Does Plasma Nitric Oxide Concentration Correlate with the Pathological Grade of Rejection after Lung Allograft ?

Park, Young-Sik 梨花女子大學校 醫科大學 醫科學硏究所 1996 EMJ (Ewha medical journal) Vol.19 No.4

본 연구는 황견에서 동종 폐이식을 시행한 후에 합병된 거부반응시의 혈중 Nitric oxide농도와 병리조직학적 소견과의 상관관계를 보기위하여 시행되었다. 황견에서 일측 동종 폐 이식을 시행한 뒤, 면역억제제를 투여하였다. 실험견을 두군으로 분리하여 제1군(대조군 ; 황견 4마리)은 정상적으로 면역억제제를 투여하였다. 제2군(거부반응군 ; 황견 15마리)은 수술후 5일 부터 면역억제제의 투여를 중단하여 급성 거부반응을 유도하였다. 혈중 Nitric oxide는 수술전과 수술후 9일에 각군에서 각각 측정하여 비교하였다. 수술후 9일에 실험견을 희생시켜 병리조직학적 검사를 시행하여 급성 거부반응의 등급을 Grade 0-4로 분류하였다. 제2군에서 수술후 9일의 혈중 Nitric oxide농도와 거부반응의 병리 조직학적 등급과의 상관관계를 검사하였다. 제2군(거부반응군)에서 수술후 9일의 혈중 Nitricoxide농도가 수술전에 비하여 의의있게 증가되었다(12.28±2.10 vs 6.46±0.57uM/L; p<0.05). 그러나 제2군(거부반응군)에서 수술후 9일의 Nitric oxide농도와 거부반응의 병리조직학적 등급과의 상관관계는 유의하지 않았다.(상관계수 r=-0.2094 ; p >0.05). Objective : Experiments were designed to investigate whether there is any correlation between concentration of plasma nitric oxide and pathological severity of acute rejection after lung allograft. Methods : After single lung allograft, dogs were immunosuppressed with triple standard therapy and divided into 2 groups. Group 1(Immunosuppression ; n=4) was maintained on immunosuppression as controls. In group 2(Rejection ; n=15), triple therapy was discontinued to induce acute rejection from postoperative day 5. Plasma concentration of nitric oxide was measured by chemiluminescence method prior to surgery and at postoperative day 9. At postoperative day 9, dogs were sacrificed and rejection was graded pathologically by a working formulation for the standardization of nomenclature in the diagnosis of heart and lung rejection: lung rejection study group. Plasma nitric oxide level of day 9 was compared t that of prior to surgery in each group. Correlation between plasma nitric oxide level and pathological grade of acute rejection at day 9 in group 2(Rejection) was examined. Results : In group 2(Rejection), plasma nitric oxide level fo day 9 was elevated significantly, compared to that of prior to surgery(12.28±2.10 vs 6.46 0.57uM/L ; p<0.05).But plasma nitric oxide level of day 9 didn't significantly correlate with the pathological grades of rejection in group 2(Spearman's coefficient r=-0.2094 ; p>0.05). Conclusion : Plasma concentration of nitric oxide was elevated during acute rejection, but didn't correlate with the pathological severity of rejection.

산화질수(Nitric Oxide) 중단 후의 반동성폐고혈압

이현우,이재웅,현성열,박철현,박국양,이경천 대한흉부심장혈관외과학회 2000 Journal of Chest Surgery (J Chest Surg) Vol.33 No.2

Background: Inhaled nitric oxide therapy causes selective pulmonary vasodilation in congenital heart diseases with pulmonary hypertension. However discontinuation of inhaled nitric oxide therapy may be complicated by abrupt life-threatening rebound pulmonary hypertension(RPH) The purpose of this study was to prevent by comparing group I(without RPH n=13) and group II(with RPH n=6) to determine the risk factors involved inthe development of the RPH. Material and Method: Between Januarty 6, 1998 and April 14, 1999. we studied 19 consecutive children who were treated with inhaled nitric oxide for clinically significant pulmonary hypertension after an open heart surgery for congenital heart disease. the ratio of males and females was 12:7 ranging in age from 10 days to 6040 days(16 years) To identify the effects of nitric oxide between two groups we measured heart rate mean and systolic pulmonary arterial pressure mean and systolic systemic arterial pressure central venous pressure pH paO2/FiO2 and O2 saturation before and after the initiation and just before the withdrawal of the inhaled nitric oxide. result: In 6 of 19 patients(32%) withdrawal of inhaled nitric oxide caused RPH. In the two groups inhaled nitrix oxide decreased in pulmonary arterial pressure(PAP) without decreasing the systemic arterial pressure(SAP) and increased PaO2/FiO2 Compared with patients who had no RPH(group I) patients who had RPH(group II) were older in age (1204$\pm$1688 versus 546$\pm$1654 days p<0.05) received less nitric oxide therapy(34$\pm$18 versus 67$\pm$46 hours p<0.05) has shorter weaning process(5$\pm$3 versus 15一13 hours p<0.05) and received lowerconcentration of initial nitric oxide supply(11$\pm$8 versus 17$\pm$8 ppm p>0.05) and lower concentration just before the withdrawal nitric oxide(4.2$\pm$2.6 versus 5.6$\pm$2.6 ppm, p>0.05) Conclusion : We speculate that older age shorter of nitric oxide therapy shorter weaning process are the risk factors of RPH.

