S-亚硝基-N-乙酰半胱氨酸对绿脓菌素感染鼠体内的病理形态及氧化作用的影响
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摘要
目的探讨S-亚硝基-N-乙酰半胱氨酸(SNAC)对绿脓菌素(PCN)感染鼠体内的病理形态及氧化作用的影响。方法 90只健康雄性SD大鼠采用随机数字表法分为3组,每组30只。对照组和PCN组大鼠分别腹腔注射1 ml/100 g 0.9%氯化钠溶液,SNAC组大鼠腹腔注射SNAC(1 ml/100 g)。用1%戊巴比妥钠溶液(40 mg/kg)行大鼠腹腔注射麻醉,对照组气管内接种0.9%氯化钠溶液0.2 ml,PCN组和SNAC组分别气管内接种PCN 0.2 ml(6×108CFU/ml)。分别于6、18、30、48、72 h观察大鼠一般状态,处死大鼠,取支气管肺泡灌洗液(BALF)和肺组织,观察肺组织形态学改变,计算BALF细胞总数和中性粒细胞计数,检测活性氧(ROS)含量、超氧化物歧化酶(SOD)活性、白介素8(IL-8)含量。结果 PCN组鼠感染6 h后出现精神萎靡、食欲不振、寒颤、发热、呼吸困难等,感染30 h时最明显;肺组织明显充血、水肿,大量炎性细胞浸润。大鼠PCN感染后6、18、30、48、72 h BALF细胞总数、BALF中性粒细胞计数、ROS含量、SOD活性、IL-8含量比较,差异均有统计学意义(P<0.05);其中PCN组和SNAC组各时间点BALF细胞总数、BALF中性粒细胞计数、ROS含量、IL-8含量较对照组均增高,SOD活性较对照组均降低,SNAC组各时间点BALF细胞总数、BALF中性粒细胞计数、ROS含量、IL-8含量较PCN组均降低,SOD活性较PCN组均增高(P<0.05)。结论 SNAC对PCN感染鼠体内氧化损伤具有一定的保护作用。
Objective To investigate the oxidative damage of mice infected by pyocyanin and its mechanism,and to further discuss the influence of S-nitroso-N-acetylcysteine( SNAC) on mice after infection. Methods 90 SD rats were randomly divided into three groups with each group 30 rats. The control group and PCN group were both given intraperitoneal injection of 0. 9% sodium chloride by 1 ml /100 g,while the SNAC group was given intraperitoneal injection of SNAC by 1 ml /100 g. The rats were also given intraperitoneal injection of 1% pentobarbital sodium by 40 mg / kg. The control group was inoculated with 0. 9% sodium chloride in the trachea,while the PCN group and SNAC group were inoculated with 0. 2 ml PCN( 6 × 108CFU / ml). The rats were killed at 6 h,18 h,30 h,48 h and 72 h in order to obtain the bronchoalveolar lavage fluid( BALF) and lung tissues. The general condition and morphology of lung tissues were observed,and the total number of BALF cells and neutrophil count were calculated. The ROS,SOD and IL-8 were also detected. Results After infection,the PCN group showed depression,loss of appetite,chills,fever and dyspnea at 6 h,and they aggravated at 30 h. The lung also had obvious hyperemia, edema and an infiltration of a large number of inflammatory cells. At 6 h,18 h,30 h,48 h and 72 h after infection,the total number of BALF cells,BALF neutrophil count,content of ROS,SOD activity and IL-8 showed statistically significant differences( P < 0. 05). Among them,the total number of BALF cells,BALF neutrophil count,content of ROS and IL-8 in PCN group and SNAC group at each time point were higher than those of the control group,while the SOD activity in PCN group and SNAC group at each time point was lower than that of the control group. The total number of BALF cells,BALF neutrophil count, content of ROS and IL-8 in the SNAC group at each time point were lower than those of the PCN group,while the SOD activity in the SNAC group at each time point was higher than that of the PCN group( P < 0. 05). Conclusion SNAC has protective effect on the oxidative damage of mice infected by pyocyanin.
Objective To investigate the oxidative damage of mice infected by pyocyanin and its mechanism,and to further discuss the influence of S-nitroso-N-acetylcysteine( SNAC) on mice after infection. Methods 90 SD rats were randomly divided into three groups with each group 30 rats. The control group and PCN group were both given intraperitoneal injection of 0. 9% sodium chloride by 1 ml /100 g,while the SNAC group was given intraperitoneal injection of SNAC by 1 ml /100 g. The rats were also given intraperitoneal injection of 1% pentobarbital sodium by 40 mg / kg. The control group was inoculated with 0. 9% sodium chloride in the trachea,while the PCN group and SNAC group were inoculated with 0. 2 ml PCN( 6 × 108CFU / ml). The rats were killed at 6 h,18 h,30 h,48 h and 72 h in order to obtain the bronchoalveolar lavage fluid( BALF) and lung tissues. The general condition and morphology of lung tissues were observed,and the total number of BALF cells and neutrophil count were calculated. The ROS,SOD and IL-8 were also detected. Results After infection,the PCN group showed depression,loss of appetite,chills,fever and dyspnea at 6 h,and they aggravated at 30 h. The lung also had obvious hyperemia, edema and an infiltration of a large number of inflammatory cells. At 6 h,18 h,30 h,48 h and 72 h after infection,the total number of BALF cells,BALF neutrophil count,content of ROS,SOD activity and IL-8 showed statistically significant differences( P < 0. 05). Among them,the total number of BALF cells,BALF neutrophil count,content of ROS and IL-8 in PCN group and SNAC group at each time point were higher than those of the control group,while the SOD activity in PCN group and SNAC group at each time point was lower than that of the control group. The total number of BALF cells,BALF neutrophil count, content of ROS and IL-8 in the SNAC group at each time point were lower than those of the PCN group,while the SOD activity in the SNAC group at each time point was higher than that of the PCN group( P < 0. 05). Conclusion SNAC has protective effect on the oxidative damage of mice infected by pyocyanin.
