp.264-271
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• 2013

Objectives : To observe the changes in the expression of neurotransmitters NO and enzymes that create NO, such as nNOS, iNOS and eNOS, upon the needle insertion on river point, one of the five transport points of three yang meridians on the forefoot. Methods : Based on rats, needle was inserted on both sides of LI5, SI5 and TE6, which are river points of three yang meridians on the forefoot, and were retained for five minutes. After the retention, blood was drawn via cardiac puncture and tissues from each point around meridian vessels were extracted to be examined on the changes of the expression of NO, as well as of nNOS, iNOS and eNOS. Results : In terms of the effect on NO creation in tissues, none of the experimental groups showed any significant change compared to the Normal group. In terms of the effect on expression of nNOS within tissues, LI5 and SI5 showed significant increase based on the results of immunohistochemistry. In iNOS within tissues, LI5 and SI5 showed significant increase based on the results of immunohistochemistry. In eNOS within tissues, SI5 showed significant increase based on the results of immunohistochemistry. Conclusions : The effect on the function of NO, nNOS, iNOS and eNOS of needle insertion on the river points of three yang meridians on the forefoot could be observed, and based on this study, it is considered that the effect of needle stimulation on the changes of nervous system could be found out through additional research.

• Korean Journal of Acupuncture
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• v.30 no.1
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• pp.37-46
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• 2013

Objectives : The Meridians and acupuncture points are the fundamental theories for acupuncture therapy. They have been associated with nervous system, but It is not well defined. We investigated that the effects of acupuncture at the river points(LU8, HT4, PC5, SP5, KI7, LR4) on the changes in the expression of nNOS, iNOS, eNOS, and NE in rats. Methods : The Male Sprague-Dawley rats were divided into 6 groups each non-acupuncture and acupuncture group. We inserted needle and retained for 5 minutes on both left and right sides of LU8, HT4, PC5, SP5, KI7, LR4 which were the river points of five transport points for 6 yin meridian vessels. After that, blood was drawn via cardiac puncture, and tissues for each point near meridian vessels were extracted to examine the changes in the changes of nNOS, eNOS, iNOS and NE. Results : The LU8 and HT4 group showed a significant decrease on nNOS. In terms of eNOS and iNOS, the LU8 group decreased significantly while the KI7 group increased significantly. However, the experimental groups didn't show any significant changes on the plasma and tissue norepinephrine without plama NE in SP5 group. Conclusions : The effect on the nNOS, iNOS, eNOS of acupuncture at LU8 and KI7 could be observed, and it is considered that the effect of acupuncture related with on nervous system could be studied by additional researches based on this one.

• Korean Journal of Acupuncture
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• v.30 no.2
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• pp.97-103
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• 2013

Objectives : To observe the changes in the expression of nNOS, iNOS, eNOS and NO by the needle insertion on river points, one of the five transport points of three yang meridians of the foot. Methods : Based on rats, needle was inserted on both left and right sides of ST41, BL60 and GB38 and retained for five minutes. After the retention, blood was drawn via cardiac puncture and tissues from each point around meridian vessels were extracted to observe the changes in the expression of nNOS, iNOS, eNOS and NO. Results : In terms of the effect on expression of nNOS within tissues, ST41 showed significant decrease based on the results of immunohistochemistry. In terms of the effect on expression of iNOS within tissues, none of the experimental groups showed any significant change compared to the Normal group. Regarding expression of eNOS within tissues, GB38 showed significant increase based on the results of immunohistochemistry. In terms of the effect on NO creation in tissues, none of the experimental groups showed any significant change compared to the Normal group. Conclusions : The effect of needle insertion on the river points of three yang meridians of the foot on the function of nNOS, iNOS, eNOS and NO could be observed, and based on this study, it is considered that the effect of needle stimulation on the changes of nervous system could be found out through additional research.

• Advances in nano research
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• v.12 no.5
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• pp.501-514
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• 2022

This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

• Journal of Korean Neurosurgical Society
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• v.29 no.7
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• pp.877-885
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• 2000