백서 패혈증 모델에서 HSP70의 과도 발현이 iNOS의 발현에 미치는 효과에 관한 연구

이용근,안융,임대호,백진아,고승오,신효근,Lee, Yong-Keun,Ahn, Yung,Leem, Dae-Ho,Baek, Jin-A,Ko, Seung-O,Shin, Hyo-Keun 대한구강악안면외과학회 2010 대한구강악안면외과학회지 Vol.36 No.5

Introduction: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. Materials and Methods: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. Results: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours ($47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, P=0.025) and 24 hours ($47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours ($5,513.73{\pm}1,051.60$ vs. $4,167.17{\pm}951.59$, P=0.025) and 24 hours ($18,740.27{\pm}8,241.20$ vs. $9,437.65{\pm}2,521.07$, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours ($0.86{\pm}0.48$ vs. $3.82{\pm}2.53$, P=0.016) and 24 hours ($0.39{\pm}0.25$ vs. $1.85{\pm}1.70$, P=0.025). Conclusion: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.

위선세포의 항산화 방어기전으로의 Nitric Oxide의 역할

김혜영(Hye-young Kim),이은주(Eun-joo Lee),김경환(Kyung Hwan Kim) 대한약리학회 1996 대한약리학잡지 Vol.32 No.3

위점막은 위강내에서 생성되는 독성이 강한 활성산소종에 노출된다. Nitric oxide는 glutathione의 항상성을 유지시킴으로써 acetaminophen 유도 간독성에 대한 보호효과를 나타내었다. 본 연구는 hydrogen peroxide로 인한 위선세포 손상에 대한 nitric oxide의 작용을 규명하고자 하였다. Hydrogen peroxide는 β-D-glucose와 glucose oxidase의 반응에 의해 생성시켰으며, 위선세포에 L-arginine, N^G-nitro-L-arginine methyl ester 및 N^G-nitro-L-arginine을 전처리 한 후, 세포외로 유리되는 지질과산화물 및 nitrite를 정량하고 세포내 glutathione 함량을 측정하였다. 결과로서, glucose/glucose oxidase를 처리한 경우 glucose oxidase 농도의존적으로 지질과산화물 생성은 증가되었으며, nitrite 유리 및 glutathione 함량은 감소되었다. NO synthase의 기질인 L-arginine 전처리시 glucose/glucose oxidase에 의한 지질과산화 및 nitrite 유리 증가와 세포내 glutathione 감소등이 방지되었다. N^G-nitro-L-arginine methyl ester 및 N^G-nitro-L-arginine등 NO synthase 억제제들은 세포손상에 보호효과를 나타내지 않았다. 결론적으로 nitric oxide는 hydrogen peroxide로 인한 세포손상에 대한 보호효과가 없으며, 이는 지질과산화 반응 및 세포내 glutathione 고갈등을 억제시킴으로써 이루어진다고 사료된다. Gatric mucosa is exposed to toxic, reactive oxygen species generated within the lumen. Nitric oxide protected acetaminophen-induced hepatotoxicity by maintaining glutathione homeostasis. The present study examined the role of nitric oxide in mediating hydrogen peroxide - induced damage to gastric cells. Hydrogen peroxide was generated by glucose oxidase acting on β-D-glucose. L-arginine, N^G-nitro-L-arginine methyl ester, or N^G-nitro-L-arginine were treated to the cells with glucose/glucose oxidase. Lipid peroxidation and nitrite release and cellular content of glutathione were determined. As a result, dose - dependent increase in lipid peroxide production as well as dose - dependent decrease in nitrite release and cellular glutathione content were observed in glucose/glucose oxidase - treated cells. Pretreatment of L-arginine, a substrate for nitric oxide synthase, prevented the increase of lipid peroxide production and the reduction of nitrite release as well as glutathione content. Inhibitors of nitric oxide synthase such as N^G-nitro-L-arginine methyl ester and N^G-nitro-L-arginine did not protect hydrogen peroxide - induced cell damage. In conclusion, nitric oxide protects gestric cells from hydrogen peroxide possibly by inhibiting lipid peroxidation and by preserving cellular glutathione stores.