引文
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5 Husain M,Bourret TJ,McCollister BD,et al.Nitric oxide evokes an adaptive response to oxidative stress by arresting respiration[J].J Biol Chem,2008,283(12):7682-7689.
6 Andraus W,Souza GF,Oliveira MG,et al.S-nitroso-N-acetylcysteine ameliorates ischemia-reperfusion injury in the steatotic liver[J].Clinics,2010,65(7):715-721.
7 Foster MW,Hess DT,Stamler JS.Protein S-nitrosylation in health and disease:current perspective[J].Trends in Molecular Medicine,2009,15(9):391-404.
8 Han G,Zippin JH,Friedman A.From bench to bedside:the therapeutic potential of nitric oxide in dermatology[J].J Drugs Dermatol,2009,8(6):586-594.
9 Gusarov I,Shatalin K,Starodubtseva M,et al.Endogenous nitric oxide protects bacteria against a wide spectrum of antibiotics[J].Science,2009,325(5946):1380-1384.
10 Akaike T,Okamoto T,Zaki MH,et al.New paradigm of host defense against intracellular pathogens by nitric oxide[J].Nihon Hansenbyo Gakkai Zasshi,2009,78(1):41-47.
11 Keyer K,Imlay JA.Superoxide accelerates DNA damage by elevating free-iron levels[J].Proc Natl Acad Sci U S A,1996,93(24):13635-13640.
12 Schapiro JM,Libby SJ,Fang FC.Inhibition of bacterial DNA replication by zinc mobilization during nitrosative stress[J].Proc Natl Acad Sci U S A,2003,100(14):8496-8501.
13 Webert KE,Vanderzwan J,Duggan M,et al.Effects of inhaled nitric oxide in a rat model of Pseudomonas aeruginosa pneumonia[J].Crit Care Med,2000,28(7):2397-2405.
14 Rada B,Jendrysik MA,Pang L,et al.Pyocyanin-enhanced neutrophil extracellular trap formation requires the NADPH oxidase[J].PLoS One,2013,8(1):e54205.
15 罗招凡,劳伟思,梁伟南,等.血清同型半胱氨酸和SOD水平与急性脑梗死的相关性研究[J].中国卫生检验杂志,2011,21(10):2463-2464.
16 高天.颅内复杂动脉瘤患者围术期血清基质金属蛋白酶-9、超氧化物歧化酶、丙二醛及一氧化氮水平变化研究[J].实用心脑肺血管病杂志,2012,20(1):52.
17 苏显明,智晓文,刘景委,等.丹红注射液对老年冠心病患者血中对氧磷酶1、超氧化物歧化酶活性及丙二醛水平的影响研究[J].中国全科医学,2011,14(11):3804.
18 Kunkel SL,Standiford T,Kasahara K,et al.Interleukin-8(IL-8):the major neutrophil chemotactic factor in the lung[J].Exp Lung Res,1991,17(1):17-23.
19 程刚,陆士奇,李岩.急性胰腺炎患者血清降钙素原和肿瘤坏死因子α及白介素8的动态变化及临床意义[J].中国全科医学,2012,15(9):3180.
20 Zhou J,Jang YP,Kim SR,et al.Complement activation by photooxidation products of A2E,a lipofuscin constituent of the retinal pigment epithelium[J].Proc Natl Acad Sci U S A,2006,103(44):16182-16187.
21 Ito K,Ito M,Elliott WM,et al.Decreased histone deacetylase activity in chronic obstructive pulmonary disease[J].N Engl J Med,2005,352(19):1967-1976.
22 Gilmour PS,Rahman I,Donaldson K,et al.Histone acetylation regulates epithelial IL-8 release mediated by oxidative stress from environmental particles[J].Am J Physiol Lung Cell Mol Physiol,2003,284 (3):L533-540.
23 Ito K,Hanazawa T,Tomita K,et al.Oxidative stress reduces histone deacetylase 2 activity and enhances IL-8 gene expression:role of tyrosine nitration[J].Biochem Biophys Res Commun,2004,315(1):240-245.