Objective : The NOS inhibitors exhibit antinociceptive activity in rat model of neuropathic pain. NOS activity increases in the dorsal root ganglia(DRG) in neurop-athic pain. However, NOS activity decreases in the dorsal horn of spinal cord in the nerve injury models of neuropathic pain. To investigate whether the mechanism of decrease of NOS expression in the dorsal horn is related to a secondary effect resulting from increased NO production and likewise in the spinal DRG in the spinal nerve ligation model of neuropathic pain. Methods : We conducted behavioral tests for neuropathic pain, and nNOS immunohistochemistry and NADPH-diaphorase histochemistry after tight ligation of the 5th lumbar(L5) and 6th lumbar(L6) spinal nerves and L5 dorsal rhizotomy. Results : Typical neuropathic pain behaviors occurred 7 days after post-ligation in the neuropathic surgery group, but neuropathic pain behaviors in the dorsal rhizotomy group were absent or weak 7 days after post-operation. There was a decrease in the number of nNOS immunoreactive dorsal horn neurons on the both side(especially ipsilateral side) 7 days after post-ligation. The number of nNOS immunoreactive neurons in both side of the dorsal horn was not decreased 7 days after L5 dorsal rhizotomy. Conclusion : These data indicate that the changes in the injured DRG is essential for development and maintenance of neuropathic pain, and mechanism of decrease of nNOS expression in the dorsal horn is a secondary effect against the changes in the DRG including increased NO production in the spinal nerve ligation model of neuropathic pain.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.1
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• pp.28-35
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• 2008

Purpose: The nitric oxide (NO) release by inducible nitric oxide synthase (iNOS) is the key events in macrophage response to lipopolysaccharide (LPS) which is suggested to be a crucial mediator for inflammatory and innate immune responses. NO is an important mediator involved in many host defense action and may also lead to a harmful host response to bacterial infection. However, given the importance of iNOS in a variety of pathophysiological conditions, control of its expression and signaling events in response to LPS has been the subject of considerable investigation. Materials and Methods: The Raw264.7 macrophage cell line was used to observe LPS-stimulated iNOS expression. The expression of iNOS is observed by Western blot analysis and real-time RT-PCR. Protein kinase C $(PKC)-{\alpha}$ overexpressing Raw264.7 cells are established to determine the involvement of $PKC-{\alpha}$ in LPS-mediated iNOS expression. $NF-{\kappa}B$ activity is measured by $I{\kappa}B{\alpha}$ degradation and $NF-{\kappa}B$ luciferase activity assay. Results: We found that various PKC isozymes regulate LPS-induced iNOS expression at the transcriptional and translational levels. The involvement of $PKC-{\alpha}$ in LPS-mediated iNOS induction was further confirmed by increased iNOS expression in $PKC-{\alpha}$ overexpressing cells. $NF-{\kappa}B$ dependent transactivation by LPS was observed and $PKC-{\alpha}$ specific inhibitory peptide abolished this activation, indicating that $NF-{\kappa}B$ activation is dependent on $PKC-{\alpha}$. Conclusion: Our data suggests that $PKC-{\alpha}$ is involved in LPS-mediated iNOS expression and that its downstream target is $NF-{\kappa}B$. Although $PKC-{\alpha}$ is a crucial mediator in the iNOS regulation, other PKC isozymes may contribute LPS-stimulated iNOS expression. This finding is needed to be elucidated in further study.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.248.3-249
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• 2002

Nitric oxide (NO) has been shown to play an important role in the regulation of vascular tone. platelet function. neurotransmission. and immune function. NO is synthesized from the L-arginine by NO synthase (NOS). Three distinct isoforms of NOS have been identified: calcium/calmodulin-dependent endothelial (eNOS) and neuronal (nNOS) isoforms which are constitutive and produce small quantities of NO, and an inducible isoform (iNOS) which is markedly induced in response to lipopolysaccharide (LPS) or inflammatory cytokines. (omitted)

• The korean journal of orthodontics
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• v.40 no.4
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• pp.239-249
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• 2010

Objective: The aim of this study was to identify the expression of nitric oxide synthases (NOS) in the mandibular condyle during mandible advancement by functional appliance and to correlate it with the histologic changes and bone remodeling. Methods: Twenty-four female, 35-day-old Sprague-Dawley rats were randomly divided into 3 experimental groups. In all experimental groups, the mandibles of the rats were kept in a continuous forward position with a fixed bite jumping appliance. The rats were sacrificed on the 3rd, 14th, and 30th days of experiment. More than 2 rats in each group were used for staining. Results: There were no remarkable histologic changes and NOS expression differences in the control group. The most prominent histologic changes occurred in the 14th day experimental group. NOS decreased in the 30th day experimental group. There was increased expression of $NOS_2$ and $NOS_3$ in all experimental groups, comparative to the control group. In all the experimental groups and control group, the expression of $NOS_2$ was greater than that of $NOS_3$. Conclusions: It is postulated that $NOS_2$ and $NOS_3$ in the mandibular condyle might play an important role in bone remodelling of the mandibular condyle.