Menadione에 의한 흰쥐 혈소판 세포독성에서 nitric oxide의 역할

승상애,김대병,윤여표,정진호 한국독성학회 1995 Toxicological Research Vol.11 No.2

Nitric oxide, a physiological transmitter, is reported to mediate cellular injury in various tissues. Its reactivity to free radical is believed to be one of the reasons for its involvement in cytotoxicity. Menadione, a representative quinone, is cytotoxic to several cell systems including isolated hepatocyte, endothelial cell and red blood cells. Its toxic mechanism is related to oxidative stress, mediated by toxic free radicals. Our previous studies demonstrated that menadione induced cell lysis and increase of oxygen consumption in platelets. It has been reported that platelets have nitric oxide producing enzyme, nitric oxide synthase. Thus, we have investigated to manifest the role of nitric oxide.in menadione-induced cytotoxicity in rat platelets. Menadione induced cytotoxicity in platelets was unaffected by $N^G$-nitro-arginine methyl ester (L-NAME), selective and competitive inhibitor of nitric oxide synthase. We also invesitgated the role of extracellular nitric oxide in menadione-induced cytotoxicity of platelets by addition with sodium nitroprusside (SNP). SNP did not affect platelet cytotoxicity by menadione. These results suggested that nitric oxide which was generated endogeneously or exogeneously might have a negligible role in menadione-induced cytotoxicity in rat platelets.

Role of Nitric Oxide as an Antioxidant in the Defense of Gastric Cells

김혜영,이은주,김경환,Kim, Hye-Young,Lee, Eun-Joo,Kim, Kyung-Hwan The Korean Society of Pharmacology 1996 대한약리학잡지 Vol.32 No.3

위점막은 위강내에서 생성되는 독성이 강한 활성산소종에 노출된다. Nitric oxide는 glutathione의 항상성을 유지시킴으로써 acetaminophen 유도 간독성에 대한 보호효과를 나타내었다. 본 연구는 hydrogen peroxide로 인한 위선세포 손상에 대한 nitric oxide의 작용을 규명하고자 하였다. Hydrogen peroxide는 ${\beta}-D-glucose$와 glucose oxidase의 반응에 의해 생성시켰으며, 위선세포에 L-arginine, $N^{G}-nitro-L-arginine$ methyl ester 및 $N^G-nitro-L-arginine$을 전처리 한 후, 세포외로 유리되는 지질과산화물 및 nitrite를 정량하고 세포내 glutathione 함량을 측정하였다. 결과로서, glucose/glucose oxidase를 처리한 경우 glucose oxidase 농도의존적으로 지질과산화물 생성은 증가되었으며, nitrite 유리 및 glutathione 함량은 감소되었다. NO synthase의 기질인 L-arginine 전처리시 glucose/glucose oxidase에 의한 지질과산화 및 nitrite 유리 증가와 세포내 glutathione 감소등이 방지되었다. $N^G-nitro-L-arginine$ methyl ester 및 $N^G-nitro-L-arginine$등 NO synthase 억제제들은 세포손상에 보호효과를 나타내지 않았다. 결론적으로 nitric oxide는 hydrogen peroxide로 인한 세포손상에 대한 보호효과가 없으며, 이는 지질과산화 반응 및 세포내 glutathione 고갈등을 억제시킴으로써 이루어진다고 사료된다. Gatric mucosa is exposed to toxic, reactive oxygen species generated within the lumen. Nitric oxide protected acetaminophen-induced hepatotoxicity by maintaining glutathione homeostasis. The present study examined the role of nitric oxide in mediating hydrogen peroxide - induced damage to gastric cells. Hydrogen peroxide was generated by glucose oxidase acting on ${\beta}-D-glucose$. L-arginine, $N^G-nitro-L-arginine$ methyl ester, or $N^G-nitro-L-arginine$ were treated to the cells with glucose/glucose oxidase. Lipid peroxidation and nitrite release and cellular content of glutathione were determined. As a result, dose - dependent increase in lipid peroxide production as well as dose - dependent decrease in nitrite release and cellular glutathione content were observed in glucose/glucose oxidase - treated cells. Pretreatment of L-arginine, a substrate for nitric oxide synthase, prevented the increase of lipid peroxide production and the reduction of nitrite release as well as glutathione content. Inhibitors of nitric oxide synthase such as $N^G-nitro-L-arginine$ methyl ester and $N^G-nitro-L-arginine$ did not protect hydrogen peroxide - induced cell damage. In conclusion, nitric oxide protects gestric cells from hydrogen peroxide possibly by inhibiting lipid peroxidation and by preserving cellular glutathione stores.