24 Ivison SM,Wang C,Himmel ME,et al.Oxidative stress enhances IL-8 and inhibits CCL20 production from intestinal epithelial cells in response to bacterial flagellin[J].Am J Physiol Gastrointest Liver Physiol,2010,299(3):G733-741.
2 Muller M.Pyocyanin induces oxidative stress in human endothelial cells and modulates the glutathione redox cycle[J].Free Radic Biol Med,2002,33(11):1527-1533.
3 O'Malley YQ,Abdalla MY,McCormick ML,et al.Subcellular localization of Pseudomonas pyocyanin cytotoxicity in human lung epithelial cells[J].Am J Physiol Lung Cell Mol Physiol,2003,284(2):L420-430.
4 Fang FC.Antimicrobial reactive oxygen and nitrogen species:concepts and controversies[J].Nat Rev Microbiol,2004,2(10):820-832.
5 Husain M,Bourret TJ,McCollister BD,et al.Nitric oxide evokes an adaptive response to oxidative stress by arresting respiration[J].J Biol Chem,2008,283(12):7682-7689.
6 Andraus W,Souza GF,Oliveira MG,et al.S-nitroso-N-acetylcysteine ameliorates ischemia-reperfusion injury in the steatotic liver[J].Clinics,2010,65(7):715-721.
7 Foster MW,Hess DT,Stamler JS.Protein S-nitrosylation in health and disease:current perspective[J].Trends in Molecular Medicine,2009,15(9):391-404.
8 Han G,Zippin JH,Friedman A.From bench to bedside:the therapeutic potential of nitric oxide in dermatology[J].J Drugs Dermatol,2009,8(6):586-594.
9 Gusarov I,Shatalin K,Starodubtseva M,et al.Endogenous nitric oxide protects bacteria against a wide spectrum of antibiotics[J].Science,2009,325(5946):1380-1384.
10 Akaike T,Okamoto T,Zaki MH,et al.New paradigm of host defense against intracellular pathogens by nitric oxide[J].Nihon Hansenbyo Gakkai Zasshi,2009,78(1):41-47.
11 Keyer K,Imlay JA.Superoxide accelerates DNA damage by elevating free-iron levels[J].Proc Natl Acad Sci U S A,1996,93(24):13635-13640.
12 Schapiro JM,Libby SJ,Fang FC.Inhibition of bacterial DNA replication by zinc mobilization during nitrosative stress[J].Proc Natl Acad Sci U S A,2003,100(14):8496-8501.
13 Webert KE,Vanderzwan J,Duggan M,et al.Effects of inhaled nitric oxide in a rat model of Pseudomonas aeruginosa pneumonia[J].Crit Care Med,2000,28(7):2397-2405.
14 Rada B,Jendrysik MA,Pang L,et al.Pyocyanin-enhanced neutrophil extracellular trap formation requires the NADPH oxidase[J].PLoS One,2013,8(1):e54205.
15 罗招凡,劳伟思,梁伟南,等.血清同型半胱氨酸和SOD水平与急性脑梗死的相关性研究[J].中国卫生检验杂志,2011,21(10):2463-2464.
16 高天.颅内复杂动脉瘤患者围术期血清基质金属蛋白酶-9、超氧化物歧化酶、丙二醛及一氧化氮水平变化研究[J].实用心脑肺血管病杂志,2012,20(1):52.
17 苏显明,智晓文,刘景委,等.丹红注射液对老年冠心病患者血中对氧磷酶1、超氧化物歧化酶活性及丙二醛水平的影响研究[J].中国全科医学,2011,14(11):3804.
18 Kunkel SL,Standiford T,Kasahara K,et al.Interleukin-8(IL-8):the major neutrophil chemotactic factor in the lung[J].Exp Lung Res,1991,17(1):17-23.
19 程刚,陆士奇,李岩.急性胰腺炎患者血清降钙素原和肿瘤坏死因子α及白介素8的动态变化及临床意义[J].中国全科医学,2012,15(9):3180.
20 Zhou J,Jang YP,Kim SR,et al.Complement activation by photooxidation products of A2E,a lipofuscin constituent of the retinal pigment epithelium[J].Proc Natl Acad Sci U S A,2006,103(44):16182-16187.
21 Ito K,Ito M,Elliott WM,et al.Decreased histone deacetylase activity in chronic obstructive pulmonary disease[J].N Engl J Med,2005,352(19):1967-1976.
22 Gilmour PS,Rahman I,Donaldson K,et al.Histone acetylation regulates epithelial IL-8 release mediated by oxidative stress from environmental particles[J].Am J Physiol Lung Cell Mol Physiol,2003,284 (3):L533-540.
23 Ito K,Hanazawa T,Tomita K,et al.Oxidative stress reduces histone deacetylase 2 activity and enhances IL-8 gene expression:role of tyrosine nitration[J].Biochem Biophys Res Commun,2004,315(1):240-245.
24 Ivison SM,Wang C,Himmel ME,et al.Oxidative stress enhances IL-8 and inhibits CCL20 production from intestinal epithelial cells in response to bacterial flagellin[J].Am J Physiol Gastrointest Liver Physiol,2010,299(3):G733-741.