패혈증에서 Heat Shock Protein 70의 과도발현이 Inducible Nitric Oxide Synthase의 발현에 미치는 효과

이용근 ( Yong Keun Lee ),신효근 ( Hyo Keun Shin ),권운용 ( Woon Yong Kwon ),서길준 ( Gil Joon Suh ),윤여규 ( Yeo Kyu Youn ) 대한외상학회 2008 大韓外傷學會誌 Vol.21 No.1

Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: 47.19 ± 10.04 vs. 33.22 ± 8.28, p = 0.025) and 24 h (CLP+GLN vs. CLP: 47.06 ± 10.60 vs. 31.90 ± 4.83, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: 4167.17 ± 951.59 vs. 5513.73 ± 1051.60, p = 0.025) and 24 h (CLP+GLN vs. CLP: 9,437.65 ± 2,521.07 vs. 18,740.27 ± 8,241.20, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: 0.86 ± 0.48 vs. 3.82 ± 2.53 umol/L, p = 0.016) and 24 h (CLP+GLN vs. CLP: 0.39 ± 0.25 vs. 1.85 ± 1.70 umol/L, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

Nitric oxide-releasing polymer incorporated ointment for cutaneous wound healing

Kang, Y.,Kim, J.,Lee, Y.M.,Im, S.,Park, H.,Kim, W.J. Elsevier Science Publishers 2015 Journal of controlled release Vol.220 No.2

This work demonstrates the development of nitric oxide-releasing ointment and its potential on efficient wound healing. Nitric oxide-releasing polymer was successfully synthesized, which is composed of biocompatible Pluronic F127, branched polyethylenimine and 1-substituted diazen-1-ium-1,2-diolates. The synthesized nitric oxide-releasing polymer was incorporated into the PEG-based ointment which not only facilitated nitric oxide release in a slow manner, but also served as a moisturizer to enhance the wound healing. As compared to control groups, the nitric oxide-releasing ointment showed the accelerated wound closure with enhanced re-epithelialization, collagen deposition, and blood vessel formation in vivo. Therefore, this nitric oxide-based ointment presents the promising potential for the efficient strategy to heal the cutaneous wound.

Nitric oxide induced by Indian ginseng root extract inhibits Infectious Bursal Disease virus in chicken embryo fibroblasts in vitro

( Bhaskar Ganguly ),( Vijaypillai Umapathi ),( Sunil Kumar Rastogi ) 한국축산학회(구 한국동물자원과학회) 2018 한국축산학회지 Vol.60 No.1

Infectious Bursal Disease is a severe viral disease of chicken responsible for serious economic losses to poultry farmers. The causative agent, Infectious Bursal Disease virus, is inhibited by nitric oxide. Root extract of the Indian ginseng, Withania somnifera, inhibits Infectious Bursal Disease virus in vitro. Also, Withania somnifera root extract is known to induce nitric oxide production in vitro. Therefore, the present study was undertaken to determine if the inhibitory activity of Withania somnifera against Infectious Bursal Disease virus was based on the production of nitric oxide. We show that besides other mechanisms, the inhibition of Infectious Bursal Disease virus by Withania somnifera involves the production of nitric oxide. Our results also highlight the paradoxical role of nitric oxide in the pathogenesis of Infectious Bursal Disease.

Analysis of Blood Flow-dependent Blood Nitric Oxide Level and Half-life of Nitric Oxide in Vivo

Kim Cuk-Seong,Kim Hyo-Shin,Lee Young-Jun,Park Jin Bory,Ryoo Sung-Woo,Chang Seok-Jang,Jeon Byeong-Hwa Biomedical Engineering Society for Circulation 2003 International Journal of Vascular Biomedical Engin Vol.1 No.2

Endothelial release of nitric oxide (NO) contributes to the regulation of vascular tone by inducing vascular relaxation. To estimate the blood flow-dependent nitric oxide level and half-life (T1/2) of nitric oxide in vivo state, we investigated the change of aortic NO currents during the change of aortic blood flow rate using NO-selective electrode system and electromagnetic flowmeter in the aorta of anesthetized rats. Resting mean aortic blood flow rate was $49.6{\pm}5.6ml/min$ in the anesthetized rats. NO currents in the aorta were increased by the elevation of blood pressure and/or blood flow rate. When the aortic blood flow was occluded by the clamping, aortic NO currents were decreased. The difference of NO concentration between resting state and occluded state was $1.34{\pm}0.26{\mu}M$ (n=7). This NO concentration was estimated as blood flow-dependent nitric oxide concentration in the rats. Also, while the aortic blood flow was occluded, NO currents were decreased with exponential pattern with $12.84{\pm}2.15$ seconds of time constant and $7.70{\pm}1.07$ seconds of half-life. To summarize, this study suggested that blood flow-dependent NO concentration and half-life of nitric oxide were about $1.3{\mu}M$ and 7.7 seconds, respectively, in the aorta of anesthetized rats. The nitric oxide-selective electrode system is useful for the direct and continuous measurement of NO in vivo state.